Immunomodulatory Effects and Mechanisms of Curcuma Species and Their Bioactive Compounds: A Review

Abstract

Curcuma species (family: Zingiberaceae) are widely utilized in traditional medicine to treat diverse immune-related disorders. There have been many scientific studies on their immunomodulating effects to support their ethnopharmacological uses. In this review, the efficacy of six Curcuma species, namely, C. longa L., C. zanthorrhiza Roxb., C. mangga Valeton & Zijp, C. aeruginosa Roxb. C. zedoaria (Christm.) Roscoe, and C. amada Roxb., and their bioactive metabolites to modulate the immune system, their mechanistic effects, and their potential to be developed into effective and safe immunomodulatory agents are highlighted. Literature search has been carried out extensively to gather significant findings on immunomodulating activities of these plants. The immunomodulatory effects of Curcuma species were critically analyzed, and future research strategies and appropriate perspectives on the plants as source of new immunomodulators were discussed. Most of the pharmacological investigations to evaluate their immunomodulatory effects were in vivo and in vitro experiments on the crude extracts of the plants. The extracts were not chemically characterized or standardized. Of all the Curcuma species investigated, the immunomodulatory effects of C. longa were the most studied. Most of the bioactive metabolites responsible for the immunomodulating activities were not determined, and mechanistic studies to understand the underlying mechanisms were scanty. There are limited clinical studies to confirm their efficacy in human. Of all the bioactive metabolites, only curcumin is undergoing extensive clinical trials based on its anti-inflammatory properties and main use as an adjuvant for the treatment of cancer. More in-depth studies to understand the underlying mechanisms using experimental in vivo animal models of immune-related disorders and elaborate bioavailability, preclinical pharmacokinetics, and toxicity studies are required before clinical trials can be pursued for development into immunomodulatory agents.

Full text

Immunomodulatory Effects and
Mechanisms of Curcuma Species and
Their Bioactive Compounds: A Review
Yuandani
1
*, Ibrahim Jantan
2
, Ade Sri Rohani
1
and Imam Bagus Sumantri
3
1
Department of Pharmacology, Faculty of Pharmacy, Universitas Sumatera Utara, Medan, Indonesia,
2
Institute of Systems
Biology, Universiti Kebangsaan Malaysia, Selangor, Malaysia,
3
Department of Pharmaceutical Biology, Faculty of Pharmacy,
Universitas Sumatera Utara, Medan, Indonesia
Curcuma species (family: Zingiberaceae) are widely utilized in traditional medicine to treat
diverse immune-related disorders. There have been many scientific studies on their
immunomodulating effects to support their ethnopharmacological uses. In this review,
the efficacy of six Curcuma species, namely, C. longa L., C. zanthorrhiza Roxb.,C. mangga
Valeton & Zijp, C. aeruginosa Roxb. C. zedoaria (Christm.) Roscoe, and C. amada Roxb.,
and their bioactive metabolites to modulate the immune system, their mechanistic effects,
and their potential to be developed into effective and safe immunomodulatory agents are
highlighted. Literature search has been carried out extensively to gather significant findings
on immunomodulating activities of these plants. The immunomodulatory effects of
Curcuma species were critically analyzed, and future research strategies and
appropriate perspectives on the plants as source of new immunomodulators were
discussed. Most of the pharmacological investigations to evaluate their
immunomodulatory effects were in vivo and in vitro experiments on the crude extracts
of the plants. The extracts were not chemically characterized or standardized. Of all the
Curcuma species investigated, the immunomodulatory effects of C. longa were the most
studied. Most of the bioactive metabolites responsible for the immunomodulating activities
were not determined, and mechanistic studies to understand the underlying mechanisms
were scanty. There are limited clinical studies to confirm their efficacy in human. Of all the
bioactive metabolites, only curcumin is undergoing extensive clinical trials based on its anti-
inflammatory properties and main use as an adjuvant for the treatment of cancer. More in-
depth studies to understand the underlying mechanisms using experimental in vivo animal
models of immune-related disorders and elaborate bioavailability, preclinical
pharmacokinetics, and toxicity studies are required before clinical trials can be pursued
for development into immunomodulatory agents.
Keywords: curcuma species, ethnopharmacology, phytochemicals, immunomodulation, immune system
INTRODUCTION
The human body has a remarkably sophisticated immune system consisting of white blood cells and
specialized immune molecules that protect the body against invading pathogens (Tan and Vanitha,
2004). The immune system is made up of innate and adaptive immune immunity. Innate immunity
provides first protection against pathogens, and then it will stimulate adaptive immunity to enhance
Edited by:
Ipek Suntar,
Gazi University, Turkey
Reviewed by:
Md. Areeful Haque,
International Islamic University
Chittagong, Bangladesh
Hari Prasad Devkota,
Kumamoto University, Japan
*Correspondence:
Yuandani
yuandani@usu.ac.id
yuan_dani@yahoo.com
Specialty section:
This article was submitted to
Ethnopharmacology,
a section of the journal
Frontiers in Pharmacology
Received: 17 December 2020
Accepted: 18 February 2021
Published: 30 April 2021
Citation:
Yuandani, Jantan I, Rohani AS and
Sumantri IB (2021) Immunomodulatory
Effects and Mechanisms of Curcuma
Species and Their Bioactive
Compounds: A Review.
Front. Pharmacol. 12:643119.
doi: 10.3389/fphar.2021.643119
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431191
REVIEW
published: 30 April 2021
doi: 10.3389/fphar.2021.643119

the protection. Innate immunity is the most rapidly acting
immunity. It mostly depends on neutrophils, macrophages,
dendritic cells, and monocytes, while T and B cells are
involved in adaptive immunity (Beutler, 2004;Saroj et al.,
2012). In response to pathogens, leukocytes perform a number
of phagocytic activities, including chemotaxis, leukocytes
adhesion to vascular endothelial cells, and pathogen
engulfment, followed by intracellular killing to eliminate the
pathogens (Beutler, 2004;Kobayashi et al., 2005). Phagocytes
migrate toward the chemoattractants such as complement (C3a
and C3b) and formyl methionyl-leucyl-phenylalanine (fMLP) (a
bacterial product) (Luster, 2001). Chemoattractants utilize a
similar signal transduction system, namely, G protein–coupled
receptor, that is, platelet-activating factor receptor (PAFR),
formyl-methionyl-leucyl-phenylalanine receptor (fMLPR), and
complement C5a receptor (C5aR). The interaction of chemotactic
factor and its receptor stimulates cytoskeletal reorganization,
calcium mobilization, and degranulation in heterologous cell
types (Firtel and Chung, 2000). The adhesion of leukocytes to
vascular endothelial cells is initiated by selection interaction,
followed by the interaction of leukocyte integrin of the CD18
complex on the surface of phagocytes with adhesion molecule on
endothelial cells (Beutler, 2004). Phagocytosis of microorganism
triggers superoxide radical (O
2-
) generation and release of
reactive oxygen species (ROS) such as hydroxyl radical,
hypochlorous acid (HOCl), and chloramines through the
activity of myeloperoxidase (MPO). Besides, macrophages are
involved in the release of nitric oxide (NO
.
) by inducible nitric
oxide synthase (iNOS) (Bogdan, 2001).
Macrophages also modulate adaptive immunity by presenting
antigen to CD4 T cells through major histocompatibility complex
(MHC) class II antigen. CD4 T cells perform their functions by
four subpopulations, which include Th-1, Th-2, Th-17, and
CD4 T regulatory (Treg) cells (Chapel et al., 2006). Th cells
help B cells develop into plasma cells which can produce antibody
and also activate T cells to become activated cytotoxic T cells
(Beutler, 2004;Luckheeram et al., 2011). Several cytokines also
play essential roles in immune response, which consist of pro-
inflammatory cytokines such as tumor necrosis factor-alpha
(TNF-α), interleukin 1 (IL-1), IL-6, IL-11, IL-8, and anti-
inflammatory cytokines or cytokines inhibitor such as IL-4,
IL-10, and IL-13. Cytokines as intercellular messenger
molecules have several functions, and these include stimulating
phagocyte migration and coordinating early responses of
monocytes, macrophages, dendritic cells, and lymphocytes
during inflammatory states (Shaikh, 2011). The release of pro-
inflammatory cytokines is regulated by nuclear factor-kappa B
(NF-ĸB) and mitogen-activated protein kinase (MAPK) pathways
(Beyaert et al., 2013). Defects or malfunctions in the immune
system can cause disorders of the immune system. Inappropriate
reaction to self-antigen is known as autoimmunity such as
myasthenia gravis, type 1 diabetes (T1D), systemic lupus
erythematosus, Graves’disease, celiac disease, pernicious
anemia, rheumatoid arthritis, and multiple sclerosis.
Overactive immune response is known as hypersensitivity
reactions, while ineffective immune response is known as
immunodeficiency (Zhernakova et al., 2009;Warrington et al.,
2011;Beyaert et al., 2013). The diseases which cause the body’s
immune system to attack the small intestine has affected 1 in 133
people in the United States (Rattue, 2012). A review on incidence
and prevalence of Crohn’s disease in several countries reported a
gradual increase in incidence and prevalence of this disease. In
Malaysia, a study during 2001–2003 showed an increase of
prevalence especially among Indians, compared to Chinese
and Malay populations. Meanwhile, in Singapore, a study
showed that majority patients were Chinese, and there was a
trend of increased of prevalence (Economou et al., 2009).
Therefore, modulation of the immune response is required in
the management and treatment of diseases due to immune system
dysfunction (Geetha et al., 2005).
The treatment of inflammatory and immune-related diseases
due to defects or disorders of the immune system necessitates
modulation of the immune response. Immunomodulation is the
process of modifying an immune response by administration of a
drug or compound, while immunomodulators are substances
which are used to modulate the components of the immune
system (Patil et al., 2012). There are several chemical
immunomodulators available in the market, that is,
prednisone, hydrocortisone, and dexamethasone, which have
been used to treat numerous inflammatory diseases.
Recombinant proteins have emerged as one important drug to
treat cancer, immunodeficiency, and infectious diseases.
Cyclosporin A, a microbial peptide, is the most widely used
immunosuppressant in transplant rejection treatment (Elgert,
2009). Unfortunately, most of these commercial drugs have
side effects. Gastric and intestinal mucosal damage are the
commonest adverse effects of NSAIDS. Corticosteroids, an
immunosuppressive drug, show various side effects, such as
reduced bone marrow and increased skin fragility. Cyclosporin
A exhibited toxicities and side effects including nephrotoxic
activity and gingival hyperthrophy. Therefore, safer and more
effective drugs are required as alternatives. Natural products
remain one of the important sources of new and safe anti-
inflammatory agents (Elgert, 2009).
In an effort to investigate for safer drugs,
ethnopharmacological information can be used to provide
preliminary data in the search for new drugs. It can be an
indicator of pharmacological activity of natural products that
could be further investigated for their mechanisms of action in
cellular, animal, and human studies (Flores, 2017). Among them,
some therapeutic activities of plant extracts or compounds have
been proposed to be due to their effects on the immune system.
Many plants of the genus Curcuma, especially C. longa,C.
zanthorrhiza,C. amada,C. mangga,C. aeruginosa, and C.
zedoaria, were reported to modulate the immune functions
and possessed a variety of immunomodulatory effects. The
strong immunomodulatory activity of these plants was due to
their bioactive compounds as their main constituents.
Curcuminoids, particularly curcumin, have been reported as
the major components of plants in Curcuma species. Besides,
other compounds, such as xanthorrhizol, have been reported to
be present in other Curcuma species. A number of reviews on the
phytochemistry, and biological and pharmacological activities of
the genus Curcuma have been published recently (Rajkumari and
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431192
Yuandani et al. Immunomodulatory Effects of Curcuma Species

Sanatombi, 2017;Sun et al., 2017;Dosoki and Setzer, 2018;
Chanda and Ramachandra, 2019;Kaliyadasa and
Samarasinghe, 2019;Kavitha and Mahadevi, 2020;USDA,
2021). However, there is either no or little and unconcise
reports on the immunomodulatory effects of genus Curcuma
and their bioactive molecules in these articles. In this review, we
elaborated the ability of C. longa L.,C. zanthorrhiza Roxb.,C.
mangga Valeton & Zijp, C. aeruginosa Roxb.,C. zedoaria
(Christm.) Roscoe, and C. amada Roxb.and their bioactive
metabolites to modulate the immune response in different
lineages of the immune system.
METHODS
This comprehensive review was based on updated scientific
databases on six major Curcuma species, namely, C. longa,C.
zanthorrhiza,C. manga,C. aeruginosa,C. zedoaria, and C.
amada. Databases were scanned from January 2000 until
December 2020 for animal, in vitro, and clinical studies. A
systematic search of databases with the use of the keywords
“curcuma AND immune system,”“curcumin AND immune
system,”and each species of Curcuma genus, such as
“Curcuma mangga AND immune system,”“Curcuma longa
AND immune system,”was carried out. Only published data
were included in this study; meanwhile, references without title in
English were excluded. Literature search has been carried out
extensively to gather data, involving use of published scientific
reports in Frontiers, the Science Direct, Scopus, Google Scholar,
the Institute for Scientific Information (ISI)-Web of Science, Pub
Med, Wiley Online Library, Elsevier, Springer, Taylor and
Francis, ACS Publications Today, and other references over
the past two decades. The gathered data on the
immunomodulating effects of the Curcuma species were
critically analyzed, and future strategies and appropriate
perspectives for the plants as a source of new natural
immunomodulators were discussed.
TAXONOMY AND DISTRIBUTION
Curcuma L.is one of the largest genera in the family of
Zingiberaceae, and there are approximately 100 accepted
Curcuma species. It is found throughout tropical Asia from
India to South China, Southeast Asia, Papua New Guinea, and
northern Australia (Dosoky and Setzer, 2018). The word
“curcuma”is derived from the Arabic word “kurkum,”which
means yellow color (Kaliyadasa and Samarasinghe, 2019).
Curcuma species are originated from the Indo-Malayan region
and widespread in Asia, Africa, and Australia (Sasikumar, 2005).
Figure 1 shows the Curcuma species: C. longa,C. zanthorrhiza,C.
amada,C. mangga,C. aeruginosa, and C. zedoaria that are
discussed in this review. The rhizomes of these plants are
widely utilized in traditional medicine and as spices, food
flavors, colorants, cosmetics, and perfumery. C. longa Linn
(syn. Curcuma domestica Val.) is native to tropical South Asia,
but it has been found throughout tropical areas (Li et al., 2011),
such as Cambodia, China, India, Nepal, Indonesia, Madagascar,
Malaysia, the Philippines, and Vietnam (Yadav and Tarun, 2017).
It is commonly called as turmeric (Li et al., 2011;HMPC, 2017;
Rajkumari and Sanatombi, 2017) and the Golden Spice of India
(Yadav and Tarun, 2017). C. longa has been associated to the
Indian culture for nearly 4000 years and probably reached China
by 700 AD, East Africa by 800 AD, and West Africa by 1200 AD
(Yadav and Tarun, 2017). C. longa has a specific name in some
regions, namely, Haridra (Sanskrit, Ayurvedic), Jianghuang
(Chinese), Kyoo or Ukon (Japanese) (HMPC, 2017), kurkum
(Arabic), and haldi (Hindi and Urdu) (Dosoky and Setzer, 2018).
C. longa has yellow-white flowers, 10–15 cm of stalk length, the
seeds are brown ovoid, the plant grows upright, and part used for
FIGURE 1 | Plants and rhizomes of Curcuma species. (A) Cucuma zanthorrhiza,(B) Curcuma mangga,(C) Curcuma longa,(D) Curcuma amada (Artfire, 2016;
Snapdeal, 2020), (E) Curcuma zedoaria, and (F) Curcuma aeruginosa.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431193
Yuandani et al. Immunomodulatory Effects of Curcuma Species

spices and medicine is the rhizome (Tung et al., 2019). C.
zanthorrhiza is native to Indonesia (Rajkumari and Sanatombi,
2017), and it has been established by the Food and Drug
Supervisory Agency (BADAN POM) as one of the leading
medicinal plants (Ervintari et al., 2019). It is known as temu
lawak (Dewi et al., 2012) and Java turmeric (Kim M-B et al., 2014;
Astana et al., 2018), and distributed in Southeast Asia. It has been
grown in Thailand, the Philippines, Sri Lanka, and Malaysia (Oon
et al., 2015). It is grown simply to produce rhizomes which are
commonly used in folk medicine (Wahono et al., 2017b). It is an
ethnomedicinal plant from Indonesia and Malaysia (Kim M-B
et al., 2014). It has 2-m tall erect pseudostems (Rajkumari and
Sanatombi, 2017) and is generally cultivated in village home
gardens. The rhizomes smell balmy and taste bitter (Ilene et al.,
2020). C. zanthorrhiza has been used as an active ingredient in
cosmetic and hygienic products in Germany and the Netherlands
(Drugbank, 2021).
C. amada is widely distributed in Myanmar, and in southern
and eastern India. Apart from Myanmar, C. amada is also
distributed in the tropics of Asia to Africa and Australia. It is
widely cultivated in West Bengal, Gujarat, Tamil Nadu, and the
northeastern states of India (Sasikumar, 2005). It has the
resemblance with ginger (Zingiber officinale) but imparts a raw
mango (Mangifera indica)flavor (Policegoudra et al., 2011).
Thereby, it is usually known as mango ginger due to its
mango flavor. The flavor has been attributed to the presence
of cis-ocimene and car-3-ene (Ayodele et al., 2018). C. amada
rhizomes are fleshy, buff colored, 5–10 cm long, and 2–5cm in
diameter (Artfire, 2016;Policegoudra et al., 2011;Snapdeal,
2020). C. aeruginosa is an endemic species in Myanmar, but it
is also distributed in West Bengal and Kerala (Rajkumari and
Sanatombi, 2017). C. aeruginosa is also an ethnomedicinal plant
in Indonesia, Malaysia, Thailand, Northern Australia, and Papua
New Guinea (Sulfianti et al., 2019). It is commonly known as Kali
Haldi (in India) and has a deep-blue or bluish-black colored
cortex with pungent odor. In Indonesia, C. aeruginosa is known
as Temu Ireng (Choudhury et al., 2013;George and Britto, 2015),
and in English, it is known as pink and blue ginger (Sulfianti et al.,
2019), temu hitam in Malaysia, and waan-maha-mek in Thailand.
It is a perennial herb derived from Burma and spread to tropical
countries in Malaysia, Thailand, India, and Indonesia (Dosoky
and Setzer, 2018). C. zedoaria, known as white turmeric, is a
perennial herb with perpendicular pseudostem and fleshy roots.
It is a native plant from Bangladesh, India, and Sri Lanka (Lobo
et al., 2009), but it is a critically threatened species in Bangladesh
and India (Anisuzzaman et al., 2008). It is known by several
names in India, and the most common are Krachura (Sanskrit),
Gandamatsi (Hindi), and Sutha (Bengali) (Lobo et al., 2009). In
China, it is generally called Ezhu (Lee et al., 2019). C. zedoaria is
widely cultivated in subtropical regions (Southeast Asia,
Thailand, Indonesia, Japan, and China). From outside, C.
zedoaria looks like ginger, but inside, it looks like turmeric
(Dosoky and Setzer, 2018). C. zedoaria rhizome has dark
orange-fleshed tubers (Rahayu et al., 2020). C. mangga
rhizome is commonly known as mango turmeric as it has the
mango-like smell as in C. amada. It is a perennial herb with
30–110 cm of stem height. It is native from Java (Rajkumari and
Sanatombi, 2017). It is distributed in most tropical countries such
as Indonesia, Thailand, and Malaysia (Hong et al., 2016).
ETHNOPHARMACOLOGICAL USES
C. longa is traditionally used as an antioxidant, anti-
inflammatory, antidiabetic, hepatoprotective, and
anticarcinogenic agent (Alsahli et al., 2018). It is well known
as ethnomedicinal plant and used in different traditional systems
in the world. In traditional medicine in Nigeria, C. longa is also
used as an wound-healing agent (Adeshina et al., 2017). In Nepal,
C. longa is applied as an anthelmintic, a tonic and blood purifier
as well as for the treatment of Jaundice and liver disorder. In Peru,
C. longa juice commonly known as Shapi natiyu is applied for the
treatment of bronchitis and malaria. In Colombia, it is used for
circulatory stimulant, healing wounds, liver cleaning, immune
system booster, thrombosis, indigestion, diabetes, high
cholesterol, and kidney infection (Ayati et al., 2019). The
Ayurvedic Pharmacopoeia of India documented that C. longa
is used as tonic, stomachic, and carminative. In Chinese
Pharmacopoeia, C. longa has a potential for eliminating blood
stasis, stimulating menstruation discharge, and relieving pain
(Yue et al., 2010). In Pakistan traditional medicine, C. longa is
used as a wound-healing agent and for the treatment of pimples.
In Butanese folk medicine, it is known as Yung-ba and applied as
tonic, antidote, antiseptic, anti-inflammatory, and as a good
preservative (Ayati et al., 2019). C. zanthorrhiza is traditionally
used for wound healing, as anti-inflammatory and
anticarcinogecic agent, and for lowering of serum cholesterol
levels (Kim et al., 2007) and booster immunity by Javanese
(Setyati et al., 2019). In Malaysia, it is traditionally used to
treat skin inflammation, rheumatism, stomach and liver
disorders, and hepatitis (Kim M-B et al., 2014). In Ayurveda,
C. amada is usually used for inflammation, asthma, bronchitis,
biliousness, and skin disease (Policegoudra et al., 2006). The
rhizomes are usually used for anorexia, dyspepsia, chronic ulcers,
pruritus, gout, and inflammations (Thokchom and Phucho,
2015). Traditionally, C. amada is used for inflammation,
stomach and skin diseases, cough, and rheumatism in
Myanmar (Win et al., 2017). C. aeruginosa is used to booster
immunity by Javanese (Setyati et al., 2019). It is used traditionally
in Indonesia for gastrointestinal disease, and as antimicrobial and
anti-inflammatory agents (Sulfianti et al., 2019). C. zedoaria is
commonly known as white turmeric, and it is widely used as a
traditional medicine in Indonesia (Putri, 2014;Aristyani et al.,
2018), China and Japan (Kim et al., 2001), and India (Nan et al.,
2014). C. zedoaria is traditionally used for treating cancer (Dutta,
2015) and also used as a traditional remedy to promote blood
circulation in Korea and Japan (Kim et al., 2001). C. zedoaria is
used to treat flatulent colic, hepatocirrhosis, and cancer in
traditional Chinese medicine. It is also used to treat blood
stagnation syndromes and for promoting menstruation
(Carvalho et al., 2010). C. mangga is highly valued in
Indonesian folk medicine for its healing properties in the
treatment of stomach disorders, fever, and cancer-related
diseases (Malek et al., 2011).
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431194
Yuandani et al. Immunomodulatory Effects of Curcuma Species

PHYTOCHEMISTRY
Plants from the genus Curcuma L. have been intensively studied
for their phytochemical contents and bioactivity due to their
tremendous ethnopharmacological and therapeutic potentials.
There are recent reviews on the phytochemistry, and biological
and pharmacological activities of Curcuma species (Rajkumari
and Sanatombi, 2017;Dosoki and Setzer, 2018;Chanda and
Ramachandra, 2019;Kavitha and Mahadevi, 2020;USDA,
2021). Phytochemical analysis has revealed that Curcuma
species are made up mainly of terpenoids, flavonoids, phenolic
compounds, organic acids, anthocyanin, tannins, and inorganic
compounds. Until now, phytochemical studies on 32 Curcuma
species have isolated and identified a total of 719 compounds,
which include 529 terpenoids, 15 flavonoids, 102
diphenylalkanoids, 19 phenylpropene derivatives, 3 alkaloids, 7
steroids, and 44 other types of compounds (Sun et al., 2017). The
phytochemical content of C. longa has been extensively
investigated, and more than 235 compounds have been
identified in the rhizome, which are mainly polyphenols and
terpenoids. The major group of polyphenols is curcuminoids,
which may contain up to 80% of curcumin, and other two are
demethoxycurcumin and bisdemethoxycurcumin. In total, there
are 109 sesquiterpenes, 68 monoterpenes, 22 diarylheptanoids
and diarylpentanoids, eight phenylpropene and other phenolic
compounds, five diterpenes, four sterols, three triterpenoids, two
alkaloids, and 14 other compounds (Li et al., 2011). The essential
oils of flowers and leaves are mainly made up of monoterpenes,
while the root and rhizome oils are dominated by sesquiterpenes.
A recent study reported that the average essential oil content in
the rhizome was 3.97%, and the major components identified by
gas chromatography were ar-turmerone (40%), α-turmerone
(10%), and curlone (23%) (Guimarães et al., 2020).
Xanthorrhizol, a bisabolane-type sesquiterpenoid compound, is
the major compound of C. zanthorrhiza. Curcumin,
demethoxycurcumin, and bisdemethoxycurcumin are also
present in appreciable amounts. Sesquiterpenes of the
bisabolene-type and their oxygenated derivatives were reported
to comprise more than 92% of the rhizome oil of C. zanthorrhiza.
Xanthorrhizol (32%) was the most abundant sesquiterpene
phenol. ß-Curcumene (17.1%), zingiberene (13.2%), ß-
bisabolol (3.5%), and ar-curcumene (2.6%) were the other
major components of the oil (Jantan et al., 2012).
Several valuable sesquiterpenoids such as zedoarondiol
zedoalactone A, zedoalactone B, curcumenol, isocurcumenol,
zedoarol, isofuranodiene, and furanodiene have been isolated
from C. aeruginosa rhizome. The rhizome oil of this plant was
made up mainly of 1, 8-cineol, ß-pinene, camphor, curzerenone,
furanodienone, furangermenone, curcumenol, zedoarol,
isocurcumenol, and ß-elemene (Jose and Thomas, 2014). C.
zedoaria rhizome is rich in sesquiterpenoids which are
represented by furanodienone, furanodiene, curzerenone,
zedorone, germacrone, curzeone, 13-hydroxy germacrone,
curcumenol, curcumenone, dihydrocurdione, zedoaronediol,
dihydrocurdione, zedoarol, 13-hydroxygermacrone, curzeone
curcumenone, curcumanolide-A, curcumanolide-B,
a-turmerone, ß-turmerone, epicurzerenone, and curzerene. GC
and GC-MS analyses of the rhizome oil revealed the presence of
curzerenone (22.3%) as the major component, together with 1,8-
cineole, germacrone, cymene, a-phellandrene, and ß-eudesmol
(Lobo et al., 2009). Based on percent yield, myrcene (88.6%),
ocimene (47.2%), and ar-turmerone (29.12%) were reported to be
the major chemical constituents of C. amada. Other compounds
that were present in appreciable amounts were (Z)-β-farnesene,
guaia-6,9-diene, cis ß-ocimene, cis-hydroocimene, trans-
hydroocimene, a-longipinene, a-guaiene, linalool, ß-
curcumene, and turmerone (Jatoi et al., 2007).
The presence of these diverse bioactive compounds in the
plants contributes to the diverse pharmacological activities.
Curcumin, one of the main active ingredients in Curcuma
species, has been widely reported for its strong
immunomodulating, antioxidant, anti-inflammatory, and
antitumor activities. Structure–activity relationship studies
have revealed that the presence of different functional entities
on the diarylheptanoid structure which include methoxy,
phenoxy, and carbon–carbon double bonds was found to be
responsible for the antioxidant property. However, the
remarkable anti-inflammatory property was associated with
the symmetry of the structure and position of substituents
along with the number of methoxy groups. In addition,
electron-withdrawing substituents and the α,β-unsaturated
carbonyl group were indicated imperative for reactivity
(Arshad et al., 2017). Besides the curcuminoids (curcumin,
demethoxycurcumin, bisdemethoxycurcumin, and
dihydrocurcumin), other compounds from Curcuma spp. with
significant activity on the immune system include xanthorrhizol,
turmeronol, curdione, curcuzedoalide, curcumenol, and
germacrone.
IMMUNOMODULATING PROPERTIES OF
CURCUMA SPECIES
Curcuma species and their bioactive compounds have been much
investigated for their various biological and pharmacological
activities, including antioxidant, anti-inflammatory, anticancer,
hepato-protective, antifungal, antihypertensive, neuroprotective,
and immunomodulatory effects through in vitro and in vivo
studies. The six Curcuma species and their bioactive
compounds discussed in this article have been documented to
exhibit various pharmacological activities, particularly via
modulation of the immune system. There are in-depth
mechanistic studies on the immunomodulating effects of some
of these species available in the literature. The
immunomodulatory effects of the plant samples on the
immune system are critically analyzed, and their underlying
mechanisms of action are summarized in Table 1.
Curcuma longa L.
In Vitro Immunomodulating Effect of C. longa
Of all the Curcuma species investigated, the immunomodulatory
effects of C. longa were the most studied. Interestingly, most
experimental studies on the extracts of C. longa were carried out
using in vivo animal models, and there were few in vitro studies. The
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431195
Yuandani et al. Immunomodulatory Effects of Curcuma Species

TABLE 1 | Immunomodulatory activity of some Curcuma species.
Species Subjects Study
design
Preparation Immunomodulatory
activities
Modulation Parameters/
mediators affected
References
Curcuma
amada Roxb.
Rat PMNs in vitro Ethanol, petroleum
ether, chloroform, and
acetone extracts
Phagocytosis activity ↑Phagocytosis Karchuli and
Pradhan (2011)
Sheep RBC-induced
albino Wistar rats
in vivo Ethanol extract Cellular immunity ↑Delayed-type
hypersensitivity
response
Karchuli and
Pradhan (2011)
Sheep RBC induced-
albino Wistar rats
in vivo Ethanol extract Humoral immunity ↑Antibody titer Karchuli and
Pradhan (2011)
Curcuma
aeruginosa
Roxb.
Zymosan-stimulated
human PMNs
in vitro Methanol extract ROS generation ↓ROS Jantan et al.
(2011)
Zymosan-stimulated
macrophages of BALB/c
mice
in vitro Methanol extract ROS generation ↓ROS Jantan et al.
(2011)
Human PMNs in vitro Methanol extract PMN chemotaxis ↓Chemotaxis Jantan et al.
(2011)
Human whole blood in vitro Methanol extract CD18/11a expression ↓CD18/11a Harun et al.
(2015)
Human whole blood in vitro Methanol extract Phagocytosis activity ↓Phagocytosis Harun et al.
(2015)
Lymphocytes of BALB/c
mice
in vitro Extract by steam
distillation
Counts of CD4
+
and
CD8
+
cells
↑CD4
+
and CD8
+
cells Anggriani et al.
(2019)
DMBA-induced Wistar
rats
in vivo Ethanol extract Cytokine release ↑TNF-α, IFN-γ, IL-2,
and IL-12
Sulfianti et al.
(2019)
Epinephelus fuscoguttatus in vivo C. aeruginosa, Piper
retrofractum, and C.
zanthorrhiza water
extracts
Leukocyte number ↑Total leukocyte count Setyati et al.
(2019)
Epinephelus fuscoguttatus in vivo C. aeruginosa, Piper
retrofractum, and C.
zanthorrhiza water
extracts
Phagocytosis activity ↑Phagocytic index Setyati et al.
(2019)
Curcuma
longa Linn
CMS-induced
Sprague–Dawley rats
in vivo Ethanol extract Cytokine release ↓IL-6 and TNF-αXia et al. (2006)
Male Sprague–Dawley rats in vivo Ethanol extract Splenic NK cell activity ↑NK cell Xia et al. (2006)
Mice in vivo Methanol extract Adaptive immune
response
↑Leukocytes number,
antibody titer, spleen
index, and delayed-
type hypersensitivity
response
Kumolosasi et al.
(2018)
Human peripheral blood
mononuclear cells
(PBMCs)
in vitro Polar fraction of hot
water extract
Proliferation response ↑PBMC viability Yue et al. (2010)
Human peripheral blood
mononuclear cells
(PBMCs)
in vitro Polysaccharide-
enriched fraction at
200 μg/ml
Cytokine gene
expression
↑GM-CSF, IL-1, IL-5,
IL-8, IL-10, and IL-13
Yue et al. (2010)
Human peripheral blood
mononuclear cells
(PBMCs)
in vitro Polysaccharide-
enriched fraction at
400 and 800 μg/ml
Cytokine release ↑TNF-αand IL-6 Yue et al. (2010)
Human peripheral blood
mononuclear cells
(PBMCs)
in vitro Polysaccharide-
enriched fraction at
800 μg/ml
Cytokine release ↑TGF-βYue et al. (2010)
Human peripheral blood
mononuclear cells
(PBMCs)
in vitro Polysaccharide-
enriched fraction at
800 μg/ml
Lymphocyte population ↑CD14
+
Yue et al. (2010)
Unstimulated mouse
splenocytes and mouse
macrophage (RAW264.7)
cells
in vitro Water extract Cytokine release ↑NO, IL-2, IL-6, IL-10,
IL-12, IFN-γ, TNF-α,
and MCP-1
Chinampudur
et al. (2013)
LPS stimulated mouse
splenocytes
in vitro Water extract Cytokine release ↓NO, IL-12, IL-6, and
PGE
2
Chinampudur
et al. (2013)
Con-A–induced
splenocytes
in vitro Water extract Cytokine release ↑IL-2 and IFN-γChinampudur
et al. (2013)
(Continued on following page)
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431196
Yuandani et al. Immunomodulatory Effects of Curcuma Species

TABLE 1 | (Continued) Immunomodulatory activity of some Curcuma species.
Species Subjects Study
design
Preparation Immunomodulatory
activities
Modulation Parameters/
mediators affected
References
Con-A–induced
splenocytes
in vitro Water extract Cytokine release ↓IL-10 Chinampudur
et al. (2013)
LPS-unstimulated and
stimulated mouse
splenocytes
in vitro Polysaccharide
fraction
Lymphocyte proliferation ↑Splenocytes number Chinampudur
et al. (2013)
LPS-stimulated mouse
splenocytes
in vitro Polysaccharide
fraction
Cytokine release ↑IL-10 Chinampudur
et al. (2013)
LPS-stimulated mouse
splenocytes
in vitro Polysaccharide
fraction
Cytokine release ↓IL-12 and PGE
2
Chinampudur
et al. (2013)
RAW264.7 macrophages in vitro Water extract Nitric oxide (NO)
production
↑NO levels Pan et al. (2017)
Diabetic infected rats in vivo Ethanol extract Total IgE ↓IgE levels Shabana et al.
(2020)
Diabetic infected rats in vivo Ethanol extract Leukocyte number ↓Total leukocyte
count (TLC)
Shabana et al.
(2020)
Diabetic infected rats in vivo Ethanol extract NO production ↓NO Shabana et al.,
2020
Diabetic infected rats in vivo Ethanol extract Cytokine release ↓IL-6, TNF-a, and IL-1βShabana et al.,
2020
LP-BM5 MuLV-induced
mice
in vivo Alcohol extract Proliferation ↓T-cell, B-cell, and NK-
cell
Kim O. K. et al.
(2014)
LP-BM5 MuLV-induced
mice
in vivo Alcohol extract Cytokine imbalance Prevented Th1 (IL-2 and IFN-
γ)/Th2) (IL-4 and
IL-10)
Kim O. K. et al.
(2014)
C57BL/6 mice in vivo C. longa, Mulberry
leaves, and purple
sweet potato extracts
Proliferation ↓T cell and B cell Yoo et al. (2013)
C57BL/6 mice in vivo C. longa, Mulberry
leaves, and purple
sweet potato extracts
Cytokine secretion ↓Th 1 cytokines (IL-2
and IFN-γ), Th2
cytokines (TNF-αand
IL-10)
Yoo et al. (2013)
LP-BM5 MuLV-infected
mice
in vivo C. longa and sweet
potato mixture
Messenger RNA (mRNA)
expression
↑MHC I and MHC II Park et al. (2018)
LP-BM5 MuLV-infected
mice
in vivo C. longa and sweet
potato mixture
Population of CD4
(+)/CD8 (+) T cells
↓CD4 (+)/CD8 (+)
T cells
Park et al. (2018)
LP-BM5 MuLV-infected
mice
in vivo C. longa powder and
sweet potato mixture
Ig levels ↓IgA, IgE, and IgG Park et al. (2018)
Human umbilical vein
endothelial cells (HUVECs)
in vitro Extract mRNA levels ↓NF-κB p65, IL-6, and
TNF-α
Morales et al.
(2012)
C57BL mice in vivo Hot water extract Cytokines release ↓TNF-α, IL-6, and IL-6
m-RNA proteins
Uchio et al.
(2017)
Fusarium root in vivo Aqueous extract mRNA of the defense-
related genes
↑Defensin and
chitinase
Alsahli et al.
(2018)
Clarias gariepinus in vivo Powder IgM level ↑IgM Adeshina et al.
(2017)
Clarias gariepinus in vivo Powder Enzyme activity ↑Lysozyme activity Adeshina et al.
(2017)
Cyprinus carpio in vivo Powder Leukocyte number ↑Neutrophils,
lymphocytes,
monocyctes,
eosinophils, and
basophils
Arunkumar et al.
(2016)
Fish green terror
(Andinocara rivulatus)
in vivo Powder White blood cell number ↑White blood cells Mooraki et al.
(2019)
Nile tilapia (Oreochromis
niloticus)
in vivo Powder Leukocrit levels ↑Leukocrit number Hassan et al.
(2018)
M. rosenbergii in vivo Powder Gene expression ↑Crustin and lysozyme Alambra et al.
(2012)
Chicks in vivo Powder Lymphocyte percentage ↑Lymphocytes Naderi et al.
(2014)
Curcuma
zedoaria
Rosc.
LPS-stimulated
RAW264.7 cells
in vitro Methanol extract NO production ↓NO Lee et al. (2019)
(Continued on following page)
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431197
Yuandani et al. Immunomodulatory Effects of Curcuma Species

TABLE 1 | (Continued) Immunomodulatory activity of some Curcuma species.
Species Subjects Study
design
Preparation Immunomodulatory
activities
Modulation Parameters/
mediators affected
References
LPS-stimulated
RAW264.7 cells
in vitro Methanol extract Pro-inflammatory protein
expression
↓iNOS and COX-2 Lee et al. (2019)
RBL-2H3 cells in vitro Aqueous acetone
extract
Beta-hexosaminidase
release
↓Beta-hexosaminidase Lobo et al. (2009)
C57Bl/6J mice in vivo Ethanol extract Total leukocytes count ↑Leukocytes Carvalho et al.
(2010)
L. monocytogenes and S.
aureus–stimulated
RAW264.7 cells
in vitro Essential oil Cytokine release ↓TNF-αHuang et al.
(2019)
PMA-stimulated
RAW264.7 cells
in vitro Polysaccharide
fraction
Cytokine release ↑TNF-αKim et al. (2001)
PMA-stimulated
RAW264.7 cells
in vitro Polysaccharide
fraction
NO production ↑NO Kim et al. (2001)
Curcuma
zanthorrhiza
Roxb.
Zymosan-stimulated
human whole blood
in vitro Methanol extract ROS generation ↓ROS Jantan et al.
(2011)
Zymosan-stimulated
PMNs
in vitro Methanol extract ROS generation ↓ROS Jantan et al.
(2011)
Zymosan-stimulated
macrophages of BALB/c
mice
in vitro Methanol extract ROS generation ↓ROS Jantan et al.
(2011)
Human PMNs in vitro Methanol extract PMN chemotaxis ↓Chemotaxis Jantan et al.
(2011)
Human whole blood in vitro Methanol extract Expression of CD18/11a ↓CD18/11a Harun et al.
(2015)
Human whole blood in vitro Methanol extract Phagocytosis activity ↑Phagocytosis Harun et al.
(2015)
Hypercholesterolemic
male Sprague–Dawley rats
in vivo Curcuminoid cider IL1β, TNFα,and
chemokine gene
expression
↓IL1β,TNFα,and
chemokine
Hardiwati et al.
(2019)
High cholesterol diet male
Sprague–Dawley rats
in vivo Curcuminoid cider CD44, ICAM-1, iNOS,
and LOX-1 gene
expression
↓CD44, ICAM-1, iNOS,
and LOX-1
Mauren et al.
(2016)
Human lymphocytes in vitro Volatile oil Lymphocytes
proliferation
↑Lymphocytes Miksusanti (2012)
Alcohol-induced mice in vivo Ethanol extract Lymphocytes
proliferation
↓Lymphocytes Ilene et al. (2020)
High-fat diet-induced
C57BL/6 mice
in vivo Ethanol extract Cytokine genes
expression
↓TNF-α, IL-6, IL-1β,
and C-reactive
protein (CRP)
Kim M-B et al.
(2014)
RAW 264.7 cells in vitro Crude polysaccharide
extract
Chemical mediators
release
↑TNF-αand PGE
2
Kim et al. (2007)
RAW 264.7 cells in vitro Crude polysaccharide
extract
Oxidative burst ↑NO and H
2
O
2
Kim et al. (2007)
RAW 264.7 cells in vitro Crude polysaccharide
extract
Phosphorylation ↑IκBαKim et al. (2007)
LPS-stimulated human
gingival fibroblast-1 cells
in vitro Crude polysaccharide
extract
mRNA levels ↓IL-1β, NF-κB p65,
MMP-2, and MMP-8
Kim et al. (2018)
HIV/AIDS patients Clinical
study
C. zanthorrhiza in
combination with C.
mangga and
Phyllanthus niruri
Lymphocytes
proliferation
Maintained CD4
+
value Astana et al.
(2018)
Systemic lupus
erythematosus (SLE)
patients
Clinical
study
C. zanthorrhiza
supplementation with
vitamin D3
Cytokine release No significant
difference
reduction
IL-6 Wahono et al.
(2017a)
Systemic lupus
erythematosus (SLE)
patients
Clinical
study
C. zanthorrhiza
supplementation with
vitamin D3
Cytokine release No significant
difference
reduction
IL-17 Wahono et al.
(2017b)
Systemic lupus
erythematosus (SLE)
patients
Clinical
study
Powder Cytokine release ↓TNF-αSetiawati et al.
(2017)
(Continued on following page)
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431198
Yuandani et al. Immunomodulatory Effects of Curcuma Species

followings are reports on the few in vitro studies that have been
carried out to evaluate the immunomodulating effects of C. longa.C
longa fermented by Aspergillus oryzae (FCL) exhibited
immunomodulatory effects in RAW 264.7 cells. The different
extracts of FLC on phagocytic activity, TNF-α,NOproduction,
NK cell activity, and mRNA expression of LP-BM5 eco displayed the
following results: hot water and 20% ethanol extracts increased the
phagocytic activity, but there was no significant change in the
production of NO relative to the control. There was also
suppression of mRNA expression of LP-BM5 eco in FCL extracts
and a four-fold increase in NK cell cytotoxity relative to the control
group, especially in the 20% ethanol extract treatment group.
However, TNF-αwas significantly increased by the addition of
FCL extracts (Yoo et al., 2014). Curcuminoid extract from C.
longa has been reported to modulate TNF-⍺and IL-6 at protein
and gene levels in adipocytes in vitro (Hardiwati et al., 2019). C. longa
decreased mRNA levels of NF-κBp65,IL-6,andTNF-αat 2.5–5mg/
L in LPS-induced human umbilical vein endothelial cells (HUVEC)
(Morales et al., 2012). The polysaccharide extract isolated from C.
longa was reported to possess immunostimulatory activities.
Investigation of the polar fractions of C. longa hot water extract
displayed that the extract stimulated PBMC proliferation using the
[methyl-3H]-thymidine incorporation assay. Furthermore, its
polysaccharide-enriched fraction at 200 μg/ml enhanced the
cytokine expression (IL-1, IL-5, IL-8, IL-10, IL-13, and GM-CSF)
detected by semiquantitatively using the antibody-based RayBio
human cytokine array. However, the fraction at 200 μg/ml did
not significantly enhance TNF-α,IFN-γ,TGF-β,andIL-6
productions. The production of IL-6 and TNF-αwas only
enhanced after treatment with the fraction at the higher doses of
400 and 800 μg/ml, respectively. The polysaccharide fraction at
800 μg/ml stimulated TGF-βrelease and CD14+ lymphocyte and
TABLE 1 | (Continued) Immunomodulatory activity of some Curcuma species.
Species Subjects Study
design
Preparation Immunomodulatory
activities
Modulation Parameters/
mediators affected
References
Curcuma
mangga Val.
Swiss albino mice in vivo Ethanol extract and its
fraction (hexane,
chloroform, ethyl
acetate, and aqueous
fractions)
Paw and ear edema ↓Paw and ear volume Ruangsang et al.
(2010)
LPS and IFNγ–induced
RAW264.7 macrophage
cells
in vitro Methanol extract NO production ↓NO Abas et al. (2006)
LPS-stimulated
RAW264.7 macrophage
cells
in vitro Ethanol extract and
chloroform, hexane,
and ethyl acetate
fractions
NO production ↓NO Kaewkroek et al.
(2009)
Zymosan-stimulated
human whole blood
in vitro Methanol extract ROS inhibitory activity ↓ROS Jantan et al.
(2011)
Zymosan-stimulated
human PMNs
in vitro Methanol extract ROS inhibitory activity ↓ROS Jantan et al.
(2011)
Zymosan-stimulated
macrophages of BALB/c
mice
in vitro Methanol extract ROS inhibitory activity ↓ROS Jantan et al.
(2011)
Human PMNs in vitro Methanol extract PMN chemotaxis ↓Chemotaxis Jantan et al.
(2011)
Human whole blood in vitro Methanol extract Expression of CD18/11a ↓CD18/11a Harun et al.
(2015)
Human whole blood in vitro Methanol extract Phagocytosis activity ↑Phagocytosis Harun et al.
(2015)
Mice in vivo n-Hexane, ethyl
acetate, and ethanol
extracts
Phagocytosis activity ↑Phagocytosis Yuandani and
Suwarso
(2017b);
Yuandani et al.
(2019)
Bovine RBC-stimulated
mice
in vivo Ethanol extract Humoral immunity ↑Antibody titer Yuandani et al.
(2018)
Bovine RBC-stimulated
mice
in vivo Ethanol extract Cellular immunity ↑Delayed-type
hypersensitivity
response
Yuandani et al.
(2018)
Doxorubicin-induced
immunosuppressive rats
in vivo Ethanol extract Humoral immunity ↑Antibody titer Yuandani et al.
(2020)
Doxorubicin-induced
immunosuppressive rats
in vivo Ethanol extract Cellular immunity ↑Delayed-type
hypersensitivity
response
Yuandani et al.
(2020)
↑, increase.
↓, decrease.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 6431199
Yuandani et al. Immunomodulatory Effects of Curcuma Species

population. However, the CD4+/CD8+ ratio was not altered after
administration with polysaccharide fraction (Yue et al., 2010). In a
related study, the immunostimulant and anti-inflammatory effects
of C. longa aqueous extract and its polysaccharide fractions in the
presence and absence of mitogens were determined. The extract
enhanced splenocyte proliferation in unstimulated and LPS or
concanavalin A-stimulated cells. The extract increased the levels
ofIL-2,IL-10,NO,IL-6,IL-12,TNF-α,IFN-γ,andMCP-1inthe
absence of mitogen. Interestingly, C. longa extract decreased the
levels of IL-12, IL-6, NO, and PGE-2 in LPS-stimulated cells, while
TNF-α, IL-10, and MCP-1 levels were not altered. In contrast, the
extract stimulated IL-2 and IFN-γproduction but decreased IL-10
production from Con-A–induced splenocytes. Furthermore, its
polysaccharide fraction showed stimulatory activity on
lymphocyte proliferation in the absence or presence of LPS. The
levels of IL-10 were increased, but the levels of IL-12 and PGE-2 were
decreased after treatment with C. longa in LPS-stimulated cells
(Chinampudur et al., 2013). In another study, a C. longa root
aqueous extract standardized to a minimum of 20% of
polysaccharides ukonan A, B, C, and D was shown to stimulate
NO production in RAW264.7 macrophages (Pan et al., 2017).
In Vivo Immunomodulating Effect of C. longa
Most immunomodulating studies were carried out using aqueous
and alcoholic extracts. The ethanol extract of C. longa was
reported to suppress immune function, and behavioral and
neuroendocrine alterations in a rat chronic mild stress (CMS)
model. The enhancement of cytokine level (TNF-αand IL-6)
activity and NK cell activity inhibition in the CMS-induced rat in
splenocytes were reversed by administration of 35 mg/kg of C.
longa ethanol extract and 7 mg/kg of fluoxetine as a control. The
putative antidepressant properties of the extract were due to
suppressive effects on cytokine biosynthesis. However, the extract
increased the IL-6 level in the nonstress group, but there was no
significant difference as compared with those of the normal group
and caused a slight but no significant decrease in TNF-αlevels.
Although the extract enhanced splenic NK cell activity in CMS-
treated rats, the NK cell activity of nonstressed rat did not change
after treatment with C. longa (Xia et al., 2006). In another study,
treatment with C. longa methanol extract with a single dose of
200 mg/kg for 14 days in mice stimulated innate and adaptive
immunity. The effect of the extract on adaptive immunity was
investigated by immunizing and challenging the mice with sheep
red blood cells (sRBCs) on days 7 and 14, respectively. C. longa
enhanced the adaptive immunity by increasing leukocyte
number, antibody titer, spleen index, and delayed-type
hypersensitivity response (Kumolosasi et al., 2018). However,
the results of this study are preliminary as different doses of the
extract need to be used to determine a dose–response relationship
and the optimal dose for efficacy.
A previous study reported that treatment with C. longa in
diabetic rats infected with Staphylococcus aureus resulted in a
decrease of IgE, total leukocyte number (TLC), NO, and
cytokine production (IL-6, IL-1β,andTNF-α). The results
indicated that there was improvement of immune function
by reducing levels of pro-inflammatory cytokines in the
diabetic rats (Shabana et al., 2020). It was reported that 20%
C. longa alcohol extract suppressed the increase of liver weights,
lymph node, and spleen, and reduction of proliferation of T and
B cells and NK cell activity stimulated by murine leukemia
viruses–induced murine acquired immunodeficiency syndrome
(AIDS) infection. Moreover, the extract suppressed Th1/Th2
(IL-2, IFN-γ/IL-4, and IL-10) cytokine imbalance and pro-
inflammatory cytokine production (Kim O-K et al., 2014).
This is in agreement with another study which showed that a
diet consisted of C. longa; mulberry leaves and purple sweet
potato extracts have the ability to prevent splenomegaly and
lymphadenopathy induced by retrovirus, decrement of B- and
T-cell proliferation, as well as reduction of Th 1 cytokine (IFN-γ
and IL-2) release. It also reduced Th2 cytokine (TNF-αand IL-
10) release (Yoo et al., 2013). Moreover, C. longa alone and in
combination with purple sweet potato inhibited LP-BM5
murine leukemia virus (MuLV)-induced lymphadenopathy.
The mixture of C. longa and purple sweet potato at the doses
of 2 and 5 g/kg body weight increased the mRNA expression of
MHC I and II as compared to those of the infected control
group. The mixture at 5 g/kg body weight decreased the
population of CD4
+
T cells as compared to the infected
control group, and also, the population of CD8
+
T cells was
lower than that of the normal group. Moreover, the extracts also
affected T- and B-cell proliferation. The levels of Th1-type
cytokines (IL-12 and IL-15) were enhanced after treatment
bythemixture;meanwhile,Th2-typecytokine(IL-4,IL-10,
IL-6, and TNF-α) production was significantly decreased as
compared to the infected control group. In addition, the mixture
at the doses of 2 and 5 g/kg decreased the levels of IgA, IgE, and
IgG. Besides, C. longa alone or in mixture enhanced the
phagocytosis activity of LP-BM5 MuLV-infected mice (Park
et al., 2018). C. longa hot water extract protected the C57BL
mice liver from acute injury induced by ethanol at 3 g/kg. The
hepatic injury caused an increase in TNF-α, IL-6, and IL-6
m-RNA proteins. However, an increase in these proteins was
not found in mice treated with hot water extract of C. longa
30 min before induction (Uchio et al., 2017). C. longa aqueous
extract has been evaluated for its immunotherapeutic and
hepatoprotective activities in CCl
4
intoxicated Swiss albino
mice. The aqueous extract reduced the levels of bilirubin and
transaminase enzymes (SGOT and SGPT) in mice. Treatment
with CCl
4
resulted in liver damage and reduced nonspecific
host–response parameters such as NO and MPO release,
phagocytosis, intracellular killing capacity of peritoneal
macrophages, and morphological alteration. Treatment with
the extract also significantly protected the adverse effects of
CCl
4
on the nonspecific host response in the peritoneal
macrophages of the mice (Sengupta et al., 2011).
Interestingly, there are several studies on the ability of C. longa
to modulate the immune response of fish, chick, and prawn. C.
longa increased plant defense by enhancing the defense-related
genes such as defensin and chitinase of treated sunflower
seedlings (Alsahli et al., 2018). The enhancement of host
defense in fish has also been reported. C. longa leaf–enriched
diet was fed to the fish to satiation twice daily for 12 weeks. Then,
the fish was challenged with Aeromonas hydrophila. The highest
stimulation on immunoglobulin M (IgM) level and lysozyme
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311910
Yuandani et al. Immunomodulatory Effects of Curcuma Species

activity was observed in fish fed with 2.5% C. longa–fortified diets
(Adeshina et al., 2017). A study reported that Mesocyclops
thermocyclopoides enriched with C. longa enhanced the
differential leukocyte number in fish (Cyprinus carpio),
including enhancement of neutrophils, lymphocytes,
monocytes, eosinophils, and basophils (Arunkumar et al.,
2016). This result was supported by a previous study which
reported the ability of 0.3% turmeric powder–enriched fish
diet to enhance the white blood cell number significantly as
compared to those of the control group (Mooraki et al., 2019).
Turmeric in combination with rosemary (Rosmarinus officinalis)
and thyme (Thymus vulgaris) increased the leukocrit levels in fish
(Hassan et al., 2018).
C. longa was also able to enhance the immune response of
prawns (Macrobrachium rosenbergii) after being infected by
Vibrio alginolyticus. Identification using RT-PCR revealed that
C. longa–enhanced feeds increased the gene expression of crustin
and lysozyme in M. rosenbergii, indicating a remarkable increase
in the expression of AMPs (antimicrobial peptides). Production
of AMPs is a first-line host defense mechanism of innate
immunity, and they are thought to be essential for organisms
lacking adaptive immunity (Alambra et al., 2012). The ability of
C. longa to modulate the immune response in chicks was also
reported. C. longa powder constituted 2.5 and 7.5 g/kg of the diet,
which significantly enhanced lymphocyte percentage in chicks.
Supplementation of the diet with the powder at 2.5 g/kg of the
diet resulted in a significant increase in anti-infectious bronchitis
virus (IBV) titer compared to the control group (Naderi et al.,
2014). In another study, 2.5% of C. longa–enriched diet protected
chicken from Salmonella pullorum infection (Purwanti et al.,
2018). Moreover, the cellular immunity of broiler chicken to
phytohemagglutinin-P (PHA-P) was significantly higher in
groups fed with higher amount of C. longa. The primary
antibody titer to sRBCs was also stimulated (Sethy et al.,
2017). These studies revealed that C. longa mostly enhanced
the cellular and humoral responses of fish, chick, and prawns.
Thus, this plant can be used as animal feed to enhance the
immune defense of the animals.
Extensive cellular and animal studies have been performed to
evaluate the immunomodulatory effects of C. longa by using
various immune cells such as macrophages, monocytes,
neutrophils, lymphocytes (T and B cells), and NK cells. There
is a need to explore the immune effect of the plant with other
immune cells, particularly the antigen-presenting cells such as
dendritic cells. The existing reports should be supported by
exploring the effects of the plant samples on various animal
disease models of immune-related and chronic inflammatory
disorders. All the extracts of Curcuma species used in the
in vitro and in vivo immunomodulating studies were not
analyzed for their chemical constituents or standardized to
marker compounds. C. longa samples were mostly in the form
of crude aqueous and alcoholic extracts. Some of the samples
were curcuminoids or polysaccharide-rich extracts, but the
chemical composition of the extracts were not determined. It
has been suggested that the curcuminoids and polysaccharides
might be the main contributors for immunomodulatory activity
of the plant. The extracts used should be determined qualitatively
and quantitatively by using validated analytical methods such as
reversed-phase HPLC methods. Some of the bioactive
compounds—especially the curcuminoids—have been isolated
from the extracts, and their mechanistic effects in modulating the
immune system have been determined.
Curcuma zanthorrhiza Roxb.
In Vitro Immunomodulating Effect of C. zanthorrhiza
C. zanthorrhiza methanol extract has been reported to inhibit
ROS generation in a luminol and lucigenin-enhanced
chemiluminescence (CL) assay. C. zanthorrhiza rhizomes
reduced ROS production from whole blood of human by
in vitro study. Moreover, the extract significantly inhibited the
release of ROS from zymosan-induced PMNs and macrophages.
C. zanthorrhiza also showed strong inhibition on PMN
migration, with an IC
50
value of 2.5 μg/ml (Jantan et al.,
2011). A previous study reported that the methanol extract of
C. zanthorrhiza rhizomes showed strong inhibition on the
expression of CD18/11a; meanwhile, the extract has low effect
on leukocyte phagocytosis (Harun et al., 2015). The mRNA levels
of IL-1β, NF-κB p65, MMP-2, and MMP-8 on LPS-induced
human gingival fibroblast-1 cells were reduced after treatment
with crude polysaccharide extract of C. zanthorrhiza. The extract
of C. zanthorrhiza inhibited MAPK/activator protein-1 (AP-1)
signaling pathways. C. zanthorrhiza has been documented to
exhibit anti-inflammatory activities in LPS-induced RAW264.7
monocytes and H
2
O
2
-treated HT22 hippocampal cells (Kim
et al., 2018).
In Vivo Immunomodulating Effect of C. zanthorrhiza
Curcuminoid cider, a traditional fermented product made by the
addition of Acetobacter xylinum to curcuminoid fraction isolated
from C. zanthorrhiza, reduced the gene expression of IL1β,TNFα,
and chemokine in hypercholesterolemic rats (Hardiwati et al., 2019).
The data were in accordance with a previous study which
demonstrated the inhibitory activity of curcuminoid cider and
curcuminoid fraction from C. zanthorrhiza on the gene
expression of CD44, ICAM-1, iNOS, and LOX-1 in high-
cholesterol diet rats (Mauren et al., 2016). Volatile oil from C.
zanthorrhiza enhanced the lymphocyte proliferation from human
male B blood type (Miksusanti, 2012). C. zanthorrhiza extract
administration was able to reduce inflammatory lymphocytes in
alcohol-induced hepatitis in mice (Ilene et al., 2020). C. zanthorrhiza
ethanol extracts strongly reduced cytokine gene expression, which
include TNF-α,IL-6,IL-1β, and C-reactive protein (CRP) in the
liver, adipose tissue, and muscle of high-fat diet-induced obese mice
(Kim M-B et al., 2014). The crude polysaccharide extract of C.
zanthorrhiza consisted of glucose, galactose, arabinose, xylose,
mannose, and rhamnose, and was also reported to significantly
enhance the phagocytosis of macrophages and the production of
NO, H
2
O
2
,TNF-α,andPGE
2
. In addition, it clearly enhanced
phosphorylation of IκBα, suggesting a role as a NF-ĸBactivator(Kim
et al., 2007;Huang et al., 2010). C. zanthorrhiza–inhibited pro-
inflammatory cytokine production in mice induced high-fat diet. C.
zanthorrhiza extract at 100 mg/kg body weight/day decreased IL-1β
gene expression by 89.9% compared to the control group (Ilene et al.,
2020). C. zanthorrhiza was also reported to stimulate total and
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311911
Yuandani et al. Immunomodulatory Effects of Curcuma Species

differential leukocytes in African catfish (Clarias gariepinus)(Lestari
et al., 2019). C. zanthorrhiza rhizome in combination with Zingiber
officinale rhizome, Vitex trifolia leaves, Echinacea purpurea,and
citrus fruit in a herbal formula increased the number of macrophages
phagocytizing Candida albicans as compared to those of E.
purpurea–only group in mice. In addition, the herbal formula
also displayed immunostimulatory activities on lymphocyte
proliferation and the level of IgG actively phagocytizing C.
albicans (Ikawati et al., 2019).
Clinical Studies of C. zanthorrhiza on the Immune
System
An unsystematic clinical study of C. zanthorrhiza reported that C.
zanthorrhiza extract reduced the population of B lymphocytes
(Dewi et al., 2012). A previous study reported that C.
zanthorrhiza in combination with C. mangga and Phyllanthus
niruri maintained the levels of CD4
+
in HIV/AIDS patients
(Astana et al., 2018). C. zanthorrhiza in combination with
Vitex trifolia did not cause liver and kidney damage after
14 days, 3 times a day treatment in women (Baroroh et al.,
2011). Supplementation of C. zanthorrhiza with vitamin D3
was not able to decrease IL-6 level and elevate TGF-β1
systemic lupus erythematosus (SLE) in patients with
hypovitaminosis D (Wahono et al., 2017b). These data were
supported by a double-blind randomized controlled study on
active SLE patients with hypovitaminosis D, which reported that
addition of C. zanthorrhiza in vitamin D3 did not reduce IL-17
level as compared to those of singular vitamin D administration
(Wahono et al., 2017a). Furthermore, a placebo-controlled
double-blind clinical study showed that TNF-αrelease was
reduced after treatment with the extract of C. zanthorrhiza for
4 weeks in SLE patients (Setiawati et al., 2017).
C. zanthorrhiza Roxb. is the second most popular plant among
the genus Curcuma that has been investigated for its
immunomodulating properties. Similar to C. longa, the crude
extracts of C. zanthorrhiza were used in experimental studies to
evaluate its in vivo immunomodulating effect using various
animal models. There were a few in vitro studies, and the
chemical constituents of the extracts were mostly not
determined or the extracts were not standardized. Some
clinical trials have been conducted on C. zanthorrhiza extracts,
but they were unsystematic and not well designed. Despite the
regulatory requirements for clinical studies and sufficient data not
being generated on preclinical testing of C. zanthorrhiza, there
were already reports on a few unsystematic case studies to
evaluate the immunomodulating properties of C. zanthorrhiza
in human. For clinical studies, sufficient preclinical testing should
be generated using standardized extracts, which include
bioavailability, and pharmacokinetic and toxicological studies,
before they can be subjected to clinical studies.
Curcuma aeruginosa Roxb.
In Vitro Immunomodulating of C. aeruginosa
The methanol extract of C. aeruginosa at 100 and 6.25 μg/ml
showed moderate inhibition on CD18/11a expression on the
surface of phagocytes, which was determined using a flow
cytometry method. The extract at the same concentrations also
demonstrated low inhibition on phagocytosis of leukocytes
(Harun et al., 2015). Investigation on the effect of C.
aeruginosa methanol extract on ROS generation from
polymorphonuclear cells (PMNs) and peritoneal macrophages
in human whole blood revealed that the extract possessed ROS
inhibitory activity for luminol-stimulated chemiluminescence
(CL). C. aeruginosa rhizomes inhibited oxidative burst of
PMNs and macrophages, with IC
50
values of 1.8 and 4.6 μg/
ml, respectively. Interestingly, C. aeruginosa extract also
possessed significant ROS inhibitory activity for lucigenin-
enhanced CL. However, C. aeruginosa revealed low inhibition
on PMN chemotaxis toward the chemoattractant, N-formyl-
methionyl-leucyl-phenylalanine (fMLP), with percentage
inhibition of 49.9% (Jantan et al., 2011).
In Vivo Immunomodulating of C. aeruginosa
C. aeruginosa extract, obtained by steam distillation, has been
reported to increase the percentage of CD4
+
and CD8
+
cells
(Anggriani et al., 2019). A previous study reported that C.
aeruginosa ethanol extract was able to increase IFN-γ, TNF-α,
IL-2, and IL-12 levels in 7,12-dimethylbenz [a]anthracene
(DMBA)-induced Wistar rats. The highest stimulation on
cytokines release was shown after treatment with the ethanol
extract of C. aeruginosa at a dose of 80 mg/200 g body weight
(Sulfianti et al., 2019). The aqueous extract of C. aeruginosa in
combination with Piper retrofractum and Curcuma zanthorrhiza
supplemented in a fish fed at the concentrations of 0.5, 1, and
1.5%, respectively, enhanced nonspecific immunity of
Epinephelus fuscoguttatus. The addition of C. aeruginosa
extract induced significant difference in the total leukocyte
count of Epinephelus fuscoguttatus after being infected by
Vibrio alginolyticus and V. parahaemolyticus during 15 days of
observation. C. aeruginosa treatment increased the total leukocyte
count on day 4 and day 8. Moreover, C. aeruginosa at
concentration of 1% showed the strongest stimulation on
phagocytosis activity, which was determined on day 8 (Setyati
et al., 2019).
The in vitro and in vivo immunomodulating studies on C.
aeruginosa were carried out on their crude aqueous and ethanol
extracts. The bioactive metabolites contributing to the
modulating effects were not identified. It is important to
chemically characterize the extract to determine the bioactive
compounds contributing to the immunomodulatory properties
and mechanistic investigation to conclude the plant potency and
effects on the immune-related disorders.
Curcuma zedoaria (Christm.) Roscoe
In Vitro Immunomodulating Effect of C. zedoaria
C. zedoaria (Christm.) Roscoe rhizome extract has been
reported to inhibit NO production from LPS-stimulated
RAW264.7 cells. It has also been found to reduce iNOS and
COX-2 expressions (Lee et al., 2019). In another study, C.
zedoaria prevented ß-hexosaminidase release in RBL-2H3
cells and showed passive cutaneous anaphylaxis reaction in
mice. ß-Hexosaminidase is a marker of antigen-IgE–mediated
degranulation (Lobo et al., 2009). Essential oil from C. zedoaria
was reported to reduce TNF-αrelease from L. monocytogenes
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311912
Yuandani et al. Immunomodulatory Effects of Curcuma Species

and S. aureus–stimulated RAW264.7 cells (Huang et al., 2019).
Polysaccharide fraction of C. zedoaria rhizome was found to
enhance phagocytosis activity and splenocyte proliferation. It
also stimulated the primary and secondary titers as well as
delayed-type hypersensitivity response (Faradilla and Iwo,
2014). This work was supported by a previous study which
showed that polysaccharide fraction of C. zedoaria enhanced
phagocytosis of FITC-labeled Gram-negative bacteria (E. coli)
or Gram-positive bacteria (S. aureus) by peritoneal
macrophages. It also stimulated two microbicidal routes,
oxygen-dependent and oxygen-independent mechanisms.
Lysosomal activity increased after treatment with
polysaccharide fraction as well as in vivo and in vitro
respiratory burst. It was reported that PMA-induced
respiratory burst of peritoneal macrophage was higher than
those of RAW 264 cells identified using luminol-
chemiluminescence–based assay. The production of H
2
O
2
,
NO, and TNF-αwas also enhanced at the doses of 10, 50,
and 100 μg/ml, dose dependently (Kim et al., 2001).
In Vivo Immunomodulating Effect of C. zedoaria
The effect of C. zedoaria extract on tumor progression and
peripheral blood cells in C57Bl/6J mice injected with B16F10
murine melanoma cells was determined using different routes of
administration. A decrease in peritoneal cell number and a
significant increase in total red and white blood cell counts
were observed. Oral administration of the extract revealed a
noteworthy increase only in the total leukocyte count
(Carvalho et al., 2010). C. zedoaria has also been reported to
stimulate immune response in fish. Supplemented diets with C.
zedoaria increased the phagocytic rate and lysosome activity in
Epinephelus coioedes fish. C. zedoaria was able to increase reactive
oxygen production, identified using two different methods, NBT
test and chemiluminescent assay (Nan et al., 2014).
Similar to the other Curcuma species already discussed, the
metabolite profiles of C. zedoaria extracts were not determined. It
is necessary to analyze the chemical constituents of the extracts or
use standardized extracts in the studies as the phytochemical
constituents of the plant may vary with variation in genetic
adaptation of the plant population growing at different
altitudes, its geographical distribution due to the changes in
soil composition, and other environmental factors. Thus, using
standardized extracts will ensure the dynamic change of varying
amounts of phytochemical constituents in the plant is taken into
consideration.
Curcuma mangga Valeton & Zijp
In Vitro Immunomodulating Effect of C. mangga
A previous study reported in vitro NO inhibition activity of C.
mangga which might contribute to its anti-inflammatory effect
(Abas et al., 2006;Kaewkroek et al., 2009;Liu and Nair, 2011).
Furthermore, C. mangga rhizome extract and its chloroform,
hexane, and ethyl acetate fractions reduced NO production from
LPS-induced RAW 264.7 cells. Among the fractions, the
chloroform fraction showed the highest NO inhibition,
followed by hexane, and then ethyl acetate fractions
(Kaewkroek et al., 2009). A previous study of the methanol
extract of C. mangga rhizomes on whole blood showed that
the extract exhibited strong inhibitory activity upon activation
by zymosan. C. mangga rhizome extract possessed high ROS
inhibitory activity in PMNs and peritoneal macrophages as
investigated in a luminol-enhanced CL assay. The extract also
inhibited the release of ROS from PMNs and macrophages in a
lucigenin-enhanced CL assay, with IC
50
values of 0.9 and 6.6 μg/
ml, respectively (Jantan et al., 2011). C. mangga methanol extract
has also been found to significantly suppress the cell surface
expression of CD18/11a as compared to the negative control.
However, the extract of C. mangga rhizome at the concentration
of 100 and 6.25 μg/ml showed immunostimulatory activity on
phagocytosis of leukocytes (Harun et al., 2015).
In Vivo Immunomodulating Effect of C. mangga
C. mangga Valeton & Zijp rhizome ethanol extract, its different
organic fractions (hexane, chloroform, and ethyl acetate), and
aqueous fraction have showed appreciable anti-inflammatory and
analgesic activities in mice and inflammatory models using croton oil-
inducedmouseearedemaandcarrageenan-induced rat paw edema.
The plant extract and its fractions at 200 mg/kg demonstrated
analgesic activity by reducing the number of writhing and also
produced antinociception using hot plate and formalin test. At
200 mg/kg, the hexane and chloroform fractions significantly
prolonged the latency time, but ethyl acetate and aqueous fractions
were not active. In addition, the ethanol extract of C. mangga rhizome
and its fractions displayed significant reduction of paw and ear edema
in rat (Ruangsang et al., 2010). Our previous study reported that the
n-hexane, ethyl acetate, and ethanol extracts of C. mangga rhizomes at
the doses of 100, 200, and 400 mg/kg increased the carbon clearance
rate, indicating the enhancement of carbon engulfment by cells in the
reticuloendothelial system of mice, thus stimulating the phagocytosis
activity in mice (Yuandani and Suwarso, 2017a;Yuandani et al., 2019).
In addition, the C. mangga rhizome ethanol extract exhibited
stimulation of antibody titer against bovine red blood cells in a
dose-dependent way by using the hemagglutination method. The
cellular immunity was also enhanced after treatment with C. mangga
ethanol extract by increasing the bovine red blood cell–induced mice
paw volume (Yuandani et al., 2018). Moreover, the ethanol extract of
C. mangga rhizome stimulated the immune response in doxorubicin-
induced immunosuppressive rats, which was indicated by the
elevation of antibody titer and delayed hypersensitivity (DTH)
response (Yuandani et al., 2020).
As with other Curcuma species already discussed, the effects of
C. mangga on the immune cells and experimental animals may
vary considerably, depending on the experimental conditions
used, including the solvent of extraction, extraction method, cell
line, animal model, treatment scheme, and different disease
animal models. Dosage and concentration of raw extracts of
the plant are crucial in order to achieve the desired benefit. Thus,
to ensure the results are reproducible when the study is replicated,
the same methodology has to be used by other researchers.
Curcuma amada Roxb.
The ethanol, petroleum ether, chloroform, and acetone extracts of
C. amada enhanced the phagocytosis activity of PMNs. The
ethanol extract at a concentration 3 mg/ml showed the highest
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311913
Yuandani et al. Immunomodulatory Effects of Curcuma Species

TABLE 2 | Bioactive compounds of Curcuma species with immunomodulating activity and their mechanisms of action.
Main compound Species Subjects Study
design
Immunomodulatory
activities
Modulation Parameters/
mediators
affected
References
Curcumin Curcuma
species
High glucose-
cultured monocytes
in vitro Cytokine production ↓IL6, IL8, TNFα,
and MCP1
Jain et al. (2009)
Streptozotocin-
induced rats
in vivo Cytokine production ↓IL6, TNFα,and
MCP1
Jain et al. (2009)
Mice pancreatic in vivo Leukocyte infiltration ↓Leukocytes Castro et al. (2014)
M-stimulated
BDC2.5-splenocytes
in vitro T-cell proliferation ↓CD4
+
, T cells,
and IFN-γ
Castro et al. (2014)
BDC2.5 mice T
lymphocite
in vitro T-cell proliferation ↓T lymphocyte Castro et al. (2014)
PMN leukocytes in vitro DHA synthesis ↑DHA Pisani et al. (2009),
Wu et al. (2015)
PMN leukocytes in vitro ROS production ↓ROS Pisani et al. (2009),
Wu et al. (2015)
LPS-induced mice
mastitis
in vivo Myeloperoxidase
activity
↓MPO Fu et al. (2014)
LPS-induced mice
mastitis
in vivo Cytokine production ↓TNF-α, IL-6, IL-
1β, and TLR4
Fu et al. (2014)
LPS-induced mice
mastitis
in vivo Phosphorylation ↓IκB-αand NF-
κB p65
Fu et al. (2014)
Microglial cells in vitro NO production ↓NO Cianciulli et al.
(2016)
Microglial cells in vitro Phosphorylation ↓IL-1β, IL-6, TNF-
α, and PI3K/Akt
Cianciulli et al.
(2016)
Microglial cells in vitro NF-κB and iNOS
expression
↓NF-κB and iNOS Cianciulli et al.
(2016)
Microglial cells in vitro Cytokine production ↓NO, PGE
2
, TNF-
α, iNOS, and
COX-2
Yu et al. (2018)
C. longa Healthy albino mice in vivo White blood cells
production and weight
lymphoid
↑Lymphoid organs
and white blood
cells
Afolayan et al.
(2018)
Dendritic cells in vitro Surface molecule
expression
↓CD80, CD86,
MHC class II, and
IL-1
Kim et al. (2005)
Dendritic cells in vitro Cytokine production ↓IL-6, IL-12, and
TNF- α
Kim et al. (2005)
Dendritic cells in vitro NF-κB p65
translocation
↓NF-κB p65 Kim et al., 2005
Bronchoalveolar of
Balb/c mice
in vivo Allergic response ↓Eosinophils Ravikumar and
Kavitha (2020)
Bronchoalveolar of
Balb/c mice
in vivo Cytokine production ↓IL-4 Ravikumar and
Kavitha (2020)
PBMCs in vitro T-cell proliferation ↓Lymphocyte Yadav et al. (2005)
PBMCs in vitro Cytokine production ↓IL-2 and TNF-αYadav et al. (2005)
PBMCs in vitro NF-κB↓NF-κBYadav et al. (2005)
Erythroleukemic cell
line K562
in vitro Cytotoxicity ↑NK cell Yadav et al. (2005)
Lupus BALB/c mice in vivo Adaptive immune
response
↓Th1, Th2, and
Th17
Kalim et al. (2017)
Lupus BALB/c mice in vivo ANA levels ↓ANA Kalim et al. (2017)
Monocytes and liver
macrophages
in vivo ROS production ↓ROS Inzaugarat et al.
(2017)
Monocytes in vivo TNF-αand IFN- γ
production
↑TNF-αand IFN- γInzaugarat et al.
(2017)
C. longa Fish in vivo Immune response ↑Immune Alambra et al.
(2012)
C. zedoaria RBL-2H3 cells in vitro beta-Hexosaminidase
production
↓Beta-
hexosaminidase
Matsuda et al.
(2004)
RBL-2H3 cells in vitro Cytokine production ↑TNF–αand IL–4Matsuda et al.
(2004)
Turmeronol C. longa RAW264.7 cells in vitro PGE
2
and NO
production
↓PGE
2
and NO Okuda-Hanafusa
et al. (2019)
(Continued on following page)
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311914
Yuandani et al. Immunomodulatory Effects of Curcuma Species

TABLE 2 | (Continued) Bioactive compounds of Curcuma species with immunomodulating activity and their mechanisms of action.
Main compound Species Subjects Study
design
Immunomodulatory
activities
Modulation Parameters/
mediators
affected
References
RAW264.7 cells in vitro Cytokine production ↓IL-1βand IL-6 Okuda-Hanafusa
et al. (2019)
Cytoplasm into the
nucleus
in vitro NF-κB translocation ↓NF-κBOkuda-Hanafusa
et al. (2019)
Curdione C.
aeruginosa
CD95 protein in silico Docking score ↓Curdione to
CD95
Anggriani et al.
(2019)
1,8-cineol CD95 protein in silico Docking score ↑1,8-cineol to
CD95
Anggriani et al.
(2019)
Isocurcumenol Chicken embryo
fibroblast
in vitro Toxicity - Fibroblast cells
and lymphocytes
Lakshmi et al.
(2011)
Isoprocurcumenol RAW264.7 cells in vitro NO activity ↓NO Lee et al. (2019)
Germacrone C. zedoaria RAW264.7 cells in vitro NO activity ↓NO Lee et al. (2019)
Curzerenone RAW264.7 cells in vitro NO activity ↓NO Lee et al., 2019
Curcumenol RAW264.7 cells in vitro NO activity ↓NO Lee et al. (2019)
Curcuzedoalide RAW264.7 cells in vitro NO activity ↓NO Lee et al. (2019)
RAW264.7 cells in vitro iNOS and COX-2
response
↓iNOS and COX-2 Lee et al. (2019)
Dihydrocurcumin RBL-2H3 cells in vitro beta-Hexosaminidase
production
↓beta-
Hexosaminidase
RBL-2H3 cells in vitro Cytokine production ↑TNF-αand IL-4 Matsuda et al.
(2004)
Tetrahydrodemethoxycurcumin RBL-2H3 cells in vitro beta-Hexosaminidase
production
↓beta-
Hexosaminidase
Matsuda et al.
(2004)
RBL-2H3 cells in vitro Cytokine production ↑TNF-αand IL-4 Matsuda et al.
(2004)
Tetrahydrobisdemethoxycurcumin RBL-2H3 cells in vitro Hexosaminidase
production
↓beta-
Hexosaminidase
Matsuda et al.
(2004)
RBL-2H3 cells in vitro Cytokine production ↑TNF-αand IL-4 Matsuda et al.
(2004)
1,7-bis(4-hydroxyphenyl)-1,4,6-
heptatrien-3-one
Lipopolysaccharide
(LPS)-activated
macrophages
in vitro TNF-αproduction ↓TNF-αJang et al. (2001)
Macrophages in vitro NO production and
iNOS expression
↓NO and iNOS Jang et al. (2004)
Procurcumenol lipopolysaccharide
(LPS)-activated
macrophages
in vitro TNF-αproduction ↓TNF-αJang et al. (2001)
Xanthorrhizol C.
zanthorrhiza
Human gingival
fibroblast-1 cells
in vitro mRNA levels ↓IL-1β,NF-κB
p65, MMP-2,
and MMP-8
Kim et al. (2018)
RAW 264.7 cell line in vitro MAPK and AP-1
response
↓MAPK and AP-1 Kim et al. (2018)
Demethoxycurcumin C. mangga RAW 264.7 cell line in vitro NO production ↓NO Kaewkroek et al.
(2009)
RAW 264.7 cell line in vitro NO and PGE
2
production
↓NO and PGE
2
Kaewkroek et al.
(2010)
RAW 264.7 cell line in vitro mRNA expressions ↓iNOS and COX-2 Kaewkroek et al.
(2010)
Bisdemethoxycurcumin RAW 264.7 cell line in vitro NO production ↓NO Kaewkroek et al.
(2009)
RAW 264.7 cell line in vitro NO and PGE
2
production
↓NO and PGE
2
Kaewkroek et al.
(2010)
RAW 264.7 cell line in vitro mRNA expressions ↓iNOS and COX-2 Kaewkroek et al.
(2010)
4-[(1R, 4aR, 8aR)-decahydro-5, 5,
8a-trimethyl-2-methylene-1-
naphthalenyl]-, (3E)-rel
RAW 264.7 cell line in vitro NO and PGE
2
production
↓NO and PGE
2
Kaewkroek et al.
(2009)
RAW 264.7 cell line in vitro mRNA expressions ↓iNOS and COX-2 Kaewkroek et al.
(2009)
15,16 bisnorlabda-8(17), 11-dien-
13-one
RAW 264.7 cell line in vitro NO and PGE
2
production
↓NO and PGE
2
Kaewkroek et al.
(2010)
(Continued on following page)
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311915
Yuandani et al. Immunomodulatory Effects of Curcuma Species

stimulation on percentage of phagocytosis. Further study on
delayed hypersensitivity response against sRBCs showed that
the ethanol extract of C. amada increased the paw volume.
Moreover, the ethanol extract at the doses of 100, 200, and
400 mg/kg enhanced the antibody titer dose-dependently
(Karchuli and Pradhan, 2011). Supercritical carbon dioxide
(CO
2
) extract prepared from C. amada rhizomes has potential
to be used for the treatment of immune disorder such as
autoimmune diseases. Specifically, the extract can be used to
treat or prevent hypersensitivity diseases, in particular IgE-
mediated allergic reactions as well as autoimmune disorders
(Weidner et al., 2001). C. amada in combination with
Tinospora cordifolia,Piper longum, and Albizia lebbeck in a
herbal preparation can be used to treat allergy (Palpu et al.,
2008). The chemical constituents responsible for eliciting the
activity were not determined, although a few potent activities
have been reported on C. amada extract. There is a need to
proceed to study in detail the underlying mechanisms on relevant
signaling events followed by in vivo studies to explore the
potential of this plant as a natural immunomodulating agent.
Immunomodulatory Effects of Bioactive
Compounds of Curcuma Species
Plants in the genus Curcuma contain many compounds which
contribute to the immunomodulatory activity of the plants, as
shown in Table 2. Among the compounds from Curcuma
species, curcumin and xanthorrhizol have been discussed in detail
in this review as they have been widely investigated for their
immunomodulating effects on the innate and adaptive immune
system. Other compounds including turmeronols, curdione,
curcuzedoalide, demethoxycurcumin, bisdemethoxycurcumin,
dihydrocurcumin, curcumenol, epi-procurcumenol,
isocurcumenol, and iso-procurcumenol germacrone are also
included in this review, but their data are limited as they have
not been well investigated for their immunomodulating effects. The
chemical structures of these compounds are included in Figure 2.
Curcumin
It is a major compound of C. longa and can also be found in other
Curcuma species. This natural diarylheptanoid compound has
been mainly isolated from the rhizomes of C. longa and studied
extensively for various pharmacological activities, including
antioxidant, anti-inflammatory, immunomodulatory,
antiangiogenic, anticancer, antiproliferative, and proapoptotic.
It has been one of the most intensively investigated compounds
for its immunomodulatory properties. Many preclinical
investigations which include in vitro cell assays and in vivo
studies in animal models have been carried out on curcumin
to evaluate its modulatory effects in the immune system. It is also
undergoing extensive clinical trials based on its anti-
inflammatory properties for the treatment of cancer.
In Vitro Immunomodulating Effect of Curcumin
The immunomodulating activity of curcumin has been
demonstrated by many in vitro studies using several immune
cells. Curcumin has been shown to inhibit inflammatory
responses by suppressing COX-2 and NO, NF-ĸB, iNOS, and
lipoxygenase in IFN-γor TNF-α–activated macrophages and NK
cells (Surh et al., 2001). A study by Jain et al. (2009) revealed that
curcumin significantly reduced the production of IL-6, IL-8,
TNF-α, and MCP-1 from high glucose-cultured monocytes.
Low concentration of curcumin reduced NOS activity and NO
production from macrophages. In another study, curcumin
inhibited the immunostimulatory function of dendritic cells,
leading to the reduction of CD80, CD86, and MHC class II
expression, but not MHC class I expression as well as IL-12
expression and cytokine release (IL-1, IL-6, and TNF- α).
Curcumin also inhibited LPS-induced MAPK activation and
the translocation of NF-B p65 as well as impaired induction of
Th1 responses (Kim et al., 2005). Gao et al. (2004) demonstrated
that curcumin inhibited pro-inflammatory cytokine (TNF-α, IL-
1, IL-12, and IL-6) expression in PMA or LPS-activated
macrophages, dendritic cells, monocytes, and splenic
lymphocytes. Curcumin has also been found to suppress PHA-
stimulated lymphocyte proliferation and IL-2 release as well as
transcription factor NF-KB and TNF-αproduction from LPS-
stimulated PBMC to enhance NK cell cytotoxicity (Yadav et al.,
2005). Curcumin has been shown to be able to increase ω-3
polyunsaturated fatty acid (PUFA) synthesis in the brain. An
in vitro study has showed that PUFAs have beneficial effects on
stimulating immune response by stimulating neutrophil
phagocytosis activity, while decreasing the release of ROS in
goat neutrophils (Pisani et al., 2009;Wu et al., 2015).
TABLE 2 | (Continued) Bioactive compounds of Curcuma species with immunomodulating activity and their mechanisms of action.
Main compound Species Subjects Study
design
Immunomodulatory
activities
Modulation Parameters/
mediators
affected
References
RAW 264.7 cell line in vitro mRNA expressions ↓iNOS and COX-2 Kaewkroek et al.
(2010)
(E)-15,15-diethoxylabda-8 (17),12-
dien-16-al
RAW 264.7 cell line in vitro NO and PGE
2
production
↓NO and PGE
2
Kaewkroek et al.
(2010)
RAW 264.7 cell line in vitro mRNA expressions ↓iNOS and COX-2 Kaewkroek et al.
(2010)
↑, increase.
↓, decrease.
-, no changes.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311916
Yuandani et al. Immunomodulatory Effects of Curcuma Species

Curcumin also increased ROS production from linoleic
acid–stimulated monocytes and liver macrophages as well as
TNF-αproduction from leptin-induced monocytes and IFN-γ
release from CD4
+
T cells (Inzaugarat et al., 2017). Pretreatment
with curcumin significantly reduced the production of NO, the
expression and release of TNF-α, IL-1β, IL-6, PI3K/Akt
phosphorylation, NF-κB activation, and iNOS expression in
LPS-stimulated microglial cells (Cianciulli et al., 2016). In
addition, NO, PGE
2
, and TNF-αproduction as well as iNOS
and COX-2 expression in LTA-activated microglial cells were
reduced by curcumin (Yu et al., 2018). In an in silico study,
curcumin has been shown to bind to viral S1 protein, which is
important for SARS-CoV-2 entry; hence, it may prevent cytokine
storm in a severe form of COVID-19 (Pawitan, 2020). The
pathways and inflammatory mediators involved in the
immunomodulating property of curcumin are illustrated in
Figure 3.
In Vivo Immunomodulating Effect of Curcumin
In vivo studies to determine the immunomodulatory effects of
curcumin were carried out using several animal models.
Curcumin reduced the levels of IL6, TNFα, and MCP1 in
streptozotocin-induced type 1 diabetes rats. Moreover,
curcumin prevented pancreatic leukocytes infiltration that
might initiate ß-cell destruction. In addition, curcumin
reduced CD4
+
T cell proliferation and IFN-γrelease from
M-stimulated BDC2·5-splenocytes as well as reduced LPS/IFN-
γ–induced dendritic cell maturation. Antigen-specific
T-lymphocyte proliferation has also been reduced by curcumin
action on both T cells and antigen-presenting cells (APCs)
(Castro et al., 2014). Administration of curcumin to lactating
mice prevented mice mastitis by reducing the MPO activity;
expression of TNF-α, IL-6, IL-1β, TLR4; and phosphorylation
of IκB-αand NF-κB p65 after being induced by LPS (Fu et al.,
2014). Nanoparticulate curcumin demonstrated stronger activity
on cellular and humoral immunity, and increased lymphoid
organs and white blood cell production than those of control
(Afolayan et al., 2018). A previous study reported that curcumin
displayed immunomodulatory effects in comorbid diabetic
asthma mice by reducing eosinophil number and Il-4 level
with a high IFN-γto IL-4 ratio in the blood and
bronchoalveolar after ovalbumin injection (Ravikumar and
Kavitha, 2020). Administration of curcumin to pristane-
induced lupus mice decreased Th1, Th2, and Th17 and
slightly increased Treg percentages. In addition, ANA levels
were also decreased after curcumin treatment (Kalim et al.,
2017). Curcumin was able to enhance immune response in
Macrobrachium rosenbergii after being challenged with Vibrio
alginolyticus (Alambra et al., 2012).
Clinical Studies of Curcumin on Immune System
Presently, curcumin is under extensive clinical investigation
where there are 116 ongoing clinical trials on curcumin, the
status of which can be found on http://www.clinicaltrials.gov/.
Among the clinical trials on curcumin, 99 of them were based on
its anti-inflammatory properties. Cancer (e.g., breast, pancreatic,
FIGURE 2 | Chemical structures of potential immunomodulators from Curcuma species.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311917
Yuandani et al. Immunomodulatory Effects of Curcuma Species

lung, colorectal, and prostate), inflammatory bowel diseases
(IBD; Crohn’s disease and ulcerative colitis) and rheumatoid
arthritis were the major diseases for which trials had been
conducted, reflecting the pleiotropic actions of curcumin. In
these trials, curcumin often acted as a dietary supplement or
an adjunct treatment to the standard therapy. Studies on the
efficacy and safety of curcumin as adjuvant in the treatment of
cancer and cognitive damage will continue to dominate in future
clinical trials (Jurenka, 2009). In their review on the clinical effects
of curcumin in ulcerative colitis, Kumar et al. (2012) suggested
that curcumin may be a safe and effective therapy for the
maintenance of remission when given as adjunct therapy in
quiescent ulcerative colitis. However, the results were
preliminary due to the low number of enrolled patients
participated in the clinical trials, as suggested by Fürst and
Zündorf (2014). They suggested more thorough controlled
randomized trials are required to be pursued to determine the
safety level and efficacy of the compound for human use. The
success of curcumin as a potent anti-inflammatory agent in future
depends on the findings of high-quality and big cohort studies.
Moreover, curcumin has poor bioavailability, and many studies
have been carried out to address this issue via chemical and
technological methods. Preparation of more stable curcumin
derivatives and use of nanotechnology for curcumin delivery
are actively being pursued to improve the bioavailability of
curcumin (Anand et al., 2007). Curcumin still has potential to
be used clinically for the treatment of the abovementioned
indications as it is nontoxic with good safety profile and well
tolerated.
Xanthorrhizol
Xanthorrhizol is a bisabolane-type sesquiterpenoid, isolated from
C. zanthorrhiza Roxb. It is known to possess diverse
pharmacological activities, including antioxidant, anti-
inflammatory, antimicrobial, anticancer, hepatoprotective,
nephroprotective, antihypertensive, antihyperglycemic,
antiestrogenic, and antiplatelet effects. Xanthorrhizol reduced
mRNA levels of MMP-2, MMP-8, NF-κB p65, and IL-1βin
LPS-induced human gingival fibroblast-1 cells. The compound
also has anti-inflammatory activities in H
2
O
2
-treated HT22
hippocampal cells and inhibited MAPK/activator protein-1
(AP-1) signaling pathways (Kim et al., 2018). In a study to
evaluate the effects of standardized C. zanthorrhiza extract and
its marker compound, xanthorrhizol, on hyperglycemia and
FIGURE 3 | Modulatory effects of curcumin on the NF-κB, MAPK, and Akt signaling pathways. The thick red block sign indicates the possible point of modulation of
the signal transduction pathways. NF-κB, nuclear factor kappa β; MAPK, mitogen-activated protein kinase; PI3K/Akt, phosphatidylinositol 3-kinase and protein kinase B;
P, phosphoryl group.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311918
Yuandani et al. Immunomodulatory Effects of Curcuma Species

inflammatory markers in high-fat diet–induced obese mice,
xanthorrhizol was found to significantly inhibit inflammatory
cytokine release, such as IL-1β, IL-6, TNF-α, and C-reactive
protein (CRP), in the liver, muscle, and adipose tissue (Kim
M-B et al., 2014).
Other Compounds
In addition to curcuminoids, sesquiterpenes like turmeronols
have potential to suppress the immune response. Turmeronol A
and turmeronol B from C. longa were tested on mouse
macrophages (RAW 264,7 cells). Both turmeronols showed
inhibitory of PGE
2
and NO production as well as IL-6 and IL-
1βat the mRNA and protein levels in LPS-induced cells.
Turmeronols also inhibited the translocation of NF-κB from
the cytoplasm into the nucleus (Okuda-Hanafusa et al., 2019).
Curdione, a major compound of C. aeruginosa, has been
proposed as a potential immunomodulatory agent. Molecular
docking analysis revealed that there is a high probability of
interaction between curdione and CD95 protein, as the
replacement of native ligand. The docking score of curdione to
the protein was lower than that of the native ligand. In addition,
1,8-cineol from C. aeruginosa also has a high docking score to
CD95 protein, but not significant as compared to those of native
ligand of CD95 (Anggriani et al., 2019). Isocurcumenol isolated
from C. zedoaria did not demonstrate any significant toxicity on
normal chicken embryo lymphocytes and fibroblast cells
(Lakshmi et al., 2011). A recent study showed that five
sesquiterpenoids (isoprocurcumenol, germacrone, curzerenone,
curcumenol, and curcuzedoalide) from C. zedoaria demonstrated
inhibitory activity on NO synthesis. Among the compounds,
curcuzedoalide showed the highest inhibition. Further study
showed that curcuzedoalide inhibited the expression of pro-
inflammatory mediators (iNOS and COX-2) (Lee et al., 2019).
Curcumin, dihydrocurcumin, tetrahydrodemethoxycurcumin,
and tetrahydrobisdemethoxycurcumin in C. zedoaria enhanced
the release of IL-4 and TNF–α, and inhibited the production of ß-
hexosaminidase. ß-Hexosaminidase is a marker of antigen-
IgE–mediated degranulation (Putri, 2014).
Isolated compounds from C. zedoaria, that is,
epiprocurcumenol, procurcumenol, and 1,7-bis(4-
hydroxyphenyl)-1,4,6-heptatrien-3-one, inhibited the
production of TNF-αfrom LPS-activated macrophages (Jang,
et al., 2001). These compounds, especially 1,7-bis(4-
hydroxyphenyl)-1,4,6-heptatrien-3-one, were also found to
exhibit strong inhibition against production of NO and
expression of iNOS in activated macrophages (Jang et al.,
2004). Demethoxycurcumin, bisdemethoxycurcumin, and 3-
buten-2-one, 4-[(1R, 4aR, 8aR)-decahydro-5, 5, 8a-trimethyl-2-
methylene-1-naphthalenyl]- (3E)-rel isolated from C. manga
reduced the production of NO from LPS-stimulated RAW
264.7 cells. Among the compounds tested,
demethoxycurcumin showed the highest NO inhibition
(Kaewkroek et al., 2009). In an effort to elaborate the anti-
inflammatory mechanism of compounds from C. mangga
rhizomes, Kaewkroek et al. (2010) evaluated the anti-
inflammatory effects of several compounds against production
of PGE
2
and NO from RAW 264.7 cells. These include
demethoxycurcumin, bisdemethoxycurcumin,
15,16 bisnorlabda-8 (17),11-dien-13-one, and (E)-15,15-
diethoxylabda-8 (17),12-dien-16-al. Of all the compounds
tested, 11-dien-13-one (E)-15,15-diethoxylabda-8 (17),12-dien-
16-al demonstrated the strongest NO inhibitory activity, while
demethoxycurcumin displayed the strongest activity on PGE
2
release. Moreover, investigation of mechanism at the
transcriptional level showed that all the compounds reduced
the mRNA expressions of COX-2 and iNOS, except
15,16 bisnorlabda-8 (17), 11-dien-13-one, which only
downregulated the mRNA of iNOS.
Most of the studies on the bioactive secondary metabolites of
Curcuma species were carried out at cellular and molecular levels
on various immune cells to explore their effects on the release and
expression of pro-inflammatory mediators via various signaling
pathways, such as NF-κB, MAPKs, and other events. More in-
depth studies to understand the underlying mechanisms using
experimental in vivo animal models of immune-related disorders
and elaborate bioavailability, preclinical pharmacokinetics, and
toxicity studies are required before clinical trials can be pursued
for development into immunomodulatory agents.
TOXICOLOGICAL STUDIES
Systematic safety evaluations and toxicological investigations on
C. longa and curcumin have indicated that they are nontoxic for
human consumption, especially by oral administration. It is
considered non-genotoxic, non-mutagenic, and generally
recognized as safe. Several studies have indicated that oral
administration of C. longa and curcumin in animals was safe
without reproductive toxicity at certain doses. Clinical trials have
indicated that the safe dose for human consumption was at an
oral dose of 6 g/day for 4–7 weeks. In rare cases, minor side effects
like gastrointestinal upsets may happen (Soleimani et al., 2018). A
cheminformatics approach was used to predict toxicity, which
includes human hepatotoxicity, rodent carcinogenicity, and
bacterial mutagenicity of 200 chemical compounds found in C.
longa. Of the compounds studied, 136 compounds were predicted
as mutagenic, 184 were toxigenic, 64 were hepatotoxic, and 153
were carcinogenic. Interestingly, a dose-dependent hepatotoxicity
may occur with curcumin and its derivatives. The study also
predicted that few other constituents of C. longa are
noncarcinogenic, non-mutagenic, non-hepatotoxic, and devoid
of any side effects (Balaji and Chemakam, 2010).
Liju et al. (2013) reported the acute and sub-chronic toxicity
studies of the essential oil of C. longa (EOCL). For the acute
toxicity test, up to 5 g of EOCL per kg body weight was
administered in a single dose to Wistar rats, while for the
subacute toxicity study, the rats were administered with a
daily oral administration at doses of 0.1, 0.25, and 0.5 g/kg for
13 weeks. The results indicated that the EOCL was nontoxic as
there were no changes in body weight, and no mortality or
adverse clinical signs during both acute and sub-chronic
toxicity studies. The hepatic function was normal, and the
biomarkers, alanine amino transferase (ALT), alkaline
phosphatase (ALP), and aspartate aminotransferase (AST)
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311919
Yuandani et al. Immunomodulatory Effects of Curcuma Species

remained unchanged in treated animals. There was no subacute
toxicity as triglycerides, total cholesterol, serum electrolyte
parameters, histopathology of tissues, and markers of renal
function remained unchanged after 13 weeks of treatment
with curcumin. There was no mutagenicity to Salmonella
typhimurium up to 3 mg/plate. Oral administration of 1 g/kg
body weight EOCL for 14 days did not produce any genotoxicity
as there was no DNA damage and chromosome aberration or
micronuclei in rat bone marrow cells (Mary et al., 2012).
The acute toxicity study of C. zanthorrhiza ethanol extract at
5000 mg/kg revealed that the extract did not show any toxicity
signs such as salivation, sleeping, diarrhea, or lethargy in mice
(Devaraj et al., 2010). The result was in accordance with a
previous study which showed no toxicity sign was observed in
rats after administration of C. zanthorrhiza ethanol extract at
2000 and 5000 mg/kg. During 14 days of observation, rats showed
no clinical toxic signs, such as hypoactivity, hyperactivity,
lethargy, dermatitis, anorexia, depression, and jaundice as well
as no abnormalities in the kidney and liver (Rahim et al., 2014).
Listyawati (2006) reported the chronic toxicity study of ethanol
extract of C. zanthorrhiza. The extract at 50 mg/kg/day did not
induce significant effects on spermatogenic and hematological
changes (Listyawati, 2006). C. zanthorrhiza supplement at
2000 mg/kg showed no significant abnormalities on the lung,
heart, liver, kidney, and stomach. The LD
50
of C. zanthorrhiza
supplement as hepatoprotective was greater than 5000 mg/kg bw
(Arifin et al., 2020). Based on a clinical study on 30 healthy
subjects, the administration of C. zanthorrhiza in combination
with Vitex trifolia at doses of 1,500 and 4,500 mg/day for 14 days
did not alter the liver and kidney function, while at a dose of
9000 mg/day, the administration altered the AST and serum
creatinine values, indicating the extract affected the liver and
kidney functions (Baroroh et al., 2011). An aqueous extract of C.
zanthorrhiza at 2000 and 5000 mg/kg body weights also showed
no toxicity in mice or rats. Xanthorhizol, the active constituent of
C. zanthorrhiza, at a dose of 500 mg/kg did not cause mortality in
mice (HMPC, 2014).
Our previous study reported the acute toxicity evaluation of
ethanol extract of C. mangga rhizomes. Mice were administered with
the extract at 500, 1000, 2000, and 5000 mg/kg body weights as a
single dose, followed by 14 days of observation. Signs of toxicity were
revealed as lethargy was observed after treatment with C. mangga
extract at doses of 2000 and 5000 mg/kg body weight. Meanwhile,
other signs of toxicity such as diarrhea, coma, and salivation were not
recorded. In addition, the extract did not cause deleterious effect on
mice body weight. Macroscopic examination of two main organs
(liver and kidney) showed that the texture and color of both organs
were comparable to those of normal group. C. mangga extract at the
dose of 5000 mg/kg caused sinusoidal dilation in the liver and
glomerular lesion in the right kidney; however, there was no
lesion in the left kidney. The extract at the highest dose did not
cause mortality; hence, it can be considered that the LD
50
of C.
mangga extract was estimated to be more than 5000 mg/kg body
weight (Yuandani and Suwarso, 2017a).
The cytotoxicity evaluation of C. aeruginosa rhizomes on
fibroblast test has been conducted by Yuliawati and Hestianah
(2010). The results revealed that the extract at the concentrations
ranging from 1 to 25 ppm was not toxic, as indicated by the
percentage of cell viability ranging from 81.60 to 90.57%
(Yuliawati and Hestinah, 2010). Sub-chronic toxicity evaluation
of C. aeruginosa starch was conducted in Wistar rats. C.
aeruginosa starch was administered daily for 90 days. The
observation was performed for 90 days and followed until
120 days for satellite group to evaluate the reversible or
irreversible effect. The hematological parameters were observed,
these include leukocytes, hemoglobin, red blood cell (RBC),
hematocrit, mean corpuscular hemoglobin concentration
(MCHC), mean corpuscular hemoglobin (MCH), mean
corpuscular volume (MCV), and platelet levels. The results
indicated that there was no toxic effect on hematological
parameters (Kusumarin et al., 2020). C. aeruginosa in
combination with Allium sativum,Terminalia bellirica,and
Amomum compactum has been evaluated for their safety. Acute
toxicity evaluation has been performed according to the fixed-dose
method of OECD guideline 420. The herbal formulation did not
cause any toxic signs and symptoms. The were no abnormalities
found on body weight gain, macroscopic and microscopic
examinations as well as the relative organ weight after treatment
with the herbal preparation at the doses of 300 and 2000 mg/kg body
weight. Further study on sub-chronic toxicity showed that the
extracts did not induce any physical toxic symptoms as well as
abnormal weight gain and hematological parameters. Moreover, the
herbal formulation at a dose of 4,032 mg/kg did not cause any toxic
effects on the liver and kidney, which was indicated by the normal
values of urea, creatinine, total protein, albumin, globulin, aspartate
aminotransferase (AST) or glutamic oxaloacetic transaminase
(GOT), and alanine aminotransferase (ALT). In addition,
macroscopic and microscopic examinations showed that there
were no toxic effect on all organs tested (Sholikhah et al., 2020).
The evaluation of acute toxicity study of the purified fraction
of C. zedoaria revealed that the fraction at the dose of 41.6 and
35.7 mg/kg did not significantly alter the liver and kidney enzyme
levels. The LD
50
was 500 mg/kg bw (Lakshmi et al., 2011).
Furthermore, C. zedoaria ethanol extract at 150 mg/kg/day
revealed a significant reduction of RBC, Hb level, and
spermatozoa quality in chronic toxicity evaluation (Listyawati,
2006). The essential oil of C. zedoaria at 100 or 200 mg/kg
revealed weight loss, and abnormal hematological and
biochemical changes on dams and embryos in GD17 pregnant
rats. The toxicity mechanism may be related to placental
calcification in angiogenesis (Zhou et al., 2013). Sudeepthi
et al. (2014) reported the safety evaluation of C. amada
rhizomes in short-term treatment. The acetone extract of C.
amada (500–2000 mg/kg) was administered to the test
animals. The results indicated that the highest dose tested did
not cause mortality (Sudeepthi et al., 2014).
CONCLUSION AND FUTURE DIRECTIONS
In the last 20 years, many plants of the genus Curcuma especially
C. longa,C. zanthorrhiza,C. amada,C. mangga,C. aeruginosa,
and C. zedoaria and some of their bioactive compounds have
been investigated for their immunomodulating effects on the
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311920
Yuandani et al. Immunomodulatory Effects of Curcuma Species

immune system. Most of the studies were in vitro and in vivo and
only a few of the preclinical studies have progressed into clinical
studies. The up-to-date literature gathered indicated that the
immunological investigations on the plant extracts were
mainly preliminary with little mechanistic studies. Most of the
studies were on the crude extracts of the rhizomes. The extracts
were not appropriately characterized chemically or standardized
to the bioactive marker compounds which were responsible for
the activity. The contributions of the chemical constituents of the
plant to the bioactivities were not clearly correlated and
identified. It is necessary for the immunomodulatory activity
studies of the plant extracts to be accompanied with analyses of
their bioactive compounds and identification of the chemical
markers for standardization purposes. The extracts used in these
studies should be quantitative and qualitative analyzed by using
validated analytical methods. Some of the bioactive compounds
especially the curcuminoids (curcumin, demethoxycurcumin and
bisdemethoxycurcumin) and some sesquiterpenoids have been
isolated from the extracts and their mechanistic effects in
modulating the immune system have been determined.
However, more mechanistic studies should be carried out for
in depth understanding of the modulating effects of the plant
samples on the innate and adaptive immune system. Of all the
Curcuma species investigated, the immunomodulatory effects of
C. longa and its major compound, curcumin, were the most
studied. Their modulatory effects on various signaling pathways
at molecular level have been reported. However, extensive
molecular work on the other Curcuma species need to be
carried out. Despite the regulatory requirements for clinical
studies and sufficient data have not been generated on
preclinical testing, there were already reports on a few
unsystematic case studies to evaluate the immunomodulating
properties of Curcuma species in human. Some clinical trials have
been conducted on C. zanthorrhiza but they were unsystematic
and not well designed. There was also lack of sufficient preclinical
data and the extracts were not appropriately standardized. For
clinical studies, sufficient preclinical testing should be generated
using standardized extracts, which include bioavailability,
pharmacokinetic and toxicological studies, before they can be
subjected to clinical studies. Of all the bioactive metabolites of
Curcuma species, only curcumin is undergoing extensive clinical
trials for its anti-inflammatory properties and potential use as an
adjuvant in the treatment of cancer. Curcumin has showed
significant ability to modulate the immune response in
experimental and clinical studies. However, more systematic
and operationally thorough controlled randomized trials are
needed to prove its safety and efficacy for human use. Other
compounds from Curcuma species such as xanthorrhizol,
curdione, curcuzedoalide, isoprocurcumenol and turmeronols
have also been reported to modulate various lineages of
immune response. More in depth studies including elaborate
bioavailability, preclinical pharmacokinetics and toxicity studies
are required to understand the underlying mechanisms and safety
level before clinical trials can be pursued for development into
potent and safe immunomodulatory agents.
AUTHOR CONTRIBUTIONS
All authors participated in the concept and preparation of draft,
revised the manuscript, and approved the final version for
submission to this journal.
FUNDING
This work was funded by the Ministry of Education and Culture
Republic of Indonesia under the World Class University
Program Year 2020. The grant number is 1879/UN5.1.R/SK/
PPM/2020.
REFERENCES
Abas, F., Lajis, N. H., Israf, D. A., Khozirah, S., and Umi Kalsom, Y. (2006).
Antioxidant and Nitric Oxide Inhibition Activities of Selected Malay
Traditional Vegetables. Food Chem. 95, 566–573. doi:10.1016/j.foodchem.
2005.01.034
Adeshina, I., Aewale, Y. A., and Tiamiyu, L. O. (2017). Growth Performance and
Innate Immune Response of Clarias gariepinus Infected with Aeromonas
hydrophila Fed Diets Fortified with Curcuma longa Leaf. West. Afr. J. Appl.
Ecol. 25, 87–99.
Afolayan,F.I.D.,Erinwusi,B.,andOyeyemi,O.T.(2018).ImmunomodulatoryActivity
of Curcumin-Entrapped poly d,l -lactic- Co -glycolic Acid Nanoparticles in Mice.
Integr.Med.Res.7, 168–175. doi:10.1016/j.imr.2018.02.004
Alambra,J.R.,Alenton,R.R.R.,Gulpeo,P.C.R.,Mecenas,C.L.,Miranda,
A. P., Thomas, R. C., et al. (2012). Immunomodulatory Effects of
Turmeric, Curcuma longa (Magnoliophyta, Zingiberaceae) on
Macrobrachium rosenbergii (Crustacea, Palaemonidae) against Vibrio
alginolyticus (Proteobacteria, Vibrionaceae). Aquac. Aquar. Conserv
Legis. 5, 13–17.
Alsahli, A. A., Alaraidh, I. A., Rashad, Y. M., and Abdel razik, E. S. (2018). Extract
from Curcuma longa L. Triggers the Sunflower Immune System and Induces
Defence-Related Genes against Fusarium Root Rot. Phytopathol. Mediterr. 57,
26–36. doi:10.14601/Phytopathol_Mediterr-21176
Anand, P., Kunnumakkara, A. B., Newman, R. A., and Aggarwal, B. B. (2007).
Bioavailability of Curcumin: Problems and Promises. Mol. Pharmaceutics 4,
807–818. doi:10.1021/mp700113r
Anggriani, L., Yasmin, A., Wulandari, A. R., Leksono, G. M., Ikawati, M., and
Meiyanto, E. (2019). Extract of Temu Ireng (Curcuma aeruginosa Roxb.)
Rhizome Reduces Doxorubicin-Induced Immunosuppressive Effects. AIP
Conf. Proc. 2099, 020001. doi:10.1063/1.5098406
Anisuzzama, M., Sharmin, S. A., Mondal, S. C., Sultana, R., Khalekuzza, M., Alam,
I., et al. (2008). In vitro Microrhizome Induction in Curcuma zedoaria
(Christm.) Roscoe-A Conservation Prioritized Medicinal Plant. J. Biol. Sci.
8, 1216–1220. doi:10.3923/jbs.2008.1216.1220
Arifin, P. F., Setiawan, I. M., PurwantiningsihAstuti, R. B., Azima, F., Susilowidodo,
R. A., et al. (2020). Acute Toxicity Evaluation of Temulawak (Curcuma
xanthorrhiza Roxb.) Hepatoprotective Supplement. Res. J. Pharm. 11 (2),
13–18. doi:10.33887/rjpbcs/2020.11.2.3
Aristyani, S., Widyarti, S., and Sumitro, S. B. (2018). Network Analysis of
Indigenous Indonesia Medical Plants for Treating Tuberculosis.
Pharmacogn. J. 10, 1159–1164. doi:10.5530/pj.2018.6.198
Arshad, L., Md. Areeful Haque, Md. A., Bukhari, S. N. A., and Jantan, I. (2017). An
Overview of Structure-Activity Relationship Studies of Curcumin Analogs as
Antioxidant and Anti-Inflammatory Agents. Future Med. Chem. 9, 605–626.
doi:10.4155/fmc-2016-0223
Artfire (2016). Rare Curcuma amada. Available at: https://www.artfire.com/
(Accessed December 1, 2020).
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311921
Yuandani et al. Immunomodulatory Effects of Curcuma Species

Arunkumar, P., Ramasubramanian, V., and Munirasu, S. (2016). Effect of Curcuma
longa Enriched Mesocylops Thermocyclopoides on Fresh Water Fish, Cyprinus
carpio.Int. J. Res. Dev. Pharm. L. Sci. 6, 2848–2492.
Astana,P.R.W.,Ardiyanto,D.,andMana,T.A.(2018).Changeinqualityoflifeand
CD4+ value on HIV/AIDS patients with immuostimulant Jamu Formula in Sragen
regency. Indonesian J. Clin. Pharm. 7, 227-235. doi:10.15416/ijcp.2018.7.4.227
Ayati, Z., Ramezani, M., Amiri, M. S., Moghadam, A. T., Rahimi, H., Abdollahzade,
A., et al. (2019). Ethnobotany, Phytochemistry and Traditional Uses of
Curcuma spp. and Pharmacological Profile of Two Important Species (C.
longa and C. zedoaria): A Review. Cpd 25, 871–935. doi:10.2174/
1381612825666190402163940
Ayodele, V. O., Olowe, O. M., Afolabi, C. G., and Kehinde, I. A. (2018).
Identification, Assessment of Diseases and Agronomic Parameters of
Curcuma amada Roxb (Mango ginger). Curr. Plant Biol. 15, 51–57. doi:10.
1016/j.cpb.2018.10.001
Balaji, S., and Chempakam, B. (2010). Toxicity Prediction of Compounds from
Turmeric (Curcuma longa L). Food Chem. Toxicol. 48 (10), 2951–2959. doi:10.
1016/j.fct.2010.07.032
Baroroh, H. N., Ikawati, Z., and Sudarman, K. (2011). A Safety Study of Extract
Combination of Legundi (Vitex trifolia L.) Leaves and Temulawak (Curcuma
xanthorrhiza R.) Rhizome as Anti-allergy in Healthy Volunteers. Int. J. Pharm.
Pract. 2, 165–170.
Beutler, B. (2004). Innate Immunity: An Overview. Mol. Immunol. 40, 845–859.
doi:10.1016/j.molimm.2003.10.005
Beyaert, R., Beaugerie, L., Van Assche, G., BrochezRenauld, L. J. C., Renauld, J.-C.,
Viguier, M., et al. (2013). Cancer Risk in Immune-Mediated Inflammatory
Diseases (IMID). Mol. Cancer 12, 98. doi:10.1186/1476-4598-12-98
Bogdan, C. (2001). Nitric Oxide and the Immune Response. Nat. Immunol. 2,
907–916. doi:10.1038/ni1001-907
Carvalho, F., Vassão, R., Nicoletti, M., and Maria, D. (2010). Effect of Curcuma
zedoaria Crude Extract against Tumor Progression and Immunomodulation.
J. Venom. Anim. Toxins Incl. Trop. Dis. 16, 324–341. doi:10.1590/s1678-
91992010000200013
Castro, C. N., Barcala Tabarrozzi, A. E., Winnewisser, J., Gimeno, M. L., Antunica
Noguerol, M., Liberman, A. C., et al. (2014). Curcumin Ameliorates
Autoimmune Diabetes. Evidence in Accelerated Murine Models of Type 1
Diabetes. Clin. Exp. Immunol. 177, 149–160. doi:10.1111/cei.12322
Chanda, S., and Ramachandra, T. V. (2019). Phytochemical and Pharmacological
Importance of Turmeric (Curcuma longa): A Review. Res. Rev. J. Pharm. 9,
16–23.
Chandrasekaran, C. V., Sundarajan, K., Kannan, J. R., Gururaja, G. M.,
Mundkinajeddu, D., and Agarwal, A. (2013). Immune-Stimulatory and
Anti-inflammatory Activities of Curcuma longa Extract and its
Polysaccharide Fraction. Pharmacognosy Res. 5, 71–79. doi:10.4103/0974-
8490.110527
Chapel, H., Haeney, M., Misbah, S., and Snowden, N. (2006). Clinical Immunology.
5th Edn. Massachusetts: Blackwell Publishing.
Choudhury, D., Ghosal, M., Das, A. P., and Mandal, P. (2013). Development of
Single Node Cutting Propagation Techniques and Evaluation of Antioxidant
Activity of Curcuma aeruginosa Roxnurgh Rhizome. Int. J. Pharm.Sci. Res. 5,
227–234.
Cianciulli, A., Calvello, R., Porro, C., Trotta, T., Salvatore, R., and Panaro, M. A.
(2016). PI3k/Akt Signalling Pathway Plays a Crucial Role in the Anti-
inflammatory Effects of Curcumin in LPS-Activated Microglia. Int.
Immunopharmacol. 36, 282–290. doi:10.1016/j.intimp.2016.05.007
Devaraj, S., Esfahani, A. S., Ismail, S., Ramanathan, S., and Yam, M. F.
(2010). Evaluation of the Antinociceptive Activity and Acute Oral
Toxicity of Standardized Ethanolic Extract of the Rhizome of Curcuma
xanthorrhiza Roxb. Molecules 15 (4), 2925–2934. doi:10.3390/
molecules15042925
Dewi, M., Aries, M. H., Dwiriani, C. M., and Januwati, N. (2012). Knowledge on
Health Benefit of Curcuma and the Clinical Trial of Its Effect on Humoral
Immune System in Obese Adults. Indones. J. Agric. 17, 166–171.
Dosoky, N. S., and Setzer, W. N. (2018). Chemical Composition and Abiological
Activities of Essential Oils of Curcuma Species. Nutrients 10, 1196. doi:10.3390/
nu10091196
Drugbank (2021). Curcuma xanthorrhiza oil. Available at: https://go.drugbank.
com/drugs/DB11265 (Accessed January 26, 2021).
Dutta, B. (2015). Study of Secondary Metabolite Constituents and Curcumin Contents
of Six Different Species of Genus Curcuma.J. Med. Plants Stud. 3, 116–119.
Economou, M., Zambeli, E., and Michopoulos, S. (2009). Incidence and Prevalence
of Crohn’s Disease and its Etiological Influences. Ann. Gastroenterol. 22,
158–167.
Elgert, K. D. (2009). Immunology Understanding the Immune System. 2nd Edn.
Virginia: A John Wiley & Sons, Inc., Publication.
Ervintari, F., Puspitawati, R., and Utami, S. (2019). Effect of Curcuma Xanthorrhiza
Roxb. Ethanol Extract on the Viability of Streptococcus Mutans and
Streptococcus Sanguinis Dual-Species Biofilms. Int. J. App. Pharm. 11,
75–78. doi:10.22159/ijap.2019.v11s1.181
Faradilla, M., and Iwo, M. I. (2014). Immunomodulatory Effect of Polysaccharide
from White Turmeric [Curcuma zedoria (Christm.) Roscoe)]. Indonesian
J. Pharm. 12, 73–278.
Firtel, R. A., and Chung, C. Y. (2000). The Molecular Genetics of Chemotaxis:
Sensing and Responding to Chemoattractant Gradients. BioEssays 22, 603–615.
doi:10.1002/1521-1878(200007)22:7<603::aid-bies3>3.0.co;2-#
Flores, G. (2017). Curcuma longa L. Extract Improves the Cortical Neural
Connectivity during the Aging Process. Neural Regen. Res. 12, 875–880.
doi:10.4103/1673-5374.208542
Fu, Y., Gao, R., Cao, Y., Guo, M., Wei, Z., Zhou, E., et al. (2014). Curcumin
Attenuates Inflammatory Responses by Suppressing TLR4-Mediated NF-κB
Signaling Pathway in Lipopolysaccharide-Induced Mastitis in Mice. Int.
Immunopharmacol. 20, 54–58. doi:10.1016/j.intimp.2014.01.024
Fürst, R., and Zündorf, I. (2014). Plant-Derived Anti-Inflammatory Compounds:
Hopes and Disappointments Regarding the Translation of Preclinical
Knowledge into Clinical Progress. Mediators Inflamm. 2014, 146832. doi:10.
1155/2014/146832
Gao, X., Kuo, J., Jiang, H., Deeb, D., Liu, Y., Divine, G., et al. (2004).
Immunomodulatory Activity of Curcumin: Suppression of Lymphocyte
Proliferation, Development of Cell-Mediated Cytotoxicity, and Cytokine
Production in vitro.Biochem. Pharmacol. 68, 51–61. doi:10.1016/j.bcp.2004.
03.015
Geetha, S., Singh, V., Ram, M. S., Ilavazhagan, G., Banerjee, P. K., and Sawhney, R.
C. (2005). Immunomodulatory Effects of Seabuckthorn (Hippophae
rhamnoides L.) against Chromium (VI) Induced Immunosuppression. Mol.
Cel. Biochem. 278, 101–109. doi:10.1007/s11010-005-7095-9
George, M., and Britto, S. J. (2015). Phytochemicaland Antioxidant Studies on the
Essential Oil of the Rhizome of Curcuma Aeruginosa Roxb. Int. Res. J. Pharm. 6,
573–579. doi:10.7897/2230-8407.068113
Guimarães,A.F.,Vinhas,A.C.A.,Gomes,A.F.,Souza,L.H.,andKrepskya,P.B.
(2020). Essential Oil of Curcuma longa L. Rhizomes Chemical Composition, Yield
Variation and Stability. Quim. Nova 43, 909–913. doi:10.21577/0100-4042.20170547
Hardiwati, K. T., Seninha, M., Lay, B. W., and Yanti (2019). Curcuminoid Cider
Fermented from Curcuma xanthorrhiza Curcuminoids Attenuates Gene
Expression Related to Obesity-Induced Inflammation in
Hypercholesterolaemic Rats. Int. Food Res. J. 26, 859–867.
Harun, N. H., Septama, A. W., and Jantan, I. (2015). Immunomodulatory Effects of
Selected Malaysian Plants on the CD18/11a Expression and Phagocytosis
Activities of Leukocytes. Asian Pac. J. Trop. Biomed. 5, 48–53. doi:10.1016/
s2221-1691(15)30170-2
Hassan, A. A. M., Yacout, M. H., Khalel, M. S., Hafsa, S. H. A., Ibrahim, M. A. R.,
Mocuta, D. N., et al. (2018). Effects of Some Herbal Plant Supplements on
Growth Performance and the Immune Response in Nile Tilapia (Oreochromis
Niloticus). Sciendo 1, 134–141. doi:10.2478/alife-2018-0020
Hong, G. W., Hong, S. L., Lee, G. S., Yaacob, H., and Malek, S. N. A. (2016). Non-
Aqueous Extracts of Curcuma mangga Rhizomes Induced Cell Death in Human
Colorectal Adenocarcinoma Cell Line (HT29) via Induction of Apoptosis and
Cell Cycle Arrest at G0/G1 Phase. Asian Pac. J. Trop. Med. 9, 8–18. doi:10.1016/
j.apjtm.2015.12.003
HMPC (2017). Assessment report on Curcuma longa L. (C. domestica Valeton)
rhizoma. London: European Medicines Agency, 1–5.
HMPC (2014). Assessment report on Curcuma xanthorrhiza Roxb. (C.
xanthorrhiza D. Dietrich), rhizoma. London: European Medicines
Agency, 15.
Huang, J. K., Kim, A. J., Sohn, J. H., Han, K. L., and Choo, J. H. (2010).
Immunostimulating Polysaccharide Isolated from Curcuma xanthorrhiza
and Manufacturing Method Thereof. Patent Appl. Publ.,1–11.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311922
Yuandani et al. Immunomodulatory Effects of Curcuma Species

Huang, Y., Xue, C., He, W., and Zhao, X. (2019). Inhibition Effect of Zedoary
Turmeric Oil on Listeria Monocytogenes and Staphylococcus aureus Growth and
Exotoxin Proteins Production. J. Med. Microbiol. 68, 657–666. doi:10.1099/
jmm.0.000949
Ikawati, Z., Hertiani, T., and Izzati, F. (2019). Immunomodulatory Activity of an
Indonesian Herbal Formulation for Respiratory Disorder. Phcog Mag. 15,
130–134. doi:10.4103/pm.pm_314_18
Ilene, A., Sandhika, W., and Hasanatuludhhiyah, N. (2020). Effect of Javanese
Turmeric (Curcuma xanthorrhiza) Extract on Hepatitis Model of Alcohol-
Induced Mice. Jurnal Kedokteran Brawijaya 1, 39–44. doi:10.21776/ub.jkb.
2020.031.01.8
Inzaugarat, M. E., Matteo, E. D., Baz, P., Lucero, D., Garcia, C. C., Ballerga, E. G.,
et al. (2017). New Evidence for the Therapeutic Potential of Curcumin to Treat
Nonalcoholic Faty Liver Disease in Humans. Plos One 12 (3), e0172900. doi:10.
1371/journal.pone.0172900
Jain, S. K., Rains, J., Croad, J., Larson, B., and Jones, K. (2009). Curcumin
Supplementation Lowers TNF-α, IL-6, IL-8, and MCP-1 Secretion in High
Glucose-Treated Cultured Monocytes and Blood Levels of TNF-α, IL-6, MCP-
1, Glucose, and Glycosylated Hemoglobin in Diabetic Rats. Antioxid. Redox
Signaling 11, 241–249. doi:10.1089/ars.2008.2140
Jang, M. K., Lee, H. J., Kim, J. S., and Ryu, J.-H. (2004). A Curcuminoid and Two
Sesquiterpenoids from Curcuma Zedoaria as Inhibitors of Nitric Oxide
Synthesis in Activated Macrophages. Arch. Pharm. Res. 27, 1220–1225.
doi:10.1007/bf02975885
Jang, M., Sohn, D., and Ryu, J.-H. (2001). A Curcuminoid and Sesquiterpenes as
Inhibitors of Macrophage TNF-αRelease from Curcuma Zedoaria. Planta Med.
67, 550–552. doi:10.1055/s-2001-16482
Jantan, I., Saputri, F. C., Qaisar, M. N., and Buang, F. (2012). Correlation between
Chemical Composition of Curcuma domestica and Curcuma xanthorrhiza and
Their Antioxidant Effect on Human Low-Density Lipoprotein Oxidation. Evid.
Based Complement. Alternat. Med. 2012, 438356. doi:10.1155/2012/438356
Jantan, I., Harun, N. H., Septama, A. W., Murad, S., and Mesaik, M. A. (2011).
Inhibition of Chemiluminescence and Chemotactic Activity of Phagocytes
in vitro by the Extracts of Selected Medicinal Plants. J. Nat. Med. 65,
400–405. doi:10.1007/s11418-010-0492-8
Jatoi, S. A., Kikuchi, A., Gilani, S. A., and Watanabe, K. N. (2007). Phytochemical,
Pharmacological and Ethnobotanical Studies in Mango Ginger (Curcuma
amada Roxb.; Zingiberaceae). Phytother. Res. 21, 507–516. doi:10.1002/ptr.
2137
Jose, S., and Thomas, T. D. (2014). Comparative Phytochemical and Anti-Bacterial
Studies of Two Indigenous Medicinal Plants Curcuma caesia Roxb. and
Curcuma aeruginosa Roxb. Int. J. Green. Pharm. 8, 65–71. doi:10.4103/
0973-8258.126828
Jurenka, J. S. (2009). Anti-Inflammatory Properties of Curcumin, a Major
Constituent of Curcuma longa: A Review of Preclinical and Clinical
Research. Altern. Med. Rev. 14, 141–153.
Kaewkroek, K., Wattanapiromsakul, C., and Tewtrakul, S. (2010). Anti-
Inflammatory Mechanisms of Compounds from Curcuma mangga
Rhizomes using RAW264.7 Macrophage Cells. Nat. Prod. Commun. 5,
1547–1550. doi:10.1177/1934578x1000501006
Kaewkroek, K., Wattanapiromsakul, C., and Tewtrakul, S. (2009). Nitric Oxide
Inhibitory Substances from Curcuma mangga Rhizomes. Songklanakarin J. Sci.
Technol. 31, 293–297.
Kalim, H., Handono, K., Khalasha, T., Pratama, M., Dantara, T. I., Wulandari, A.,
et al. (2017). Immune Modulation Effects of Curcumin in Pristane-Induced
Lupus Mice. Indian J. Rheumatol. 12, 86–93. doi:10.4103/injr.injr_95_16
Kaliyadasa, E., and Samarasinghe, B. A. (2019). A Review on Golden Species of
Zingiberaceae Family Around the World: Genus Curcuma.Afr. J. Agric. Res. 14,
519–531. doi:10.5897/AJAR2018.13755
Karchuli, M. S., and Pradhan, D. (2011). Curcuma amada Roxb. Rhizome Extract
Modulates Cellular and Humoral Immune System. Pharmcologyonline 3,
947–952.
Kavitha, R., and Mahadevi, R. (2020). Phytochemical and Pharmacological
Properties of Curcuma amada: A Review. Int. J.Pharm. Sci. Res. 11,
3546–3555. doi:10.26452/ijrps.v11i3.2510
Kim, A.-J., Kim, Y.-O., Shim, J.-S., and Hwang, J.-K. (2007). Immunostimulating
Activity of Crude Polysaccharide Extract Isolated from Curcuma xanthorrhiza
Roxb. Biosci. Biotechnol. Biochem. 71, 1428–1438. doi:10.1271/bbb.60241
Kim, G.-Y., Kim, K.-H., Lee, S.-H., Yoon, M.-S., Lee, H.-J., Moon, D.-O., et al.
(2005). Curcumin Inhibits Immunostimulatory Function of Dendritic Cells:
MAPKs and Translocation of NF-κB as Potential Targets. J. Immunol. 174,
8116–8124. doi:10.4049/jimmunol.174.12.8116
Kim, K. I., Shin, K. S., Jun, W. J., Hong, B. S., Shin, D. H., Cho, H. Y., et al. (2001).
Effects of Polysaccharides from Rhizomes of Curcuma Zedoaria on
Macrophage Functions. Biosci. Biotechnol. Biochem. 65, 2369–2377. doi:10.
1271/bbb.65.2369
Kim, M.-B., Kim, C., Song, Y., and Hwang, J.-K. (2014). Antihyperglycemic and
Anti-Inflammatory Effects of Standardized Curcuma xanthorrhiza Roxb.
Extract and Its Active Compound Xanthorrhizol in High-Fat Diet-Induced
Obese Mice. Evid. Based. Complement. Alternat. Med. 2014, 205915. doi:10.
1155/2014/205915
Kim, O. K., Yoo, S. A., Nam, D.-E., Kim, Y., Kim, E., Jun, W., et al. (2014).
Immunomodulatory Effects of Curcuma longa L. Extract in LP-BM5 Murine
Leukemia Viruses-Induced Murine Acquired Immune Deficiency Syndrome.
J. Korean Soc. Food Sci. Nutr. 43, 1317–1324. doi:10.3746/jkfn.2014.43.9.1317
Kim, S., Kook, K. E., Kim, C., and Hwang, J.-K. (2018). Inhibitory Effects of
Curcuma xanthorrhiza Supercritical Extract and Xanthorrhizol on LPS-
Induced Inflammation in HGF-1 Cells and RANKL-Induced
Osteoclastogenesis in RAW264.7 Cells. J. Microbiol. Biotechnol. 28,
1270–1281. doi:10.4014/jmb.1803.03045
Kobayashi, S. D., Voyich, J. M., Burlak, C., and DeLeo, F. R. (2005). Neutrophils in
the Innate Immune Response. Arch. Immunol. Ther. Exp. 53, 505–517.
Kumar, S., Ahuja, V., Sankar, M. J., Kumar, A., and Moss, A. C. (2012). Curcumin
for Maintenance of Remission in Ulcerative Colitis. Cochrane Database Syst.
Rev. 10, CD008424. doi:10.1002/14651858.CD008424.pub2
Kumolosasi, E., Ibrahim, S. N. A., Ahmad Shukri, S. M., and Ahmad, W. (2018).
Immunostimulant Activity of Standardised Extracts of Mangifera Indica Leaf
and Curcuma Domestica Rhizome in Mice. Trop. J. Pharm. Res. 17, 77–84.
doi:10.4314/tjpr.v17i1.12
Kusumarin, N., Lianahand Hafsha, M. (2020). Evaluation of the Potential of Ireng
Gathering Starch (Curcuma aeruginosa Roxb.) as Alternative Food Ingredients
and the Processed Organoleptic Aspects. Int. J. Agric. Biol. Sci. 2, 71–76. doi:10.
5281/zenodo.3710771
Lakshmi, S., Padmaja, G., and Remani, P. (2011). Antitumour Effects of
Isocurcumenol Isolated from Curcuma zedoaria Rhizomes on Human and
Murine Cancer Cells. Int. J. Med. Chem. 2011, 253962. doi:10.1155/2011/
253962
Lee, T. K., Trinh, T. A., Lee, S. R., Kim, S., So, H. M., Moon, E., et al. (2019).
Bioactivity-Based Analysis and Chemical Characterization of Anti-
Inflammatory Compounds from Curcuma zedoaria Rhizomes using LPS-
Stimulated RAW264.7 Cells. Bioorg. Chem. 82, 26–32. doi:10.1016/j.bioorg.
2018.09.027
Lestari, M. D., Arief, M., and Satyantini, W. H. (2019). Addition of Curcuma
(Curcuma xanthorrhiza) as an Antioxidant on African Catfish (Clarias
gariepinus) Commercial Fish Feeding. Int. J. Civ. Eng. 10, 380–385.
Li, S., Yuan, W., Deng, G., Wang, P., Yang, P., and Aggarwal, B. B. (2011). Chemical
Composition and Product Quality Control of Turmeric (Curcuma longa L.).
Topharmcj 5, 28–54. doi:10.2174/2210290601102010028
Liju, V. B., Jeena, K., and Kuttan, R. (2013). Acute and Subchronic Toxicity as Well
as Mutagenic Evaluation of Essential Oil from Turmeric (Curcuma longa L).
Food Chem. Toxicol. 53, 52–61. doi:10.1016/j.fct.2012.11.027
Listyawati, S. (2006). Toxicity Studies of the Rhizome Curcuma xanthorrhiza Roxb.
and Curcuma zedoaria Roscoe on Hematological and Male Reproduction
System of Mice (Mus musculus L.). J. Nat. Prod. Biochem. 4 (1), 10–13.
doi:10.13057/biofar/f040103
Liu,Y.,andNair,M.G.(2011).LabdaneDiterpenesinCurcuma mangga
Rhizomes Inhibit Lipid Peroxidation, Cyclooxygenase Enzymes and
Human Tumour Cell Proliferation. Food Chem. 124, 527–532. doi:10.
1016/j.foodchem.2010.06.064
Lobo, R., Prabhu, K. S., Shirwaikar, A., and Shirwaikar, A. (2009). Curcuma
zedoaria Rosc. (white turmeric): A Review of Its Chemical, Pharmacological
and Ethnomedicinal Properties. J. Pharm. Pharmacol. 61, 13–21. doi:10.1211/
jpp/61.01.0003
Luckheeram, R. V., Zhou, R., Verma, A. D., and Xia, B. (2012). CD4+T Cells:
Differentiation and Functions. Clin. Develop. Immunol. 2012, 1. doi:10.1155/
2012/925135
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311923
Yuandani et al. Immunomodulatory Effects of Curcuma Species

Luster, A. D. (2001). “Chemotaxis: Role in Immune Response,”in Encyclopedia of
Life Sciences. Massachusetts: Nature Publishing Grup, 1–12.
Malek, S. N. A., Lee, G. S., Hong, S. L., Yaacob, H., Wahab, N. A., Faizal Weber, J.-
F., et al. (2011). Phytochemical and Cytotoxic Investigations of Curcuma
mangga Rhizomes. Molecules 16, 4539–4548. doi:10.3390/molecules16064539
Mary, H. P., Susheela, G. K., Jayasree, S., Nizzy, A., Rajagopal, B., and Jeeva, S.
(2012). Phytochemical Characterization and Antimicrobial Activity of
Curcuma xanthorrhiza Roxb. Asian Pac. J. Trop. Biomed. 2, S637–S640.
doi:10.1016/s2221-1691(12)60288-3
Matsuda, H., Tewtrakul, S., Morikawa, T., Nakamura, A., and Yoshikawa, M.
(2004). Anti-allergic Principles from Thai Zedoary: Structural Requirements of
Curcuminoids for Inhibition of Degranulation and Effect on the Release of
TNF-αand IL-4 in RBL-2H3 Cells. Bioorg. Med. Chem. 12, 5891–5898. doi:10.
1016/j.bmc.2004.08.027
Mauren, F. M., Yantiand Lay, B. W. (2016). Efficacy of Oral Curcuminoid Fraction
from Curcuma xanthorrhiza and Curcuminoid Cider in High-Cholesterol Fed
Rats. Pharmacogn. Res. 8, 153–159. doi:10.4103/0974-8490.181468
Miksusanti (2012). Lymphocyte Proliferation by Temu Lawak (Curcuma
xanthorrhiza ROXB) Essential Oil. Int. Conf. Bioinform. Biomed. Eng. 29,
205–209.
Mooraki, N., Batmany, Y., Zoriehzahra, S. J., and Kakoolaki, S. (2019). Evaluating
the Effect of using Turmeric (Curcuma longa) of Growth Performance and
Hematological Parameters of the Ornamental Fish, Green Terror (Andinocara
rivulatus). J. Surv. Fish. Sci. 5, 37–47. doi:10.18331/sfs2019.5.2.5
Morales, G. A., Noratto, G., and Talcott, S. U. M. (2012). Standardized
Curcuminoid Extract (Curcuma longa l.) Decreases Gene Expression Related
to Inflammation and Interacts with Associated microRNAs in Human
Umbilical Vein Endothelial Cells (HUVEC). Food Funct. 3, 1286–1293.
doi:10.1039/c2fo30023k
Naderi, M., Akbari, M. R., Asadi, K. E., Khaksar, K., and Khajali, F. (2014). Effects
of Dietary Inclusion of Turmeric (Curcuma longa) and Cinnamon
(Cinnamomum verum) Powders on Performance, Organs Relative Weight
and Some Immune System Parameters in Broiler Chickens. Poult. Sci. J. 2,
153–163. doi:10.22069/PSJ.2014.1963
Nan, F. H., Agus, P. A. S., MargieBargir, B., and Lee, M. C. (2014). The Effects of
Curcuma zedoaria and Zingiber zerumbet on Non-Specific Immune Responses
of Grouper Epinephelus coioides.Iran. J. Fish. Sci. 14, 598–611.
Okamoto, A., Noble, E. E., Tyagi, E., Ying, Z., Zhuang, Y., and Gomez-Pinilla, F.
(2015). Curcumin Boosts DHA in the Brain: Implications for the Prevention of
Anxiety Disorders. Biochim. Biophys. Acta (BBA) - Mol. Basis Dis. 1852,
951–961. doi:10.1016/j.bbadis.2014.12.005
Okuda-Hanafusa, C., Uchio, R., Fuwa, A., Kawasaki, K., Muroyama, K.,
Yamamoto, Y., et al. (2019). Turmeronol A and Turmeronol B from
Curcuma longa Prevent Inflammatory Mediator Production by
Lipopolysaccharide-Stimulated RAW264.7 Macrophages, Partially via
Reduced NF-κB Signaling. Food Funct. 10, 5779–5788. doi:10.1039/c9fo00336c
Oon, S. F., Nallappan, M., Tee, T. T., Shohaimi, S., Kassim, N. K., Sa’ariwijaya, M. S.
F., et al. (2015). Xanthorrhizol: A Review of its Pharmacological Activities and
Anticancer Properties. Cancer Cel. Int. 15, 1–15. doi:10.1186/s12935-015-
0255-4
Palpu, P., Rao, C. V., Rawat, A. K. S., Pjha, S. K., and Reddy, G. D. (2008). Anti-
Allergic Herbal Formulation. U.S. Patent No 7344739 B2. Washington, DC: U.S.
Patent and Trademark Office.
Pan, M. H., Wu, J. C., Ho, C. T., and Badmaev, V. (2017). Effect of Water Extract of
Curcuma longa (L.) Roots on Immunity and Telomerase Function.
J. Complement. Integr. Med. 14, 1–6. doi:10.1515/jcim-2015-0107
Park, S.-J., Lee, D., Lee, M., Kwon, H.-O., Kim, H., Park, J., et al. (2018). The Effects
of Curcuma longa L., Purple Sweet Potato, and Mixtures of the Two on
Immunomodulation in C57BL/6J Mice Infected with LP-BM5 Murine
Leukemia Retrovirus. J. Med. Food 21, 689–700. doi:10.1089/jmf.2017.4093
Patil, U. S., Jaydeokar, A. V., and Bandawane, D. D. (2012). Immunomodulators: A
Pharmacological Review. Int. J. Pharm. Sci. 4, 30–36.
Pawitan, J. A. (2020). Curcumin as Adjuvant Therapy in COVID-19: Friend or
Foe?. J.Int. Dent. Med. Res. 13, 824–829.
Pisani, L. F., Lecchi, C., Invernizzi, G., Sartorelli, P., Savoini, G., and Ceciliani, F.
(2009). In vitro Modulatory Effect of ω-3 Polyunsaturated Fatty Acid (EPA and
DHA) on Phagocytosis and ROS Production of Goat Neutrophils. Vet.
Immunol. Immunopathol. 131, 79–85. doi:10.1016/j.vetimm.2009.03.018
Policegoudra, R. S., Divakar, S., and Aradhya, S. M. (2006). Identification of
Difurocumenonol, a New Antimicrobial Compound from Mango Ginger
(Curcuma amada Roxb.) Rhizome. J. Appl. Microbiol. 102, 1594–1602.
doi:10.1111/j.1365-2672.2006.03186.x
Policegoudra, R. S., Aradhya, S. M., and Singh, L. (2011). Mango Ginger (Curcuma
amada Roxb.) - A Promising Spice for Phytochemicals and Biological Activities.
J. Biosci. 36, 739–748. doi:10.1007/s12038-011-9106-1
Purwanti, S., Agustina, L., Syamsu, J. A., Adriyansyah, A., and Latief, M. F. (2018).
Turmeric (Curcuma domestica) and Garlic (Allium sativum) towards Broiler
Immune System Infected by Salmonella pullorum Bacteria as a Feed Additive.
IOP Conf. Ser. Earth Environ. Sci. 247, 1–6. doi:10.1088/1755-1315/247/1/
012063
Putri, M. S. (2014). White Turmeric (Curcuma zedoaria): Its Chemical Substance
and the Pharmacological Benefit. J. Majority 3, 88–93.
Rahayu, D. U. C., Setyani, D. A., Dianhar, H., and Sugita, P. (2020). Phenolic
Compounds from Indonesian White Turmeric (Curcuma zedoaria) Rhizomes.
Asian J. Pharm. Clin. Res. 13, 194–198. doi:10.22159/ajpcr.2020.v13i7.38249
Rahim, N. A., Hassandarvish, P., Golbabapour, S., Ismail, S., Tayyab, S. , and
Abdulla, M. A. (2014). Gastroprotective Effect of Ethanolic Extract of
Curcuma xanthorrhiza Leaf against Ethanol-Induced Gastric Mucosal
Lesions in Sprague-Dawley Rats. Biomed. Res. Int. 2014, 416409. doi:10.
1155/2014/416409
Rajkumari, S., and Sanatombi, K. (2017). Nutritional Value, Phytochemical
Composition, and Biological Activities of Edible Curcuma Species: A
Review. Int. J. Food Prop. 20, S2668–S2687. doi:10.1080/10942912.2017.
1387556
Rattue, G. (2012). Autoimmune Disease Rates Increasing. Medical News Today.
Available at: http://www.medicalnewstoday.com/articles/246960.php
(Accessed February 15, 2015).
Ravikumar, N., and Naga Kavitha, C. (2020). Therapeutic Potential of Curcumin
on Immune Dysregulation in Comorbid Diabetic Asthma in Mice. Biomed.
Pharmacol. J. 13, 821–831. doi:10.13005/bpj/1948
Ruangsang, P., Tewtrakul, S., and Reanmongkol, W. (2010). Evaluation of the
Analgesic and Anti-Inflammatory Activities of Curcuma mangga Val and Zijp
Rhizomes. J. Nat. Med. 64, 36–41. doi:10.1007/s11418-009-0365-1
Saroj, P., Verma, M., Jha, K. K., and Pal, M. (2012). An Overview on
Immunomodulation. J. Adv. Sci. Res. 3, 7–12.
Sasikumar, B. (2005). Genetic Resources of Curcuma: Diversity, Characterization
and Utilization. Plant Genet. Resour. 3, 230–251. doi:10.1079/pgr200574
Sengupta, M., Sharma, G. D., and Chakraborty, B. (2011). Hepatoprotective and
Immunomodulatory Properties of Aqueous Extract of Curcuma longa in
Carbon Tetra Chloride Intoxicated Swiss Albino Mice. Asian Pac. J. Trop.
Biomed. 1, 193–199. doi:10.1016/s2221-1691(11)60026-9
Sethy, K., Swain, P., Behera, K., Sahoo, N., Agrawalla, J., Khandanga, S., et al.
(2017). Effect of Turmeric (Curcuma longa) Supplementation on Antioxidants
and Immunity of Broiler Birds. Livest. Sci. 8, 103–106. doi:10.4081/ijas.2015.
3870
Setiawati, M. C., Ikawati, Z., and Kertia, I. N. (2017). Antiinflamatory and
Antidepressive Activities of Extract Curcuma xanthorrhiza Roxb in Systemic
Lupus Erythematosus. Indonesian J. Pharm. 28, 185–190. doi:10.14499/
indonesianjpharm28iss3pp185
Setyati, W., Subagiyo, S., Pramesti, R., and Pramesti, D. (2019). Effectiveness of
Herbal Extract (Piper retrofractum,Curcuma aeruginosa,andCurcuma
zanthorrhiza) as Immunomodulator in Non-Specific Immunity System of
Tiger Grouper (Epinephelus fuscoguttatus) against Infection from Vibrio
Alginolyticus and Vibrio Parah. Sci. Technol. Indones. 4, 94–100. doi:10.
26554/sti.2019.4.4.94-100
Shabana, M. H., Shahy, E. M., Taha, M. M., Mahdy, G. M., and Mahmoud, M. H.
(2020). Phytoconstituents from Curcuma longa L. Aqueous Ethanol Extract and
Its Immunomodulatory Effect on Diabetic Infected Rats. Egypt. Pharmaceut J.
14, 36–43. doi:10.4103/1687-4315.154713
Shaikh, P. Z. (2011). Cytokines & Their Physiologic and Pharmacologic Functions
in Inflammation: A Review. Int. J. Pharm. Life Sci. 2, 1247–1263.
Sholikhah, E. N., Mustofa, M., Nugrahaningsih, D. A. A., Yuliani, F. S., Purwono,
S., Sugiyono, S., et al. (2020). Acute and Subchronic Oral Toxicity Study of
Polyherbal Formulation Containing Allium sativum L., Terminalia bellirica
(Gaertn.) Roxb., Curcuma aeruginosa Roxb., and Amomum compactum Sol. ex.
Maton in Rats. Biomed. Res. Int. 2020, 8609364. doi:10.1155/2020/8609364
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311924
Yuandani et al. Immunomodulatory Effects of Curcuma Species

Snapdeal (2020). Curcuma Amada. Available at: https://www.snapdeal.com/
(Accessed December 1, 2020).
Soleimani, V., Sahebkar, A., and Hosseinzadeh, H. (2018). Turmeric (Curcuma
longa) and its Major Constituent (Curcumin) as Nontoxic and Safe Substances:
Review. Phytotherapy Res. 32, 985–995. doi:10.1002/ptr.6054
Sudeepthi, N. L., Kumar, K. E., and Kola, P. K. (2014). Effect of Curcuma amada
(mango ginger) Roxb. n Scopolamine Induced Memory Deficit in Rats. Int.
J. Res. Pharm. Chem. 4, 1127–1134.
Sulfianti, A., Ningsih, S., and Agustini, K. (2019). Chemoprevention Effect of
Curcuma aeruginosa in DMBA-Induced Cytokines Production. Int. Res.
J. Pharm. 10, 54–59. doi:10.7897/2230-8407.100378
Sun, W., Wang, S., Zhao, W., Wu, C., Guo, S., Gao, H., et al. (2017). Chemical
Constituents and Biological Research on Plants in the Genus Curcuma.Crit.
Rev. Food Sci. Nutr. 57, 1451–1523. doi:10.1080/10408398.2016.1176554
Surh, Y.-J., Chun, K.-S., Cha, H.-H., Han, S. S., Keum, Y.-S., Park, K.-K., et al.
(2001). Molecular Mechanisms Underlying Chemopreventive Activities of
Anti-Inflammatory Phytochemicals: Down-Regulation of COX-2 and iNOS
through Suppression of NF-κB Activation. Mutat. Res./Fundam. Mol. Mech.
Mutagen. 480–481, 243–268. doi:10.1016/S0027-5107(01)00183-X
Tan, B., and Vanitha, J. (2004). Immunomodulatory and Antimicrobial Effects of
Some Traditional Chinese Medicinal Herbs: A Review. Cmc 11, 1423–1430.
doi:10.2174/0929867043365161
Thokchom, S. S., and Phucho, I. T. (2015). Elemental Analysis, Ditermination of
Alkaloid, Saponin and Flavonoid of Three Selected Species of Zingiberaceae
Family. Int. J. Pharm. Sci. Res. 6, 3044–3048. doi:10.13040/IJPSR.0975-8232.
6(7).3044-48
Tung, B. T., Nham, D. T., Hai, N. T., and Thu, D. K. (2019). Curcuma longa, the
Polyphenolic Curcumin Compound and Pharmacological Effects on Liver.
Dietary Interventions Liver Dis. 2019, 125–134. doi:10.1016/b978-0-12-814466-
4.00010-0
Uchio, R., Higashi, Y., Kohama, Y., Kawasaki, K., Hirao, T., Muroyama, K., et al.
(2017). A Hot Water Extract of Turmeric (Curcuma longa) Suppresses Acute
Ethanol-Induced Liver Injury in Mice by Inhibiting Hepatic Oxidative Stress
and Inflammatory Cytokine Production. J. Nutr. Sci. 6, 1–9. doi:10.1017/jns.
2016.43
USDA (2021). Natural Resources Conservation Services. Plants Database.
Available at: https://plants.usda.gov/java/ClassificationServlet?
sourcedisplay&classidCURCU (Accessed January 25, 2021).
Wahono, C. S., Setyorini, C. D., Kalim, H., Nurdianaand Handono, K. (2017a).
Effect of Curcuma xanthorrhiza Supplementation on Systemic Lupus
Erythematosus Patients with Hypovitamin D which were Given Vitamin D3
towards Disease Activity (SLEDAI), IL-6, and TGF-β1 Serum. Int. J. Rheumatol.
2017, 7687053. doi:10.1155/2017/7687053
Wahono, C. S., Wahyuni, Z. D., and Kalim, H. (2017b). Effect of Curcuma
xanthorrhiza Suplementation in Vitamin D3 Administration towards
Proteinuria, Serum Anti-dsDNA and IL-17 Levels on Systemic Lupus
Erythematosus (sle) Patients with Hypovitamin. D Int. J. Clin. Rheumatol.
12, 121–129.
Warrington, R., Watson, W., Kim, H. L., and Antonetti, F. R. (2011). An
Introduction to Immunology and Immunopathology. All Asth Clin. Immun.
7, S1. doi:10.1186/1710-1492-7-s1-s1
Weidner, M. S., Peterson, M. J., and Jacobsen, N. (2001). Certain Diterpenes and
Extracts or Concentrates of Curcuma amada Containing Them for Use as
Medicaments. U.S. Patent No 6235287 B1. Washington, DC: U.S. Patent
and Trademark Office.
Win, N. N., Ito, T., Ngwe, H., Win, Y. Y., PremaOkamoto, Y., et al. (2017). Labdane
Diterpenoids from Curcuma amada Rhizomes Collected in Myanmar and their
Antiproliferative Activities. Fitoterapia 122, 34–39. doi:10.1016/j.fitote.2017.
08.006
Wu, A., Noble, E. E., Tyagi, E., Ying, Z., Zhuang, Y., Pinilla, F. G., et al. (2015).
Curcumin boosts DHA in the brain: implications for the prevention of anxiety
disorders. Biochim. Biophys. Acta 1852, 951–961. doi:10.1016/j.bbadis.2014.
12.005
Xia, X., Pan, Y., Zhang, W.-Y., Cheng, G., and Kong, L.-D. (2006). Ethanolic
Extracts from Curcuma longa Attenuates Behavioral, Immune, and
Neuroendocrine Alterations in a Rat Chronic Mild Stress Model. Biol.
Pharm. Bull. 29, 938–944. doi:10.1248/bpb.29.938
Yadav, R. P., and Tarun, G. (2017). Versality of Turmeric: a Review the Golden
Spice of Life. J. Pharmacogn. Phytochem. 6, 41–46.
Yadav, V. S., Mishra, K. P., Singh, D. P., Mehrotra, S., and Singh, V. K. (2005).
Immunomodulatory Effects of Curcumin. Immunopharmacol. Immunotoxicol.
27, 485–497. doi:10.1080/08923970500242244
Yoo, S. A., Kim, O. K., Nam, D.-E., Kim, Y., Baek, H., Jun, W., et al. (2014).
Immunomodulatory Effects of Fermented Curcuma longa L. Extracts on RAW
264.7 Cells. J. Korean Soc. Food Sci. Nutr. 43, 216–223. doi:10.3746/jkfn.2014.
43.2.216
Yoo, S. A., Kim, O. K., Nam, D. E., Park, S. J., Han, D. K., Kwon, H. O., et al. (2013).
Immunomodulatory Effects of Curcuma longa L., Mulberry Leaves, and Purple
Sweet Potato Extracts: Modulation of Immune Functions during Murine
Leukemia Virus Infection. Nutrition 27, 1079. doi:10.1096/fasebj.27.
1_supplement.1079.18
Yu, Y., Shen, Q., Lai, Y., Park, S. Y., Qu, X., Lin, D., et al. (2018). Anti-Inflammatory
Effect of Curcumin in Microglial Cells. Front. Pharmacol. 9, 1–10. doi:10.3389/
fphar.2018.00386
YuandaniNugraha, S. E., Laila, L., Silaban, S. D., and Ramadhani, F. (2020).
Stimulatory Effect of Curcuma mangga on Immune Response against
Staphylococcus aureus.Nusantara Biosci. 12, 109–113. doi:10.13057/
nusbiosci/n120204
Yuandani, Y., and Suwarso, E. (2017a). Acute Toxicity of Ethanol Extract of
Curcuma mangga Rhizome. Asian J. Pharm. Clin. Res. 10, 383–385. doi:10.
22159/ajpcr.2017.v10i9.18398
Yuandani, Y., and Suwarso, E. (2017b). Immunomodulatory Effects of Ethanol
Extract of Curcuma mangga Rhizomes in Mice. Asian J. Pharm. Clin. Res. 10,
383–385. doi:10.22159/ajpcr.2017.v10i9.18398
YuandaniYuliasmi, S., Satria, D, F., Dongoran, R, S., Sinaga, M. H. A., and
Marpaung, N. (2019). Correlation between the Phytochemical Constituents
of Curcuma mangga and Its Immunomodulatory Effect. Rasayan J. Chem. 12,
1–6. doi:10.31788/rjc.2019.1215050
YuandaniYuliasmi, S., Yuliasmi, S., and Satria, D. (2018). Analysis of Compounds
and Immunostimulatory Properties of Curcuma mangga Rhizomes on Male
Mice. Rasayan J. Chem. 11, 844–849. doi:10.31788/rjc.2018.1122097
Yue,G.L.,Chan,B.C.L.,Hon,P.M.,Kennelly,E.J.,Yeung,S.K.,etal.(2010).
Immunostimulatory Activities of Polysaccharide Extract Isolated from Curcuma
longa.Int. J. Biol. Macrol. 47, 42–347. doi:10.1016/j.ijbiomac.2010.05.019
Yuliawati, T. H., and Hestinah, E. P. (2010). Cytotoxicity Effect of Curcuma
aeruginosa Extract on Fibroblast with MTT Assay Method. Folia Med.
Indonesiana. 46, 120–124.
Zhernakova, A., van Diemen, C. C., and Wijmenga, C. (2009). Detecting Shared
Pathogenesis from the Shared Genetics of Immune-Related Diseases. Nat. Rev.
Genet. 10, 43–55. doi:10.1038/nrg2489
Zhou, L., Zhang, K., Li, J., Cui, X., Wang, A., Huang, S., et al. (2013). Inhibition of
Vascular Endothelial Growth Factor-Mediated Angiogenesis Involved in
Reproductive Toxicity Induced by Sesquiterpenoids of Curcuma zedoaria in
Rats. Reprod. Toxicol. 37, 62–69. doi:10.1016/j.reprotox.2013.02.001
Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest.
Copyright © 2021 Yuandani, Jantan, Rohani and Sumantri. This is an open-access
article distributed under the terms of the Creative Commons Attribution License (CC
BY). The use, distribution or reproduction in other forums is permitted, provided the
original author(s) and the copyright owner(s) are credited and that the original
publication in this journal is cited, in accordance with accepted academic practice.
No use, distribution or reproduction is permitted which does not comply with
these terms.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311925
Yuandani et al. Immunomodulatory Effects of Curcuma Species

GLOSSARY
ADMET absorption, distribution, metabolism, elimination, and toxicity
ALP alkaline phosphatase
ALT alanine amino transferase
AP-1 activator protein-1
AMPs antimicrobial peptides
APC antigen-presenting cells
AST aspartate aminotransferase
C5aR complement C5a receptor
CL chemiluminescence
CMS chronic mild stress
CO
2
carbon dioxide
CRP C-reactive protein
DTH delayed hypersensitivity
EOCL essential oil of C. longa
fMLP formyl methionyl-leucyl-phenylalanine;
fMLPR formyl-methionyl-leucyl-phenylalanine receptor
GOT glutamic oxaloacetic transaminase
HFD- high-fat diet-
HOCl hypochlorous acid
HUVEC human umbilical vein endothelial cells
IBV infectious bronchitis virus
IBD inflammatory bowel diseases
IgM immunoglobulin M
IL-12 Interleukin 12
IL-6 interleukin-6
IL-1βinterleukin-1β
iNOS inducible nitric oxide synthase
ISI Institute for Scientific Information
IκB-αI kappa B alpha
LPS lipopolysaccharide
MAPKs mitogen-activated protein kinases
MCHC mean corpuscular hemoglobin concentration
MCH mean corpuscular hemoglobin
MCP1 monocyte chemoattractant protein-1
MCV mean corpuscular volume
MIP1αmacrophage inflammatory protein-1α
MHC major histocompatibility complex
MPO myeloperoxidase
NO nitric oxide
NF-ĸBnuclear factor-kappa B
PAFR platelet-activating factor receptor
PGE2 prostaglandin E2
PHA-P phytohemagglutinin-P
PMNs polymorphonuclear cells
PMA phorbol 12-myristate 13-acetate
PUFAs polyunsaturated fatty acids
ROS reactive oxygen species
SLE systemic lupus erythematosus
sRBCs sheep red blood cells
T1D type 1 diabetes
TLC total leukocytes number
TNF-αtumor necrosis factor-alpha.
Frontiers in Pharmacology | www.frontiersin.org April 2021 | Volume 12 | Article 64311926
Yuandani et al. Immunomodulatory Effects of Curcuma Species