![Efficacy of using keyhole limpet hemocyanin (KLH) and GNPs as nanocarriers. (a) Antibodies obtained by using KLH recognize the epitopes of both the antigen and the carrier protein. (b) Antibodies obtained by using GNPs recognize only the antigen's epitopes [80].](https://www.researchgate.net/profile/Lev-Dykman/publication/340785608/figure/fig1/AS:892428219334656@1589782876134/Efficacy-of-using-keyhole-limpet-hemocyanin-KLH-and-GNPs-as-nanocarriers-a_Q320.jpg)
![Scheme of vaccine design. M2e is conjugated to GNPs. By keeping M2e in excess in the solution, complete surface-coverage of GNPs with M2e is ensured at all times. Soluble CpG is added to the conjugate [86].](https://www.researchgate.net/profile/Lev-Dykman/publication/340785608/figure/fig2/AS:892428219338752@1589782876180/Scheme-of-vaccine-design-M2e-is-conjugated-to-GNPs-By-keeping-M2e-in-excess-in-the_Q320.jpg)
![Average antibody titers obtained with various immunostimulant combinations [94].](https://www.researchgate.net/profile/Lev-Dykman/publication/340785608/figure/fig3/AS:892428219318272@1589782876234/Average-antibody-titers-obtained-with-various-immunostimulant-combinations-94_Q320.jpg)
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Expert Review of Vaccines
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Gold nanoparticles for preparation of antibodies
and vaccines against infectious diseases
Lev A. Dykman
To cite this article: Lev A. Dykman (2020): Gold nanoparticles for preparation of
antibodies and vaccines against infectious diseases, Expert Review of Vaccines, DOI:
10.1080/14760584.2020.1758070
To link to this article: https://doi.org/10.1080/14760584.2020.1758070
Accepted author version posted online: 19
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REVIEW
Gold nanoparticles for preparation of antibodies and vaccines against infectious
diseases
Lev A. Dykman
Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov, Russia
ABSTRACT
Introduction: Vaccination remains very effective in stimulating protective immune responses against
infections. An important task in antibody and vaccine preparation is to choose an optimal carrier that
will ensure a high immune response. Particularly promising in this regard are nanoscale particle carriers.
An antigen that is adsorbed or encapsulated by nanoparticles can be used as an adjuvant to optimize
the immune response during vaccination. a very popular antigen carrier used for immunization and
vaccination is gold nanoparticles, with are being used to make new vaccines against viral, bacterial, and
parasitic infections.
Areas covered: This review summarizes what is currently known about the use of gold nanoparticles as
an antigen carrier and adjuvant to prepare antibodies in vivo and design vaccines against viral, bacterial,
and parasitic infections. The basic principles, recent advances, and current problems in the use of gold
nanoparticles are discussed.
Expert opinion: Gold nanoparticles can be used as adjuvants to increase the effectiveness of vaccines
by stimulating antigen-presenting cells and ensuring controlled antigen release. Studying the charac-
teristics of the immune response obtained from the use of gold nanoparticles as a carrier and an
adjuvant will permit the particles’potential for vaccine design to be increased.
ARTICLE HISTORY
Received 22 January 2020
Accepted 16 April 2020
KEYWORDS
Vaccine; gold nanoparticles;
nanocarriers; adjuvant;
immunization; antibodies;
infections
1. Introduction
Vaccination remains very effective in stimulating protective
immune responses against infections. Among the most
advanced diagnostic tools is the immunochemical method,
based on the use of highly pathogen-specific antibodies. An
important task in antibody and vaccine preparation is to
choose an optimal carrier (delivery system) that would ensure
a high immune response. Such adjuvant carriers can deposit
antigens at the injection sites, enhance their presentation to
immunocompetent cells, and induce the production of neces-
sary cytokines. Particularly promising in this regard are various
nanoscale particle carriers [1–16]. Such nanocarriers can be
used as adjuvants in the preparation of antibodies and next-
generation vaccines. The term ‘nanovaccinology’[17] is com-
ing into common scientific use.
Of note, the stimulation of antibody production in response
to antigens adsorbed on colloidal particles was discovered as
early as the beginning of the twentieth century [18–20]. In the
same period, studies began on the effect of colloidal metal
particles on the cells of the immune system [21,22]. In recent
decades, significant progress has been made in understanding
the immunogenicity of nanoparticles, the response of the
immune cells, the nature of immunomodulation and immuno-
suppression, and the cyto- and immunotoxicity of nanoparti-
cles, with account taken of their physical–chemical properties
[23–38].
Cells of the immune system constitute the first barrier to
the entry of nanoparticles into animal tissues and cells. In this
context, of undoubted interest is the study of the interaction
of nanoparticles with phagocytic cells, the mechanisms of
intracellular penetration, and the response of immune cells
to nanoparticles [39]. Depending on their size, nanoparticles
penetrate macrophages through receptor-mediated endocy-
tosis (phagocytosis) or pinocytosis and are localized mainly in
lysosomes and in perinuclear space. Often, they are wrapped
inside fringed vesicles. Some researchers believe that a key
part in nanoparticle uptake by macrophages is played by
scavenger receptors [40,41]. Nanoparticle penetration into
macrophages may enhance the respiratory activity of the
cells and, in some cases, the release of inflammatory mediators
(cytokines, prostaglandins, etc.) and the activation of the
immune response genes [42,43].
Of great interest are studies on the penetration of nano-
particles not only into macrophages but also into other
immune system cells, in particular dendritic cells. Dendritic
cells treated with gold nanoparticles can affect the activation
of CD8
+
T cells, which leads to epitope-specific cytotoxic
T-lymphocyte responses in vitro [44]. Compared with the use
of the native antigen, the internalization of antigen-
conjugated nanoparticles into dendritic cells increases the
immune response, enhancing lymphocyte proliferation [45].
In addition, nanoparticles can activate other immune system
CONTACT Lev A. Dykman dykman_l@ibppm.ru Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences,
Saratov, Russia
EXPERT REVIEW OF VACCINES
https://doi.org/10.1080/14760584.2020.1758070
© 2020 Informa UK Limited, trading as Taylor & Francis Group
cells –neutrophils [46,47], lymphocytes [48,49], and mono-
cytes [50,51].
Thus, nanoparticle penetration into the immune cells,
which causes cytokine induction, stimulation of T cells, activa-
tion of the immune response genes, enhanced antigen pro-
cessing, and antibody secretion by B cells, offers a good
possibility of using nanoparticles as carriers and adjuvants in
the preparation of antibodies and vaccines against infections
[52–55]. In particular, various nanoparticles are being used to
make new vaccines against viral [56–60], bacterial [61], and
parasitic [62] infections. On the basis of nanocarriers, subunit
[63] and peptide [64] vaccines are being developed for the
oral [65], intranasal [66], and percutaneous [67] routes of
administration.
A very popular antigen carrier used for immunization and
vaccination is gold nanoparticles (GNPs) [68–71]. Owing to
their unique physicochemical properties, ease of preparation,
and low toxicity, GNPs are widely used in various fields of
biomedical research [72]. For the first time, GNPs were used
as carriers of haptens for antibody preparation in 1986 [73].
Since then, a large number of reports have been published in
which this method was used and improved to develop anti-
bodies to a number of haptens and complete antigens [70]. It
was found that adjuvant properties are inherent in GNPs
themselves [74–76].
This review summarizes what is now known about the use
of GNPs to prepare antibodies and vaccines against viral,
bacterial, and parasitic infections.
2. Gold nanoparticles in the design of antiviral
vaccines
GNPs were first used to develop vaccines by Demenev et al. in
1996 [77], who examined the protective properties of an
experimental tick-borne encephalitis vaccine. For tick-borne
encephalitis virus and other flaviviruses, a slowly sedimenting
virus-specific antigen was described that had complement-
binding activity, was not associated with the virion fraction,
and did not have infectivity or hemagglutinating activity. This
antigen was called a soluble antigen [78]. The vaccine was
prepared by conjugating GNPs (average diameter, 15 nm) to
the soluble antigen. White laboratory mice were vaccinated
intraperitoneally three times, each at 32 μg of antigen per
injection. The protective properties of the experimental and
commercial vaccines were compared on the basis of the aver-
age survival time and the protection coefficients after vaccina-
tion of mice challenged with a virus-containing suspension
(100,000 and 10,000 LD
50
). The survival time and the thera-
peutic and protective efficacy obtained when the experimen-
tal vaccine was used were about 1.5 times greater than when
the commercial vaccine was used.
Many studies on GNP immunization and vaccination have
used such important objects as influenza virus and foot and
mouth disease (FMD) virus. Highly specific antibodies were
obtained from the immunization of animals with 17-nm
GNPs coupled to the pFMDV and pH5N1 antigens of these
viruses [79]. BALB/C white mice were vaccinated intraperito-
neally. Both antisera obtained had a high titer. The same
research team evaluated the effect of GNP size on the immune
response to the synthetic FMD peptide pFMDV [80]. GNPs with
diameters of 2, 5, 8, 12, 17, 37, and 50 nm were used. BALB/C
mice were immunized intraperitoneally six times, each at 1 μg
of antigen per injection. The antigens were mixed with com-
plete Freund’s adjuvant (CFA). The antibody titer was maximal
for 8-, 12- and 17-nm GNPs. The titer of the antisera obtained
by using GNPs was threefold higher than when keyhole limpet
hemocyanin (KLH) was used as an adjuvant and did not
require further purification from contaminant antibodies
(Figure 1). The amount of GNPs that had accumulated in the
spleen correlated with the magnitude of the immune
response elicited by the GNP/pFMDV conjugate.
Double intraperitoneal immunization of rats with 15-nm
GNPs complexed with the M1 antigen of the influenza virus
of the attenuated strain PR8 at a dose of 20 mg/kg produced
high-titer antibodies [81]. The production of IFN-γand inter-
leukins (ILs) 1βand 6 was stimulated, and the respiration of
peritoneal macrophages and spleen lymphocytes, was acti-
vated. Similar results were produced by double subcutaneous
immunization of guinea pigs with 12 μg of 15-nm GNPs con-
jugated to a synthetic VP
1
peptide of the FMD virus capsid
[82]. The titer of the resultant antibodies was not lower than
that obtained with CFA.
When guinea pigs were injected with FMD viruslike parti-
cles complexed with gold nanostars (50 μg, single injection),
the titer of specific and neutralizing antibodies and the stimu-
lation of T lymphocyte proliferation were much greater than
Article Highlights
●Gold nanoparticles have been used to prepare antibodies and vac-
cines against more than 45 pathogens of viral, bacterial, and parasitic
infections.
●The antibody elaboration during immunization with antigens conju-
gated to gold nanoparticles enhances the secretion of cytokines.
●Vaccination of animals with antigens conjugated to gold nanoparti-
cles decreases the symptoms of infections and protects the animals
challenged with virulent pathogens (up to lethal doses).
●By ‘programming’the activation state of dendritic or other antigen-
presenting cells, nanoparticles directly affect the induction of cellular
and humoral immunity.
●Studying the characteristics of the immune response obtained from
the use of gold nanoparticles as a carrier and an adjuvant will permit
the particles’potential for vaccine design to be increased.
Figure 1. Efficacy of using keyhole limpet hemocyanin (KLH) and GNPs as
nanocarriers. (a) Antibodies obtained by using KLH recognize the epitopes of
both the antigen and the carrier protein. (b) Antibodies obtained by using GNPs
recognize only the antigen’s epitopes [80].
2L. A. DYKMAN
those in guinea pigs immunized with the antigen in the pre-
sence of mineral oil ISA206 as an adjuvant. The proposed
nanovaccine more effectively protected virus-infected animals
[83].
A prototype of the intranasal influenza a vaccine was first
proposed by Gill’s group [84]. The synthetic peptide M2e,
a conserved extracellular domain of influenza virus matrix
protein 2 conjugated to 12-nm GNPs, was used as an immu-
nogen (Figure 2). As an adjuvant, soluble CpG (cytosine–phos-
phate–guanosine) oligodeoxynucleotide (CpG ODN) was used.
BALB/C mice were immunized twice at an interval of 21 days,
each time with 8.2 μg of M2e per animal. The conjugate
induced the production of specific IgG (with stimulation of
subtypes of both IgG1 and IgG2a), which could recognize M2e
and native M2 on influenza a viruses and had a protective
effect against a lethal dose of PR8-H1N1 virus. Subsequently,
Gill’s group published more detailed results from the use of
the proposed prototype vaccine to induce protective immu-
nity against the influenza a virus serotypes H1N1, H3N2, and
H5N1 [85,86]. Bimler et al. [87] showed that even at 15 months
after being vaccinated with the GNP/M2e+CpG conjugate, the
mice retained high titers of M2e-specific antibodies. These
mice were protected against lethal H1N1 influenza. In addi-
tion, the antibody titers increased after a challenge with influ-
enza a and remained elevated for 3 months. This suggests that
old mice retain effective M2e-specific memory B cells.
When mice were immunized with the M2e antigen of influenza
a virus, the most effective complex immunogens were M2e +
GNPs and CpG + GNPs [88]. When they were used, the titer of
the obtained antibodies was the highest. In addition, the immu-
nogens activated the respiratory activity of lymphoid cells and the
production of proinflammatory cytokines to a greater extent than
did other immunogens, including a commercial vaccine. The adju-
vant effect of GNPs is probably associated with the more efficient
penetration of the conjugate into phagocytic cells, which leads to
improved presentation of the antigen to antibody-forming cells.
In general, GNPs as an antigen carrier and adjuvant are
often combined with other immune response stimulants,
most commonly with CFA and CpG ODNs. It is known that
when CpG motifs are unmethylated, they act as immunosti-
mulants [89]. The immunostimulatory activity of synthetic
ODNs containing CpG sequences may be similar to the activity
of bacterial DNA [90]. According to Wei et al. [91], GNPs
coupled to CpG ODNs effectively enhance the intracellular
penetration of nanoparticles into macrophages and signifi-
cantly increase the secretion of proinflammatory cytokines –
tumor necrosis factor α(TNF-α) and IL-6. Moreover, the
immunostimulatory effect of GNPs conjugated to CpG ODNs
is often significantly higher than that of the same concentra-
tions of soluble CpG ODNs [92,93]. Of note, the optimal immu-
nization method is the simultaneous use of two conjugates:
GNPs–antigen and CpG ODNs–GNPs [94](Figure 3). The
immune response to an intranasal influenza vaccination can
be enhanced with gold nanorods coupled to polyinosinic-
polycytidylic acid as an adjuvant, as well as with CpG [95].
Wang et al. [96,97] proposed the use of two H3N2 influenza
aantigens for vaccination –hemagglutinin and flagellin –in
complex with 18-nm GNPs. In a single intranasal immuniza-
tion, both GNP/hemagglutinin/flagellin and GNP/hemaggluti-
nin complexes induced higher levels of antibody production
in mice than did a mixture of soluble hemagglutinin and
flagellin. Immunization elicited a strong mucosal and systemic
immune response, which protected the hosts from lethal
influenza. In addition, it was shown in vitro that the conjugates
promote the maturation of dendritic cells for the processing
and presentation of antigens, stimulate the production of
cytokines, and promote antigen-specific T-cell immunity.
GNPs were used to prepare antibodies to the surface anti-
gens of porcine transmissible gastroenteritis virus [98,99]. Mice
were immunized once intraperitoneally with 70 μg of antigen–
15-nm GNPs; guinea pigs, twice subcutaneously with 125 μg;
and rabbits, three times subcutaneously with 220 μg. The
resulting virus-neutralizing antibodies from all animal species
Figure 2. Scheme of vaccine design. M2e is conjugated to GNPs. By keeping M2e in excess in the solution, complete surface-coverage of GNPs with M2e is ensured
at all times. Soluble CpG is added to the conjugate [86].
Figure 3. Average antibody titers obtained with various immunostimulant
combinations [94].
EXPERT REVIEW OF VACCINES 3
had a much higher titer than did antibodies developed against
the native virus. Immunization with the GNP/antigen complex
significantly enhanced the respiratory activity of peritoneal
macrophages and spleen lymphocytes and increased the con-
tent of IFN-γ, IL-1β, and IL-6 in the blood plasma of the
immunized animals. In addition, there was an increase in the
number of follicles in the spleen, indicating the activation of
humoral immunity.
A conjugate of 15-nm GNPs with a glycoprotein isolated
from fixed rabies virus, strain Moscow 3253, was used to
develop highly specific antibodies to the virus [100]. In the
first immunization, 25 μg of the antigen was intraperitoneally
injected into mice; in the subsequent four immunizations,
50 μg was used. The immunogenicity of the purified glyco-
protein was higher than that of whole rabies virus, and the
glycoprotein conjugated to GNPs was best able to induce
virus-neutralizing antibodies. The resulting antibodies were
used to make a diagnostic agent based on solid-phase immu-
noassay. Antirabies virus antibodies were also prepared by
using the GNP-conjugated ribonucleoprotein of an attenuated
rabies virus [101].
To prepare antibodies against West Nile fever virus, Niikura
et al. [102] used antigen conjugates with GNPs of various sizes
and shapes: 20- and 40-nm nanospheres, 40 × 20-nm nanorods,
and 40 × 40 × 40-nm nanocubes. Mice were immunized twice
intraperitoneally with 100 ng of the conjugate per animal.
Forty-nm nanospheres induced the highest concentration of
specific antibodies, while the titer of the antibodies obtained
with the other GNPs was nearly 2 times lower. The macro-
phages and dendritic cells absorbed larger numbers of nanor-
ods, which suggests that the production of antibodies does not
depend on the efficiency of absorption of various GNPs.
Nanorods enhanced the production of IL-1βand IL-18, while
nanospheres and nanocubes enhanced that of TNF-α,IL-6,IL-
12, and granulocyte–macrophage colony-stimulating factor.
Stone et al. [103] examined the immune response to nanor-
ods coupled to the glycoprotein antigen of respiratory syncy-
tial virus. The conjugate-treated human dendritic cells induced
an immune response (proliferation) of primary T cells.
The influence of the particle shape and size on the effec-
tiveness of the immune response was also investigated in [94].
Mice were injected with an antigen coupled to gold nano-
spheres (diameters, 15 and 50 nm), nanorods (70 × 15 nm),
nanoshells (SiO
2
core, 140 nm; gold shell, 27 nm), and nanos-
tars (core, 12 nm; rays, 15–25 nm). The antibody titers
increased from 1:160 to 1:10240 in the sequence native anti-
gen < nanorods < nanoshells < GNPs-15 nm < GNPs-50 nm. It
was concluded that spherical GNPs with diameters of 50 and
15 nm are the optimal antigen carrier and adjuvant for
immunization.
The V3βpeptide of the HIV-1 gp120 protein in complex with
2-nm gold glyconanoparticles was used as an immunogen to
prepareaprototypeHIVvaccine[104]. Rabbits were immunized
three times intramuscularly with 50 μgofthecomplex.The
resulting antibodies had a high titer and neutralizing activity
against HIV-1. The HIV-1 Gag p17 peptide conjugated to 2-nm
gold glyconanoparticles increased the proliferation of HIV-
specific CD4
+
and CD8
+
T cells and induced the secretion of
the highly functional TNF-αand IL-1βcytokines, as compared
to the unconjugated peptide [105]. Also, GNPs conjugated with
the gp120 and gp41 viral proteins were used as a platform for
thedeliveryofimmunogensinthepreparationofanHIV-1
vaccine [106].
Ten-nm GNPs were complexed simultaneously with CpG
ODN and with the recombinant hepatitis B virus core antigen
in the form of viruslike particles [107]. Mice were immunized
four times intraperitoneally with 50 μg of the conjugate. The
titer of the resultant antibodies was twofold higher than that
of the antibodies prepared without GNPs. The proliferation of
the CD4
+
and CD8
+
T cells, the secretion of IL-4 and IFN-γ, and
the stimulation of both Th1 and Th2 immune responses were
increased. That is, the conjugates showed a strong cellular and
humoral immunostimulatory ability.
The surface antigen of hepatitis B virus (HBsAg) was con-
jugated to gold nanocages [108]. The immunogenic properties
of the conjugate were examined in vitro in RAW 264.7 macro-
phages. The conjugate was intensely absorbed by the macro-
phages and contributed to the processing of the antigens and
to the secretion of IL-4.
For a prototype hepatitis E vaccine, vaccine monomers
were conjugated to gold fluorescent nanoclusters [109]. The
intraperitoneally injected conjugate not only enhanced the
Th1/Th2 immune response in mice but also reduced the toxi-
city of the vaccine. In addition, the own fluorescence of the
gold clusters made it possible to track the dynamic behavior
of the vaccine in vivo. GNPs were also used to prepare anti-
peptide antibodies to the binding sites of the E2 protein of
hepatitis C virus [110].
A prototype dengue virus vaccine was developed by using
gold nanorods functionalized with protein E virus (DENVE)
[111]. Immunization of mice with GNP/DENVE significant
increased IgG synthesis and splenocyte proliferation.
Quach et al. [112] conjugated differently sized GNPs to
domain III of envelope glycoprotein derived from serotype 2
of dengue virus (EDIII) as a dengue subunit vaccine. They used
GNPs with average particle diameters of 20, 40, and 80 nm.
The GNP/EDIII conjugate was used to immunize BALB/C mice
three times subcutaneously. The conjugate induced high con-
centrations of antibodies that provide serotype-specific neu-
tralization of dengue virus. The concentration of the
antibodies depended on both size and concentration of the
GNPs, and this made it possible to modulate the immune
response by adjusting these parameters. In addition, the con-
jugate activated the complement system, induced proliferative
responses in splenocytes and CD4
+
and CD8
+
T cells, and
induced the secretion of IL-4 and IFN-γ. Eighty-nm GNPs per-
formed the best.
Using the surface spike glycoprotein of avian coronavirus,
which causes infectious bronchitis in birds, Chen et al. [113]
obtained viruslike particles by incubating 100-nm GNPs in
a solution containing an optimized concentration of the viral
proteins. After free proteins were removed, the particles with
the antigen were shown by TEM to be morphologically similar
to the natural viral particles. BALB/C mice and specific-
pathogen-free chickens were vaccinated intramuscularly with
a single dose of 10 μg. Compared to free protein
4L. A. DYKMAN
immunization, conjugate vaccination showed improved anti-
gen delivery to lymphoid organs, higher antibody titers, an
increased response of spleen T cells, and a reduction in symp-
toms of coronavirus infection. a comparison with
a commercial whole inactivated viral vaccine also showed
that the GNP conjugate provided better antiviral protection.
GNPs conjugated to the S protein of severe acute respiratory
syndrome-related coronavirus induced a strong IgG response
upon immunization of mice [114].
GNPs were also used to make a conjugate with the antigen
of the Newcastle disease virus of birds [115]. Ross chickens
were vaccinated once subcutaneously with a mixture of GNPs
and the vaccine antigen. High-titer specific antibodies were
obtained, and increased secretion of IFN-γwas noted.
The capsid (Cap) protein of a pathogenic porcine circovirus
was conjugated to 23-nm GNPs [116]. In vitro studies showed
that GNPs contributed to Cap protein phagocytosis. Mice
immunized twice subcutaneously with GNP/Cap showed
high production of virus-neutralizing antibodies. Similar
results were obtained with classical swine fever virus anti-
gen [117].
Soliman et al. [118] investigated the use of gold nanospheres
and nanorods as an adjuvant to develop antibodies against the
Rift Valley fever virus. They noted that the immune response
obtained by a single subcutaneous immunization of rats
depended on the shape of the GNPs. Peak antibody concentra-
tions were detected on day 30 after immunization with nanorods
and on day 45 with nanospheres. However, the content of IFN-γ
and IL-10 was higher with nanospheres than it was with nanorods.
Research is also underway to design GNP-based vaccines
against other dangerous pathogens, such as Ebola, Zika,
Marburg, Lassa, and Nipah.
Table 1 summarizes the literature data on the viral antigens
that have been conjugated with GNP carriers and then used
for immunization of animals.
3. Gold nanoparticles in the preparation of
antibacterial vaccines
GNPs have also been intensely used to develop antibodies and
vaccines against bacterial infections. For the first time, 15-nm
GNPs were used as an adjuvant to prepare antibodies to the
surface antigens of Yersinia pseudotuberculosis [119]. Rabbits
received two subcutaneous injections, each with 1 μg of the
antigen. The serum titer was as high as when CFA was used,
but the amount of antigen injected when GNPs were used as
an adjuvant was smaller by two orders of magnitude. GNPs as
an antigen carrier activated the phagocytic activity of the
lymphoid cells. Similar results were obtained by the same
research team in the preparation of antibodies against
Y. enterocolitica and Brucella abortus.
The efficacy of 15-nm GNPs coated with Y. pestis F1 antigen
in animal immunization was evaluated by Gregory et al. [120].
The conjugate (0.93 μg) was administered to mice once
Table 1. Conjugates of GNPs with viral antigens used for immunization and vaccination of animals.
Viral antigen GNP size and shape
Functionalization approach
(ligand)
Chemical nature of
antigen Refs.
Tick-borne encephalitis virus Nanospheres, 15 nm Adsorption Isolated protein [77]
Foot and mouth disease virus Nanospheres, 17 nm Chemosorption (cysteine) Synthetic peptide [79]
-
”
- Nanospheres, 2, 5, 8, 12, 17, 37, and 50 nm Chemosorption (cysteine) Synthetic peptide [80]
-
”
- Nanospheres, 15 nm Adsorption Synthetic peptide [82]
-
”
- Nanostars Adsorption Virus-like particles [83]
Influenza virus Nanospheres, 17 nm Chemosorption (cysteine) Synthetic peptide [79]
-
”
- Nanospheres, 15 nm Adsorption Isolated peptide [81]
-
”
- Nanospheres, 12 nm Adsorption Synthetic peptide [84]
-
”
- Nanospheres, 12 nm Chemosorption (cysteine) Synthetic peptide [87]
-
”
- Nanospheres, 15 nm Adsorption Synthetic peptide [88]
-
”
- Nanospheres, 18 nm Chemosorption (N
3
-PEG-SH,
SH-NTA)
Recombinant
proteins
[96]
Porcine transmissible gastroenteritis virus Nanospheres, 15 nm Adsorption Isolated protein [99]
Rabies virus Nanospheres, 15 nm Adsorption Isolated glycoprotein [100]
-
”
- Nanospheres, 15 nm Adsorption Isolated
ribonucleoprotein
[101]
West Nile fever virus Nanospheres, 20 and 40 nm; Nanorods, 40 × 20 nm;
Nanocubes, 40 nm
Chemosorption (PSS-MA) Isolated protein [102]
Respiratory syncytial virus Nanorods, 21 × 57 nm Chemosorption (EDC) Recombinant
glycoprotein
[103]
HIV Glyconanoparticles, 2 nm Adsorption Synthetic peptide [104]
Hepatitis B virus Nanospheres, 10 nm Adsorption Recombinant protein [107]
-
”
- Nanocages, 50 nm Adsorption Recombinant protein [108]
Hepatitis E virus Nanoclusters Direct synthesis Hepatitis E vaccine [109]
Hepatitis C virus Nanospheres, 15 nm Adsorption Synthetic peptide [110]
Dengue virus Nanospheres, 20, 40, and 80 nm Adsorption Recombinant
glycoprotein
[112]
Avian coronavirus Nanospheres, 100 nm Adsorption Recombinant protein [113]
Severe acute respiratory syndrome-related
coronavirus
Nanospheres, 40 nm Chemosorption (BSPP) Recombinant protein [114]
Newcastle disease virus Nanospheres Mixing Newcastle disease
vaccine
[115]
Porcine circovirus Nanospheres, 23 nm Adsorption Recombinant protein [116]
Classical swine fever virus Nanospheres, 24 nm Adsorption Recombinant protein [117]
Rift Valley fever virus Nanospheres, 20 nm; Nanorods, 40 × 20 nm Adsorption Isolated protein [118]
EXPERT REVIEW OF VACCINES 5
intramuscularly. The animals injected with GNP/F1 generated
the greatest IgG antibody response to the F1 antigen, as
compared to those injected with unconjugated F1 antigen in
phosphate-buffered saline or in Alhydrogel®. Sera obtained
against the F1 antigen coupled to GNPs were able to bind to
the F1 antigen in an enzyme-linked immunoassay. Similar
results were produced by Kireev et al. [121].
GNPs decorated with thiolated glycosides (synthetic analogs of
the capsular polysaccharides of Neisseria meningitidis)werepro-
posed as a synthetic antigen for use in immunization [122].
Conjugates of 15-nm GNPs with Salmonella typhimurium surface
antigens stimulated macrophage respiratory activity and the activ-
ity of the mitochondrial enzymes succinate dehydrogenase and
glycerophosphate dehydrogenase [42]. This stimulation may be
a significant factor determining the adjuvant properties of GNPs.
GNPs were included in the transmucosal delivery of tetanus
toxoid, used for vaccination against tetanus, a deadly disease
caused by Clostridium tetani [123]. Subcutaneous injection of
tetanus toxoid conjugated to 25-nm GNPs into mice gener-
ated a systemic response but did not cause any mucosal
response. However, the conjugate generated a significantly
higher mucosal response after oral administration. For enhan-
cing the immune response to tetanus toxoid, plant adjuvants
(saponins) from Quillaja saponaria [124] and Asparagus race-
mosus [125] were used together with GNPs. GNPs were also
used in the preparation of a vaccine against botulism,
a particularly dangerous disease caused by C. botulinum [126].
GNPs (average diameter, ~ 2 nm) were conjugated to a syn-
thetic tetrasaccharide epitope, an analog of the capsular polysac-
charide of Streptococcus pneumoniae type 14 [127,128]. The
obtained glyconanoparticles conjugated with T-helper peptide
generated specific high-titer IgG after a single intradermal immu-
nization of mice with 3 μg of the conjugate. An increase in the
content of the cytokines IL-2, IL-4, IL-5, IL-17, and IFN-γconfirmed
that the nanoparticles activate T helpers. The antisaccharide anti-
bodies stimulated the phagocytosis of type 14 bacteria by human
leukocytes, which indicates the functionality of the antibodies.
A new strategy for preparing antibodies to tuberculin was
described [129] that used the adjuvant properties of GNPs.
Tuberculin is a mixture of the surface antigens of various types
of mycobacteria (Mycobacterium tuberculosis, M. bovis,and
M. avium). When injected intradermally, it causes a specific
delayed-type hypersensitivity response in infected or vaccinated
patients. Polyclonal antituberculin antibodies were raised by
injecting 7.5 μg of tuberculin conjugated to 15-nm GNPs into
rabbits four times intramuscularly. The obtained antibodies had
a high titer, whereas in the control animals, which were given
nonconjugated tuberculin, no antibodies were found in the
blood serum. The antituberculin antibodies were used to detect
mycobacteria by ELISA and by light and electron microscopy.
GNPs were found to partially remove the toxic effect of
tuberculin on rat peritoneal macrophages owing to their
penetration into the intracellular space. This decrease in toxi-
city contributed to the better development of the humoral
response and to the elaboration of antituberculin antibodies.
In addition, GNPs decorated with tuberculin enhanced the
adhesion of phagocytic cells to microbial cells. Preliminary
vaccination results for GNPs conjugated to guinea pig tuber-
culin showed a protective effect [129].
For immunoassay of mycobacteria, GNPs were conjugated
to the penta- and hexasaccharide fragments of mycobacterial
lipoarabinomannan by using aglycon spacers of various
lengths [130]. Rabbit polyclonal antibodies to these conju-
gates were developed. The antisera obtained by hyperimmu-
nization of rabbits detected with a high titer both
lipoarabinomannan oligosaccharides and mycobacterial cells.
Gao et al. [131] coated 30-nm GNPs with bacterial vesicles of
the outer membrane of Escherichia coli (Figure 4). When injected
three times subcutaneously, the conjugate induced rapid activa-
tion and maturation of dendritic cells in the lymph nodes of the
vaccinated mice. In addition, vaccination with the conjugate
gave rise to antibodies that had a higher avidity than those
obtained by vaccination with outer membrane vesicles only.
The production of IFN-γand IL-17 increased too, which indicates
strong Th1 and Th17 cellular responses to the bacteria.
Another prototype vaccine against E. coli was demon-
strated by Sanchez-Villamil et al. [132]. GNPs were coated
with the specific immunogenic LomW and EscC antigens of
the enterohemorrhagic strain E. coli O157: H7. Mice were
injected three times subcutaneously with GNPs/LomW, GNPs/
EscC, or a combination thereof, containing equivalent
amounts of both immunogens. As a result, higher-titer IgG
were found in sera and higher-titer IgA were found in feces.
Serum IgG titers correlated with a significant decrease in the
intestinal colonization of E. coli at 3 days after inoculation. The
sera from the mice immunized with antigen-coated GNPs
reduced the adhesion of E. coli O157: H7 and two other
E. coli pathotypes to human intestinal epithelial cells. In addi-
tion, the sera had antigen-specific bactericidal properties.
An effective vaccine against the human pathogen Listeria
monocytogenes was made by conjugating the listeriolysin
O peptide LLO91-99 to gold glyconanoparticles [133].
a single intraperitoneal or intravenous immunization of mice
with the conjugate induced a specific T-cell response and
protected the animals against Listeria infection. The protection
correlated with an increase in the number of splenic CD4
+
and
CD8
+
T cells, NK cells, and CD8α
+
dendritic cells, and it also
correlated with an increase in the production of the cytokines
IL-12, IFN-γ, TNF-α, and MCP-1 after infection. Subsequently, it
was shown that vaccination of pregnant mice with gold gly-
conanoparticles carrying L. monocytogenes peptides protects
Figure 4. Schematic illustration of modulation of antibacterial immunity with
bacterial-membrane-coated nanoparticles [130].
6L. A. DYKMAN
newborns from listeriosis [134]. Newborn mice born to vacci-
nated females were free of bacteria and healthy, whereas
nonvaccinated mice had obvious lesions of the brain and skin.
An interesting vaccination schedule against glanders,
a disease caused by Burkholderia mallei, was proposed by
Gregory et al. [135]. GNPs (average diameter, 15 nm) were
first covalently bound to recombinant protein carriers –the
Hc fragment of tetanus toxin, the hemolysin coregulated pro-
tein produced by both B. mallei and B. pseudomallei, and the
flagellin produced by B. pseudomallei.Theconjugatessopre-
pared were functionalized with purified LPS from a nonvirulent
B. thailandensis strain. Mice were immunized three times intra-
nasally, and the dose used was 0.93 μg. The GNP/protein/LPS
conjugates generated significantly higher antibody titers than
did native LPS. In addition, they improved protection against
a lethal inhalation challenge of B. mallei in the murine model of
infection. The proposed scheme in the form of aerosol immu-
nization was successfully tested on rhesus monkeys [136].
a similar approach was developed for vaccination against
B. pseudomallei myeloidosis [137]. Mice immunized three
times subcutaneously with GNPs/protein/LPS generated high-
titer antibodies. Importantly, the immunized animals survived
nearly 100% and their lungs were less contaminated with bac-
teria after a lethal infection with B. pseudomallei.
Dakterzada et al. [138] evaluated the efficacy of GNPs con-
jugated with the N-terminal domains of Pseudomonas aeruginosa
flagellin as an immunogen in the vaccination of mice.
Nanoparticles were complexed with a recombinant flagellin (1–-
161). Flagellin, GNP/flagellin, and flagellin emulsified with CFA
were administered twice subcutaneously to BALB/C mice at
adoseof10μg. The mice given GNPs/flagellin elaborated
more antiflagellin antibodies than did those that either were
not immunized at all or were given adjuvant-free flagellin. In
a whole-cell ELISA, these antibodies efficiently recognized native
flagellin on the bacterial cells. An assay of opsonophagocytosis
demonstrated the functional activity and specificity of the anti-
flagellin antibodies prepared by using the GNP/flagellin conju-
gate against the homologous strain.
The acidic serine protease AprV2 secreted by virulent
Dichelobacter nodosus, the causative agent of infectious podo-
dermatitis, was conjugated to 20-nm GNPs [139]. The nano-
vaccine induced higher IgG responses than did native AprV2
or the vaccine adjuvanted with monophosphoryl lipid A.
Conjugates of 15-nm GNPs with two isolated Francisella
tularensis antigens, a protective antigenic complex and
a glycosylated protein complex, were used to obtain antitular-
emia sera and vaccinate animals [140]. Subcutaneous immu-
nization of mice with GNPs decorated with a glycosylated
protein complex was more effective than was immunization
with an unconjugated antigen. This greater effectiveness was
manifested as an increase in the protection ability and titers of
antibodies. The conjugation of GNPs to both antigens in the
immunization of rabbits afforded sera with a high titer of
specific antibodies in a relatively short period and with mini-
mal consumption of the antigen. The use in ELISA of the
immunoglobulins isolated from the sera allowed detection of
F. tularensis cells of different subspecies, enabling their further
use in the manufacture of diagnostic agents for tularemia.
Recombinant immunodominant antigens of Coxiella burne-
tii, the causative agent of Q fever, were conjugated to GNPs
[141]. Immunization of mice and guinea pigs gave rise to
specific antibodies and activated the response of the CD4
+
T cells. The protective effect of the GNP-associated antigens
was comparable to that of the commercial vaccine used.
Conjugates of 15-nm GNPs with seven Vibrio cholerae anti-
gens were synthesized in [142]. Rabbits were immunized five
times subcutaneously, and the resultant polyclonal antibodies
to the test antigens showed a high specific activity. By using
white laboratory mice, the protective activity of the conju-
gates was evaluated during infection of vaccinated animals,
with a commercial vaccine as a control. The cholera vaccine
prototypes made with GNPs as a carrier and an adjuvant
corresponded to the commercial chemical vaccine in terms
of protection efficiency.
Table 2 summarizes the literature data on the bacterial
antigens that have been conjugated with GNP carriers and
then used for immunization of animals.
4. Gold nanoparticles in the preparation of
antiparasitic vaccines
Besides being used as an adjuvant in immunization against
viral and bacterial diseases, GNPs are also used to develop
antibodies and vaccines against parasitic infections. Parween
et al. [143] reported that 17-nm GNPs in complex with recom-
binant peptides of the C-terminal 19kDa fragment of mero-
zoite surface protein 1 from the malaria pathogen Plasmodium
falciparum had a pronounced immunogenic activity when
combined with the adjuvant Alhydrogel®. Mice were immu-
nized three times subcutaneously at a dose of 25 μg.
Antibodies against the weakly immunogenic peptides conju-
gated to GNPs recognized the native protein on the parasite’s
surface and inhibited the invasion of P. falciparum in an in vitro
assay.
Another immunogen was proposed to make an antimalarial
vaccine –the P. falciparum surface protein Pfs25, expressed in
zygotes and ookinetes [144]. The recombinant protein Pfs25
was attached to various GNPs, including nanospheres, nanos-
tars, nanocages, and nanoprisms. Mice were immunized with
the resulting conjugates three times intramuscularly at a dose
of 10 μg. This yielded high-titer antibodies, the highest titers
being obtained with gold nanospheres and nanostars. In
membrane feeding assays, the antibodies blocked the trans-
mission of parasites to mosquitoes.
GNPs were also used to prepare anti-idiotypic antibodies [145].
These antibodies were used for an indirect and a competitive
ELISA of the opisthorchiasis pathogen Opisthorchis felineus.Work
is underway to develop a vaccine against Babesia canis,thecau-
sative agent of canine piroplasmosis, by using the adjuvant prop-
erties of GNPs.
For vaccination against schistosomiasis, a parasitic disease
caused by Schistosoma mansoni trematodes, gold nanorods
were complexed with the recombinant protein rSm2, located
on the surface of larvae and adult worms [146]. Mice were
immunized three times intraperitoneally with 2 μg of the
conjugate. After immunization, a Th1 immunological response
EXPERT REVIEW OF VACCINES 7
was observed with higher production of IFN-γ, mainly by CD4
+
and CD8
+
T cells. In addition, the conjugate activated dendritic
cells in vitro, enhancing the expression of MHCII and MHCI and
the production of IL-1β. After vaccination with the conjugate,
followed by a challenge with S. mansoni cercariae, the number
of worms in the liver’s portal vein and the number of eggs in
the liver tissues decreased, as compared with the control
group.
Table 3 summarizes the literature data on the parasite
antigens that have been conjugated with GNP carriers and
then used for immunization of animals.
5. Expert opinion
GNPs have been used to prepare antibodies and vaccines
against more than 45 pathogens of viral, bacterial, and para-
sitic infections. The sizes of the GNPs used for this purpose
have ranged from 2 to 100 nm. Moreover, in most studies,
particle diameters ranging from 15 to 50 were recognized as
optimal. Variously shaped GNPs have been used, including
nanospheres, nanorods, nanocages, nanostars, nanocubes,
nanoshells, nanoprisms, and nanoclusters. Most researchers
have indicated that the best effect is achieved with gold
nanospheres as antigen carriers.
Various methods have been used to functionalize GNPs with
target antigens. Conjugation has been carried out both by
physical adsorption and by chemisorption. Passive adsorption
of an antigen on the particle surface occurs because of the
electrostatic and hydrophobic interactions. Coulomb
interactions between the H
2
N groups of lysine and citrate
ions on the surface of gold nanoparticles have also been
reported. Recently, information has appeared about an impor-
tant role of the SH groups of cysteine molecules in protein
binding to the surface of gold particles. It is known that
between sulfur and gold atoms, there arises a donor–acceptor
(semipolar) bond (dative bound). This fact has led to the use of
alkanethiol linkers (chemisorption method) for more stable
attachment of biomolecules to GNPs. In addition, GNPs have
often been conjugated to protein or polysaccharide linkers
before being functionalized with an antigen. Most often, gold
glyconanoparticles have been used as immunomodulators.
What other adjuvants can be used with GNPs remains an
open question. Specific antibodies against target antigens
have been obtained by using GNPs as the sole adjuvant in
combination with immune response enhancers such as CFA,
CpG-ODNs, MontanideTM, AdvaxTM, Alhydrogel®, Imiquimod,
monophosphoryl lipid A, ISA206, and saponins. Moreover,
both natural (native) antigens and genetically engineered
(recombinant) ones have been used as immunogens.
Various animals have been used for immunization and vac-
cination, including mice, rats, guinea pigs, rabbits, chickens, and
monkeys. The methods used to administer immunogens have
included intraperitoneal, subcutaneous, intradermal, intramus-
cular, intravenous, intranasal, and oral. Moreover, the immuni-
zation doses of the antigens varied within a very wide range –
for example, from 100 ng to 50 μg for mice or from 1 μgto
220 μg for rabbits. The frequency of administration also varied –
from single to sixfold.
Table 2. Conjugates of GNPs with bacterial antigens used for immunization and vaccination of animals.
Bacterial antigen GNP size and shape Functionalization approach (ligand) Chemical nature of antigen Refs.
Yersinia pseudotuberculosis Nanospheres, 15 nm Adsorption Isolated protein [119]
Yersinia pestis Nanospheres, 15 nm Chemosorption (NHS, EDC) Recombinant protein [120]
-
”
- Nanospheres, 15 nm Adsorption Isolated protein [121]
Neisseria meningitides*Nanospheres, 5 nm Chemosorption (thiols) Synthetic carbohydrate [122]
Salmonella typhimurium Nanospheres, 15 nm Adsorption Isolated protein [42]
Clostridium tetani*Glyconanoparticles, 25 nm Adsorption Isolated protein [123]
Streptococcus pneumoniae*Glyconanoparticles, 2 nm Direct synthesis Synthetic carbohydrate [127]
Mycobacterium tuberculosis Nanospheres, 15 nm Adsorption Isolated peptide [129]
-
”
-* Nanospheres, 15 nm Adsorption Synthetic carbohydrate [130]
Escherichia coli Nanospheres, 30 nm Adsorption Isolated bacterial membrane [131]
-
”
- Nanospheres, 15 nm Chemosorption (MHDA) Recombinant proteins [132]
Listeria monocytogenes*Glyconanoparticles, 1.5 nm Direct synthesis Synthetic peptides [133]
Burkholderia mallei*Nanospheres, 15 nm Chemosorption (NHS, EDC) Isolated LPS [135]
Burkholderia pseudomallei Nanospheres, 15 nm Chemosorption (NHS, EDC) Isolated LPS [137]
Pseudomonas aeruginosa Nanospheres, 15 nm Adsorption Recombinant protein [138]
Dichelobacter nodosus Nanospheres, 20 nm Adsorption Recombinant protein [139]
Francisella tularensis Nanospheres, 15 nm Adsorption Isolated proteins [140]
Coxiella burnetii Nanospheres, 15 nm Adsorption Recombinant proteins [141]
Vibrio cholerae Nanospheres, 15 nm Adsorption Isolated proteins and LPS [142]
*
Glyconanovaccine
Table 3. Conjugates of GNPs with parasite antigens used for immunization and vaccination of animals.
Parasite antigen GNP size and shape
Functionalization approach
(ligand)
Chemical nature of
antigen Refs.
Plasmodium
falciparum
Nanospheres, 17 nm Adsorption Recombinant peptides [143]
-
”
- Nanospheres, 30 nm; nanostars, 50 nm; nanocages, 60 nm;
nanoprisms, 40 nm
Chemosorption (4-ATP) Recombinant protein [144]
Babesia canis Nanospheres, 15 nm Adsorption Isolated proteins [unpublished
data]
Schistosoma
mansoni
Nanorods Adsorption Recombinant protein [146]
8L. A. DYKMAN
The antibody elaboration during immunization with GNP–
antigen conjugates enhanced the secretion of cytokines –
most often, IFN-γ, TNF-α, IL-1β, IL-4, IL-6, IL-10, and IL-12,
and less often, IL-2, IL-5, IL-17, IL-18, MCP-1, and GM-CSF.
Several researchers have noted the stimulation of the respira-
tory activity of peritoneal macrophages and spleen lympho-
cytes and the activation of the macrophage mitochondrial
enzymes succinate dehydrogenase and glycerophosphate
dehydrogenase. Immunization with GNP–antigen conjugates
enhanced the proliferation of the CD4
+
and CD8
+
T cells and
NK cells and stimulated both Th1 and Th2 immune responses.
Other effects noted were the activation of dendritic cell
maturation for antigen processing and presentation, increased
expression of MHCII and MHCI, and activation of NLRP3
inflammasomes. The active uptake of GNPs by macrophages
contributed to the processing of antigens. Splenocytes of the
animals immunized with GNPs had higher proliferation rates
than the cells immunized with the native antigen alone, which
shows the ability of GNP-based carriers to induce enhanced
cellular responses to the associated antigens. Often, GNPs
decorated with bacterial antigens enhanced the phagocytic
adhesion of microbial cells.
Antibodies obtained by using GNPs have been successfully
applied to the immunochemical identification of pathogens
with various solid-phase and microscopic methods and with
biosensor devices. Vaccination of animals with GNP–antigen
conjugates decreased the symptoms of infections and pro-
tected the animals challenged with virulent pathogens (up
to lethal doses). In some cases, the vaccinated animals sur-
vived 100%. In most studies, the low toxicity of the gold carrier
itself was noted [147].
Currently, the immunostimulatory activity of nanomaterials,
including GNPs, as adjuvants is explained by the following
mechanisms (Figure 5). Antigen–nanocarrier conjugates are
more effectively delivered to the lymph nodes and induce
a depot effect, which contributes to the steady and gradual
release of the antigens. The use of nanoparticles as adjuvants
helps antigens to concentrate on the membrane of dendritic
cells and activates their maturation and the expression of
MHCI and MHCII. The spatial organization (multivalence) of
antigens on the surface of the particles enhances the prolif-
eration of T cells and the activation of B cells. Both events are
accompanied by the release of soluble mediators such as
cytokines, chemokines, and immunomodulating molecules,
which regulate the immune response.
Antigen-presenting cells are crucial for the response to
vaccines. Among the antigen-presenting cells, dendritic cells
are especially important for the primary immune response,
because they control the activation of the CD4
+
and CD8
+
T cells, which assist in the induction of antibodies and exhibit
direct cytotoxic activity. Therefore, the key application of
nanoparticles in immunology is the modulation of the func-
tions of antigen-presenting cells. By ‘programming’the activa-
tion state of dendritic or other antigen-presenting cells,
nanoparticles directly affect the induction of cellular and
humoral immunity [148].
It should be emphasized that GNPs are not biodegradable.
Therefore, the biodistribution and excretion kinetics have to
be studied comprehensively for different animal models. As
the excretion of accumulated particles from the liver and
spleen can take up to 3–4 months, the question as to the
injected doses and possible inflammation processes is still of
great importance. Bioaccumulated GNPs can interfere with
different diagnostic techniques, or accumulated GNPs can
exhibit catalytic properties. All these concerns, together with
potential toxicity, are big limitations of GNPs on a successfully
clinical translation. Nowadays, despite the huge numbers of
studies regarding the synthesis and functionalization of GNPs
(different shapes, coatings, sizes, charges, etc.), there are very
few nanomaterial-based pharmaceuticals on the market.
Figure 5. Mechanisms of immune system activation by engineered nanomaterials [11].
EXPERT REVIEW OF VACCINES 9
The synthetic and natural polymeric biodegradable nanoma-
terials also can serve as antigen carriers. The advantages of
nanoparticles of this type are that they are utilized well in the
organism, the target substance is highly efficiently involved, there
is a higher capability to overcome different physiological barriers,
and there are less systematic side effects. The action mechanisms
of biodegradable nanoparticles and GNPs as carriers of antigens
intheimmunesystemareprobablysimilar.Thesetwotypesof
nanoobjects can compete in the development of a new genera-
tion of vaccines, taking into account data on bioinertness, low
toxicity, and the good excretion of gold nanoparticles from the
organism with the involvement of a hepatobiliary system [147].
Thus, GNPs, which have adjuvant properties, can be an
excellent tool in the design of effective vaccines against infec-
tious diseases.
Acknowledgments
I thank Mr DN Tychinin for his help in preparation of the manuscript.
Funding
This work was supported by the Russian Science Foundation under grant
no. 19-14-00077.
Declaration of interest
The author has no relevant affiliations or financial involvement with any
organization or entity with a financial interest in or financial conflict with
the subject matter or materials discussed in the manuscript. This includes
employment, consultancies, honoraria, stock ownership or options, expert
testimony, grants or patents received or pending, or royalties.
Reviewer Disclosures
Peer reviewers on this manuscript have no relevant financial or other
relationships to disclose.
ORCID
Lev A. Dykman http://orcid.org/0000-0003-2440-6761
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EXPERT REVIEW OF VACCINES 13
