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Regulation of chemokine receptor CCR5 and production of RANTES and MIP-1α by interferon-β
Abstract
Trafficking of inflammatory T cells into the brain is associated with interactions of certain chemokines with their receptors, which plays an important role in the pathogenesis of multiple sclerosis (MS). We examined whether interferon-beta (IFN-beta) had the ability to regulate the production of chemokines and the expression of their receptors in T cells derived from patients with MS. It was demonstrated for the first time that in vitro exposure of T cells to IFN-beta-1a selectively inhibited mRNA expression for RANTES and MIP-1alpha and their receptor CCR5. T cell surface expression of CCR5 was significantly reduced in MS patients treated with IFN-beta, correlating with decreased T cell transmigration toward RANTES and MIP-1alpha. The study provides new evidence suggesting that IFN-beta treatment impairs chemokine-induced T cell trafficking by reducing the production of RANTES and MIP-1alpha and the expression of their receptors CCR5.
... Th1 # IFN-g production by MOG-primed splenocytes [51] # Th1 infiltration in CNS [53] # Expression of Th1 chemokines and their receptors on CD4 + T in the CNS [54] # IFN-g production by Th1 cells [53] # Expression of T-bet and IFN-g genes in the blood [43] # CCR5 mRNA levels and its ligands on T cells [56] " CCR5, IL-12Rb2 and IFN-g mRNA levels in ...
... Accordingly, the gene expression of transcription factor T-bet, as well as IFN-g, both Th1 cell-associated markers, was reduced in the blood from IFN-b treated MS patients [43]. Although in the aforementioned studies, the observed effect could be a result of IFN-b action on bystander cells, a direct effect on Th1 cells was previously demonstrated by Zang et al.; IFN-b treatment of T cells isolated from the blood of MS patients led to lower mRNA levels of CCR5 and its ligands, RANTES and MIP-1a compared to non IFN-b treated samples [56]. In the same study, similar results were reported in vivo upon IFN-b treatment of MS patients [56]. ...
... Although in the aforementioned studies, the observed effect could be a result of IFN-b action on bystander cells, a direct effect on Th1 cells was previously demonstrated by Zang et al.; IFN-b treatment of T cells isolated from the blood of MS patients led to lower mRNA levels of CCR5 and its ligands, RANTES and MIP-1a compared to non IFN-b treated samples [56]. In the same study, similar results were reported in vivo upon IFN-b treatment of MS patients [56]. On the other side, several studies suggest that the effects of IFN-b are not only anti-inflammatory but also pro-inflammatory. ...
... The results of RT-PCR analysis indicate that by days 6 through 8 post-infection, mRNA expression of RANTES is significantly up-regulated (p < 0.002) in infected mice compared with controls, indicating that it is involved in the immunopathogenesis in P. yoelii 17XL-infected mouse. RANTES in addition to CCR1 and CCR5 are expressed by Th1 cells [30]. Indeed, trafficking of inflammatory Th1 cells into the brain was reportedly mediated largely by RANTES interaction with CCR5 receptor [30]. ...
... RANTES in addition to CCR1 and CCR5 are expressed by Th1 cells [30]. Indeed, trafficking of inflammatory Th1 cells into the brain was reportedly mediated largely by RANTES interaction with CCR5 receptor [30]. Also the absence of CCR5 receptor in Plasmodium berghei ANKA-infected mouse brain resulted in a reduced Th1 cytokine production [9]. ...
... The expression of RANTES and CCR5 mRNA in P. yoelii 17XL-infected mouse brain in this study suggests a Th1-mediated immune response, and that factors capable of inducing Th1 response could play an important role in modulating malaria infections. Macrophages and other leukocytes release proinflammatory cytokines, including TNF-alpha, IFN-gamma and IL-1beta, which in turn will promote the release of chemokines [30]. The expression of chemokine, IP-10 and MCP-1, genes in KT-5, an astrocyte cell line, have been shown to be upregulated in vitro upon stimulation with a crude antigen of malaria parasites [12]. . ...
Abstract
Background
Malaria afflicts 300–500 million people causing over 1 million deaths globally per year. The immunopathogenesis of malaria is mediated partly by co mplex cellular and immunomodulator interactions involving co-regulators such as cytokines and adhesion molecules. However, the role of chemokines and their receptors in malaria immunopathology remains unclear. RANTES (Regulated on Activation Normal T-Cell Expressed and Secreted) is a chemokine involved in the generation of inflammatory infiltrates. Recent studies indicate that the degradation of cell-cell junctions, blood-brain barrier dysfunction, recruitment of leukocytes and Plasmodium -infected erythrocytes into and occlusion of microvessels relevant to malaria pathogenesis are associated with RANTES expression. Additionally, activated lymphocytes, platelets and endothelial cells release large quantities of RANTES, thus suggesting a unique role for RANTES in the generation and maintenance of the malaria-induced inflammatory response. The hypothesis of this study is that RANTES and its corresponding receptors (CCR1, CCR3 and CCR5) modulate malaria immunopathogenesis. A murine malaria model was utilized to evaluate the role of this chemokine and its receptors in malaria.
Methods
The alterations in immunomodulator gene expression in brains of Plasmodium yoelii 17XL-infected mice was analysed using cDNA microarray screening, followed by a temporal comparison of mRNA and protein expression of RANTES and its corresponding receptors by qRT-PCR and Western blot analysis, respectively. Plasma RANTES levels was determined by ELISA and ultrastructural studies of brain sections from infected and uninfected mice was conducted.
Results
RANTES (p < 0.002), CCR1 (p < 0.036), CCR3 (p < 0.033), and CCR5 (p < 0.026) mRNA were significantly upregulated at peak parasitaemia and remained high thereafter in the experimental mouse model. RANTES protein in the brain of infected mice was upregulated (p < 0.034) compared with controls. RANTES plasma levels were significantly upregulated; two to three fold in infected mice compared with controls (p < 0.026). Some d istal microvascular endothelium in infected cerebellum appeared degraded, but remained intact in controls.
Conclusion
The upregulation of RANTES, CCR1, CCR3, and CCR5 mRNA, and RANTES protein mediate inflammation and cellular degradation in the cerebellum during P. yoelii 17XL malaria.
... 63,64 It is speculated that IFN-β negatively affects the CCR5 signaling axis involving IFN-γ-secreting Th1 type T cells that respond to ligands macrophage inflammatory protein 1 alpha and CCL5 on macrophages, microglia, and astrocytes to promote immune cell recruitment to the brain. 65,66 Furthermore, IFN-β inhibits T-cell secretion of MMP-9, which degrades extracellular matrix and facilitates T-cell movement from the glia limitans into the brain parenchyma. 66 Hence, while in the case of MS, treatment with recombinant IFN-β may be useful to contain autoimmunity, for GBM where IFN-β is known to be upregulated, treatment with IFN-β inhibitors Neuro-Oncology may be useful to increase T-cell trafficking and activation in the CNS. ...
... 65,66 Furthermore, IFN-β inhibits T-cell secretion of MMP-9, which degrades extracellular matrix and facilitates T-cell movement from the glia limitans into the brain parenchyma. 66 Hence, while in the case of MS, treatment with recombinant IFN-β may be useful to contain autoimmunity, for GBM where IFN-β is known to be upregulated, treatment with IFN-β inhibitors Neuro-Oncology may be useful to increase T-cell trafficking and activation in the CNS. ...
Glioblastoma (GBM) is a highly malignant CNS tumor with very poor survival despite intervention with conventional therapeutic strategies. Although the CNS is separated from the immune system by the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB), emerging evidence of immune surveillance and the selective infiltration of GBMs by immune suppressive cells indicates that there is breakdown or compromise of these physical barriers. This in turn offers hope that immunotherapy can be applied to specifically target and reduce tumor burden. One of the major setbacks in translating immunotherapy strategies is the hostile microenvironment of the tumor that inhibits trafficking of effector immune cells such as cytotoxic T lymphocytes into the CNS. Incorporating important findings from autoimmune disorders such as multiple sclerosis to understand and thereby enhance cytotoxic lymphocyte infiltration into GBM could augment immunotherapy strategies to treat this disease. However, although these therapies are designed to evoke a potent immune response, limited space in the brain and cranial vault reduces tolerance for immune therapy induced inflammation and resultant brain edema. Therefore, successful immunotherapy requires that a delicate balance be maintained between activating and retaining lasting anti-tumor immunity.
... The ability of peripheral T cells migrate to the CNS through BBB is related to the expression of chemokines and chemokine receptors (Zang et al., 2000). CCR5 is a chemokine receptor regulated upon activation, showing potential association with MS, the overexpression of CCR5 may be associated with the influx of proinflammatory T cells into the CNS (Iarlori et al., 2000;Zang et al., 2001). RANTES, also named CCL5, is the principal chemotactic agent that induces recruitment of immune cells to the CNS. ...
... The decreasing of CCR5 expression in circulating γδ T cells of RRMS patients can be a result of IFN-β treatment, since it is capable of decrease RANTES production in CNS (Trebst et al., 2001) as well as the expression of CCR5 in the periphery, either in vitro or in vivo (Zang et al., 2001;Cheng et al., 2015). ...
We characterized circulating gamma-delta T cells in relapsing-remitting multiple sclerosis (RRMS) patients, during remission and relapse phases. In relapse, we observed a decrease of circulating CCR5+γδ TEMRAcell subset, together with a decrease in EOMES and granzyme B mRNA expression in γδ T cells, suggesting a reduction of the cytotoxic potential of this subset. Moreover, we also found a higher frequency of IFNγ+γδ T cells, which may indicate that these cells are assuming a more regulatory function associated to a Th1 profile. These results suggest a specific release from the periphery of a particular γδ T cell subset, expressing CCR5 and belonging to an effector compartment, supporting the idea that γδ T cells could play a role in MS relapse.
... IFN-¯ is currently a common treatment modality for MS. A recent study has shown that in vitro exposure of T cells to IFN-¯1a selectively inhibited mRNA expression for CCL5 and CCL3 (Zang et al, 2001 ). Furthermore, T-cell surface expression of CCR5 was signicantly reduced in MS patients treated with IFN-¯, correlating with decreased T-cell transmigration toward CCL5 and CCL3 (Zang et al, 2001 ). ...
... A recent study has shown that in vitro exposure of T cells to IFN-¯1a selectively inhibited mRNA expression for CCL5 and CCL3 (Zang et al, 2001 ). Furthermore, T-cell surface expression of CCR5 was signicantly reduced in MS patients treated with IFN-¯, correlating with decreased T-cell transmigration toward CCL5 and CCL3 (Zang et al, 2001 ). Collectively, these results demonstrate the emerging signicance of chemokine expression in the CNS during human demyelinating disease as well as the increase in chemokine receptor-bearin g cells and suggest a critical role for these molecules in the pathogenesis of disease development and progression. ...
Chemokines and their receptors are large families of inflammatory molecules responsible for a number of biologic functions including the accumulation of leukocytes at tissue sites. Over the past 8 years, a number of studies have indicated a role for chemokines in the pathogenesis of CNS inflammatory diseases. This minireview provides a brief summary of our current knowledge of chemokines and CNS inflammatory diseases including experimental autoimmune encephalomyelitis, multiple sclerosis, virus-induced demyelinating diseases, Alzheimer's disease, and central nervous system bacterial-induced diseases.
... We calculated a linear regression line between the channel intensities of the reporters for Ccl3 transcription and translation and found that its slope was reduced when IFN-I signaling was blocked. IFN-I is known to cause Ccl3 production (Bug et al., 1998;Zang et al., 2001;Salazar-Mather et al., 2002), and our study reveals that it does so primarily on the posttranscriptional level. Furthermore, we noted that the slope of the regression curve increased with time in organs of viral replication. ...
Chemokines guide immune cells during their response against pathogens and tumors. Various techniques exist to determine chemokine production, but none to identify cells that directly sense chemokines in vivo. We have generated CCL3-EASER (ErAse, SEnd, Receive) mice that simultaneously report for Ccl3 transcription and translation, allow identifying Ccl3-sensing cells, and permit inducible deletion of Ccl3-producing cells. We infected these mice with murine cytomegalovirus (mCMV), where Ccl3 and NK cells are critical defense mediators. We found that NK cells transcribed Ccl3 already in homeostasis, but Ccl3 translation required type I interferon signaling in infected organs during early infection. NK cells were both the principal Ccl3 producers and sensors of Ccl3, indicating auto/paracrine communication that amplified NK cell response, and this was essential for the early defense against mCMV. CCL3-EASER mice represent the prototype of a new class of dual fluorescence reporter mice for analyzing cellular communication via chemokines, which may be applied also to other chemokines and disease models.
... The increase in cellular vulnerability occurs by disinhibiting the expression of viral co-receptors, CCR5 and CXCR4. The latter mechanism occurs under physiologic conditions [80][81][82][83], but not in artificially stimulated cells [84]. In conjunction with immune activation (e.g., via proinflammatory cytokines or ligation of the T cell receptor), these factors facilitate viral replication, and thereby, accelerate disease pathogenesis. ...
The immune and sympathetic nervous systems are major targets of human, murine and simian immunodeficiency viruses (HIV-1, MAIDS, and SIV, respectively). The spleen is a major reservoir for these retroviruses, providing a sanctuary for persistent infection of myeloid cells in the white and red pulps. This is despite the fact that circulating HIV-1 levels remain undetectable in infected patients receiving combined antiretroviral therapy. These viruses sequester in immune organs, preventing effective cures. The spleen remains understudied in its role in HIV-1 pathogenesis, despite it hosting a quarter of the body’s lymphocytes and diverse macrophage populations targeted by HIV-1. HIV-1 infection reduces the white pulp, and induces perivascular hyalinization, vascular dysfunction, tissue infarction, and chronic inflammation characterized by activated epithelial-like macrophages. LP-BM5, the retrovirus that induces MAIDS, is a well-established model of AIDS. Immune pathology in MAIDs is similar to SIV and HIV-1 infection. As in SIV and HIV, MAIDS markedly changes splenic architecture, and causes sympathetic dysfunction, contributing to inflammation and immune dysfunction. In MAIDs, SIV, and HIV, the viruses commandeer splenic macrophages for their replication, and shift macrophages to an M2 phenotype. Additionally, in plasmacytoid dendritic cells, HIV-1 blocks sympathetic augmentation of interferon-β (IFN-β) transcription, which promotes viral replication. Here, we review viral–sympathetic interactions in innate immunity and pathophysiology in the spleen in HIV-1 and relevant models. The situation remains that research in this area is still sparse and original hypotheses proposed largely remain unanswered.
... 101 Treatment with IFN-β reduced the expression of CCR5 on T cells of MS patients, whereas treatment of T cells with IFN-β in vitro, inhibited their expression of this chemokine receptor as well its ligands CCL5 and CCL3. 102 Furthermore, the expression of CXCR3 on CD4 + and CD8 + T cells was considerably decreased in RRMS patients after treatment with IFN-β. 103 However, IFN-β induced a transient strong increase of CXCL10 level in MS patients. ...
Multiple sclerosis (MS) is an immune-mediated and neurodegenerative disorder that results in inflammation and demyelination of the central nervous system (CNS). MS symptoms include walking difficulties, visual weakening, as well as learning and memory impairment, thus affecting the quality of the patient’s life. Chemokines and chemokine receptors are expressed on the immune cells as well as the CNS resident cells. Several sets of chemokine receptors and their ligands tend to be pathogenic players in MS, including CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL17, CCL19, CCL21, CCL22, CXCL1, CXCL8, CXCL9, CXCL10, CXCL11, and CXCL16. Furthermore, current modulatory drugs that are used in the treatment of MS and its animal model, the experimental autoimmune encephalomyelitis (EAE), affect the expression of several chemokine and chemokine receptors. In this review, we highlight the pathogenic roles of chemokines and their receptors as well as utilizing them as potential therapeutic targets through selective agents, such as specific antibodies and receptor blockers, or indirectly through MS or EAE immunomodulatory drugs.
... The hepatitis C virus induces RANTES production by macrophages 46 . Evidence suggests that IFN type I inhibits the production of RANTES by T cells 47 . Therefore, consistent with the expected results, RANTES levels did not increase during TT treatment in the SVR group ( Table 3). ...
Abstract INTRODUCTION: Chronic hepatitis C is a leading cause of liver disease. Infection triggers an immediate immune response in the host that is mediated by humoral/cellular mechanisms. T cells respond to infection via secretion of cytokines, which inhibit or stimulate one another, leading to cytokine imbalance and ultimately affecting treatment. Studies using interferon (IFN) and ribavirin (RBV) showed that TCD8+ cells and cytokine levels are associated with sustainable virological response (SVR). However, studies that investigated the effects of triple therapy (TT) are limited. METHODS: The study included hepatitis C virus (HCV)+ RNA, naives, genotype 1, ≥18 years, and advanced fibrosis (F≥3) patients. Samples were collected at baseline and after 12 weeks (W12) of TT. Six cytokines were analyzed by flow cytometry. RESULTS: Of 31 patients, four were excluded (two deaths, one interrupted TT, and one F2 patient). Of the 27 remaining patients, 21 (78%) were cirrhotic. SVR was achieved in 63% of the patients. The patients had a mean age of 55.11 ± 10.03 years. Analyses at baseline showed that the chemokine CCL5/Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES) (p=0.04) and interleukin (IL)-6 (p=0.02), which was associated with SVR. RANTES (p=0.04) and IL-8 (p=0.01) levels were associated with SVR at W12. CONCLUSIONS Similar to patterns observed during double therapy, IL-6, IL-8, and RANTES levels were associated with SVR in TT, indicating the potential role of interferon in immune response to hepatitis C virus.
... 220 IFN-β-1a treatment decreased the frequency of CD4 1 CCR5 1 T-cells in peripheral blood and increased the levels of CCL2 in serum. 221,222 These clinical findings suggest that the chemokines and their receptors have been attractive targets for MS treatment. Clinical trials have been conducted using the following CCR2 antagonists or CCR2 neutralizing antibody: MK-0812, INCB8696, CCX915, and MLN1202. ...
... In vitro treatment of T cells with IFN β-1a inhibits expression of CCL3, CCL5 and CCR5 at mRNA level. IFN-β treatment decreases levels of CCR5 + CD4 + T cells, and CCL3, CCL4 and CCL5 in brain and spinal cord resulting in inhibition of EAE severity that is associated with reduction of induced-T cell migration toward CCL3 and CCL5 into the CNS by prevention of p38-MAPK and ERK1/2 activation [69,90]. However, expression of CCR5 on peripheral blood mononuclear cells is up-regulated in MS patients following 6 months of treatment with IFN β-1b [91]. ...
Background & objective:
Multiple sclerosis is an autoimmune demyelinating disease of the human central nervous system with still unknown etiology. Infiltration, accumulation and activation of autoreactive T cells, macrophages and other inflammatory immune cells in the CNS are the crucial steps in MS neuropathogenesis and development. Chemokines and their receptors play the main role in the attraction of the pathogenic cells into the CNS in MS. Specific chemokines and chemokine receptors are up-regulated in the actively demyelinating lesions and cerebrospinal fluid of MS patients. Many medical studies investigated how changes in levels or activities of chemokines and their receptors are implicated in leukocyte migration into CNS and consequently causing MS. These chemokines and their receptors are under intense focus to introduce new therapeutic strategies for MS.
Conclusion:
The aim of this review is to summarize previous findings on the relationship between chemokines network and MS development. Furthermore, opportunities and challenges in the chemokine system intervention as a potential therapeutic target for the treatment of MS will be outlined.
... Antiinflamatorni efekti interferona-β ostvaruju se sprečavanjem migracije Th1 limfocita u tkivo CNS. Interferon-β može da smanji ekspresiju hemokina i hemokinskih receptora karakterističnih za Th1 limfocite, kao što su CCL5, CCL3 i CXCR3, pri čemu se povećava ekspresija receptora tipičnog za Th2 limfocite, CCR4 (13,95). Glatiramer acetat je kopolimer sastavljen od aminokiselina koje se sa visokom efikasnošću vezuju za HLA klase II, a sa niskim afinitetom za receptore Th1 limfocita, prevodeći Th1 u Th2 ćelijski odgovor; pri tome rastu nivoi citokina koje produkuju Th2 limfociti, dok opadaju nivoi citokina koje sekretuju Th1 limfociti i ekspresija Th1-tipičnih hemokinskih receptora, CCR5 i CXCR3 (96,97). ...
Multiple sclerosis (MS) is a chronic inflammatory and autoimmune, demyelinating disease of the central nervous system (CNS). Migration of autoreactive T lymphocytes into the CNS is one of the key processes that characterize the pathogenesis of MS and lead to the formation of inflammatory demyelinating CNS lesions. Chemokines represent a family of cytokines. They are low molecular weight soluble proteins, which affect target cells by binding to membrane chemokine receptors. Previous studies on experimental model of MS and experimental clinical studies confirmed that numerous chemokines and chemokine receptors, synthesized by CNS cells and various leukocyte populations, are an important component in the pathogenesis of MS. The essential role of chemokines in MS pathogenesis is to stimulate the migration of peripheral leukocytes into the CNS. In clinical pharmacological studies, it was demonstrated that the impact on expression of chemokines and chemokine receptors represents one of the mechanisms of action of pharmacological immunomodulatory agents used in the treatment of MS. It is difficult to allocate a pharmacological agent that permanently effectively suppresses the disease, and one of the reasons for that is complexity of the network of chemokines and chemokine receptors, in terms of their multitude and their variety of functions. Improved therapeutics that would be produced in the future, by acting through the individual components of complex network of chemokines and chemokine receptors, would selectively influence the migratory properties of pathogenic leukocyte populations, without compromising the functions of other components of the immune system.
... The first to be approved were beta interferons (IFN-β-1a and IFN-β-1b), which have pleiotropic effects on cellular functions and may prevent leukocyte migration across the blood brain barrier by altering the expression of adhesion molecules as well as Glatiramer acetate, a MHC-binding protein that engages the T cell receptor (TCR) (Kozovska et al., 1999;Zang et al., 2001;Muraro et al., 2004). These agents are generally well tolerated and moderately effective in the short term in preventing relapses and attenuation of lesion activity visible on magnetic resonance imaging (Galetta et al., 2002;Cohen and Rivera, 2010). ...
... Second, glatiramer acetate (GA), formerly known as copolymer 1, is widely used for treatment of MS via increasing the levels of Th2-related cytokines and CCR7 expression, decreasing Th1-related cytokines as well as the expression of CCR5, CXCR3, and CXCR6 on T cells [40,76,[81][82][83][84]. Third, IFN-has been proven as an effective drug for treatment of RRMS patients for many years [85][86][87]. Dhib-Jalbut et al. demonstrated that IFN-treatment could reduce the expression of CCL5, CCL3, and CXCR3, which was associated with Th1 cells, increasing the expression of CCR4 which was often expressed by Th2 cells in MS patients [88,89]. ...
Multiple sclerosis is an autoimmune disease with classical traits of demyelination, axonal damage, and neurodegeneration. The migration of autoimmune T cells and macrophages from blood to central nervous system as well as the destruction of blood brain barrier are thought to be the major processes in the development of this disease. Chemokines, which are small peptide mediators, can attract pathogenic cells to the sites of inflammation. Each helper T cell subset expresses different chemokine receptors so as to exert their different functions in the pathogenesis of MS. Recently published results have shown that the levels of some chemokines and chemokine receptors are increased in blood and cerebrospinal fluid of MS patients. This review describes the advanced researches on the role of chemokines and chemokine receptors in the development of MS and discusses the potential therapy of this disease targeting the chemokine network.
... Figure 6 provides a theoretical model that integrates existing results from human clinical studies relating SNS activity to indicators of HIV-1 pathogenesis (Ironson et al., in press), in vitro analyses of catecholamine effects on HIV-1 replication (Cole et al., 1998;Cole et al., 1999;Collado et al., 2006;), and experimental analyses of the SIV lymphoid tissue model in vivo . SNS activity is hypothesized to enhance viral replication by inhibiting the activity of Type I interferons (Collado et al., 2006;), which increases viral replication through multiple mechanisms including impaired resistance to viral gene expression (via inhibition of the interferon-mediated antiviral state) (Collado et al., 2006) and enhanced cellular vulnerability to infection (via disinhibited expression of the viral coreceptors CCR5 and CXCR4, which occurs under physiologic conditions (Zang et al., 2001), but not in artificially stimulated cells (Yang et al., 2001). In conjunction with immune activation (e.g., via proinflammatory cytokines or ligation of the T cell receptor), these factors. ...
Chronic Psychological stress (CPS) has several adverse effects both on HIV people and on HIV+patient. HIV-people with CPS are more susceptible to HIV infection than the HIV-
people without CPS. T-cells have CXCR4 receptor and Macrophages have CCR5 receptor which can bind with both glucocorticoid and catecholamine hormone. HIV has GP120 residue which is able to bind with both CD4 and CXCR4/CCR5 receptor for its entry into the host cells. CPS increases glucocorticoid and catecholamine concentration in blood and thereby activates cAMP signaling pathway through binding the CXCR4/CCR5 receptors expressed on T cells and macrophages. This signal transduction pathway leads
to the synthesis of more CXCR4 and CCR5 receptors by those cells, and in turn the cells become more susceptible to HIV infection. On HIV+patient stress hormones arrest the infected cell in G2 phase which is favorable for HIV replication. At the same time T-cells and macrophage are more susceptible to HIV, so HIV can infects a lot of immune cells and thereby makes the immune system weak. Stress also inhibits Th2 when the cell produces INF-γ as a response to viral attack. So that other cells remain vulnerable to viral infection. When T-cell count is decreased in the blood, the body cannot protect itself from other opportunistic infectious pathogens. As a result progression of AIDS increases rapidly.
... reported as a main effect of IFN-α by several authors919293949596 . Although different findings have been published regarding the impact of IFN-α on CCL5 gene expression in a variety of cell lines979899100 , there is evidence that type I IFNs have the ability to reduce CCL5 production in human T cells [101] or other cell types [99, 100]. In line with these observations is the suppression of initially increased CCL5 mRNA levels in PB from HCV-infected patients with EF, measured 12 h after the first administration of Peg-IFN-α [16]. ...
C-C motif ligand 5 (CCL5) facilitates induction of chemotaxis in immune cells and activation of hepatic stellate cells (HSC) at sites of liver inflammation during chronic hepatitis C virus (HCV) infection. Importantly, CCL5 participates in the establishment of T-helper 1 responses crucial in controlling liver disease and HCV infection outcome and demonstrates distinct gene expression patterns between the blood and the liver, stressing the importance of immunoregulatory networks differentially functioning between these compartments. This review illustrates the significance of CCL5-dependent pathways in HCV-related immunopathogenesis by elaborating on biological mechanisms interconnecting peripheral and tissue immunology, liver pathology, HSC activation, and interferon-α immunotherapy.
... RANTES/CCL5 and antibodies against RANTES/CCL5 were reconstituted in sterilized artificial fluid (CMA/ microdialysis AB, Stockholm, Sweden). Anti-RANTES/CCL5 was injected at a dose of 2.0 μg/μl, which is considered enough to specifically neutralize this chemokine as determined in vitro (Alkhatib et al., 1996; Zang et al., 2001) and in vivo (Tavares and Minano, 2000 ). The coldwater tail-flick test was used to measure the nociceptive response (Pizziketti et al., 1985). ...
The present data provide the first in vivo evidence that the proinflammatory chemokine, Regulated on Activation Normal T cell Expressed and Secreted (RANTES/CCL5) microinjected directly into the periaqueductal grey in rats, a brain region critical to the processing of pain signals, and a primary site of action of many analgesic compounds, induced hyperalgesia. Pretreatment with antibodies against RANTES/CCL5 prevented the hyperalgesic response, indicating that RANTES/CCL5 is able to interfere with the control of hyperalgesia at the level of the periaqueductal grey and suggesting that chemokine blockers could have analgesic properties.
... Several studies have shown that IFN-β reduces the migration of inflammatory cells across the blood-brain barrier [12]. This is likely accomplished by reducing the expression of endothelial adhesion molecules, ICAM-1 and VCAM-1, and by downregulating the production of chemokines [23,24]. Although the immunomodulatory effects of IFN-β have been described in brain-related diseases including multiple sclerosis and focal cerebral ischemia, we are the first to describe the modulatory effects of IFN-β in pulmonary inflammation. ...
Aneurysmal subarachnoid hemorrhage (SAH) affects relatively young people and carries a poor prognosis with a case fatality rate of 35%. One of the major systemic complications associated with SAH is acute lung injury (ALI) which occurs in up to one-third of the patients and is associated with poor outcome. ALI in SAH may be predisposed by neurogenic pulmonary edema (NPE) and inflammatory mediators. The objective of this study was to assess the immunomodulatory effects of interferon-β (IFN-β) on inflammatory mediators in the lung after experimental SAH.
Male Wistar rats were subjected to the induction of SAH by means of the endovascular filament method. Sham-animals underwent sham-surgery. Rats received IFN-β for four consecutive days starting at two hours after SAH induction. After seven days, lungs were analyzed for the expression of inflammatory markers.
SAH induced the influx of neutrophils into the lung, and enhanced expression of the pulmonary adhesion molecules E-selectin, inter-cellular adhesion molecule (ICAM)-1, and vascular cell adhesion molecule (VCAM)-1 compared to sham-animals. In addition, SAH increased the expression of the chemokines macrophage inflammatory protein (MIP)-1α, MIP-2, and cytokine-induced neutrophil chemoattractant (CINC)-1 in the lung. Finally, tumor necrosis factor-α (TNF-α) was significantly increased in lungs from SAH-animals compared to sham-animals. IFN-β effectively abolished the SAH-induced expression of all pro-inflammatory mediators in the lung.
IFN-β strongly reduces lung inflammation after experimental SAH and may therefore be an effective drug to prevent SAH-mediated lung injury.
... This effect was statistically significant for CCL3. It was shown earlier that IFN-beta inhibited CCL3 and CCL5 gene expression in T cells (Zang et al., 2001) but induced CCL3, CCL4 and CCL5 chemokines in microglia cells (Kitai et al., 2000). ...
Plasmacytoid dendritic cells (pDCs) are present in peripheral blood, leptomeninges and demyelinating lesions in patients with multiple sclerosis (MS). The ability of pDCs to produce chemokines and express the chemokine receptor CCR7 in MS is not known. We studied pDCs in MS patients and healthy subjects. The ability of pDCs to up-regulate CCR7 was significantly increased in untreated MS patients as compared to healthy subjects. IFN-beta treatment significantly inhibited TLR9 agonist-specific secretion of chemokines, which are ligands for CCR5-positive Th1 cells (CCL3, CCL4, and CCL5), and impaired TLR9 agonist-induced up-regulation of CCR7 and IFN-alpha in MS patients. This finding represents a new immunomodulatory effect of IFN-beta in patients with multiple sclerosis.
... In addition they upregulate adhesion molecule expression which will assist in transendothelial migration of autoreactive immune cells through the blood brain barrier (BBB) [22]. IL-8 [23,24], IP-10 [25], MCP-1 [26,27], MIP-1a [28], and MIG [29] levels were previously found to be differentially expressed in sera from patients with IFNb treated MS as compared to untreated MS patients. Although changes in serum Eotaxin are not known to be directly associated with IFNb treatment, it is regulated by Th2 cell mediated cytokines [30], which play a significant role in the IFNb treatment mechanism [31] is also found to be reduced in CSF from people with MS as compared to HC [32]. ...
Interferon beta (IFNbeta) is the most common immunomodulatory treatment for relapsing-remitting multiple sclerosis (RRMS). However, some patients fail to respond to treatment. In this study, we identified putative clinical response markers in the serum and plasma of people with multiple sclerosis (MS) treated with IFNbeta. In a discovery-driven approach, we use 2D-difference gel electrophoresis (DIGE) to identify putative clinical response markers and apply power calculations to identify the sample size required to further validate those markers. In the process we have optimized a DIGE protocol for plasma to obtain cost effective and high resolution gels for effective spot comparison. APOA1, A2M, and FIBB were identified as putative clinical response markers. Power calculations showed that the current DIGE experiment requires a minimum of 10 samples from each group to be confident of 1.5 fold difference at the p<0.05 significance level. In a complementary targeted approach, Cytometric Beadarray (CBA) analysis showed no significant difference in the serum concentration of IL-6, IL-8, MIG, Eotaxin, IP-10, MCP-1, and MIP-1alpha, between clinical responders and non-responders, despite the association of these proteins with IFNbeta treatment in MS.
... The majority of the immunomodulatory mechanisms of IFN-β are well characterized: IFN-β inhibits the proliferation of T lymphocytes, reduces the production of proinflammatory cytokines, such as IFN-α and interleukin-12, and increases interleukin-10 levels, therefore shifting the cytokine profile from an inflammatory Th1 response to a favorable Th2 type (Noronha et al., 1993;Karp et al., 2001). In addition, IFN-β was found to modulate the expression of other cytokines mRNA pro-and antiinflammatory regulatory genes, major histocompatibility complex antigens, adhesion molecules, chemokine receptors and of interferon type 1 receptor, leading to a reduced extravasation of immune cells through the blood brain barrier and to a changed interplay between inflammation factors (Weinstock-Guttman et al., 2008;Reder et al., 2008;Miller et al., 1996;Satoh et al., 1995;Soilu-Hänninen et al., 2005;Zang et al., 2001;Sørensen and Sellebjerg, 2002;Dupont et al., 2002;Gilli et al., 2008). ...
We had reported that immune cells from relapsing remitting multiple sclerosis (RR-MS) patients secrete low levels of BDNF and that there is a defective regulation of its secretion via DC40. We now studied the effect of interferon-beta (IFN-beta1) on the secretion and regulation of BDNF from immune cells in patients with RR-MS. The PBMCs of IFN-beta1a treated RR-MS patients secreted higher BDNF levels vs. untreated patients. Anti CD40 mAb stimulation of PBMCs of IFN-beta1a treated patients upregulated the BDNF levels. There was no significant effect of CD40 stimulation on PBMCs of untreated patients. CD40(+) expression on CD14(+) cells was higher in IFN-beta treated patients vs. untreated patients. In vitro treatment with IFN-beta1a of PBMCs from healthy controls and untreated patients led to a significant increase in CD40 expression on CD14(+) cells in both groups. The addition of IFN-beta1a to CD40 stimulated PBMCs of untreated patients restored the up regulatory effect of CD40 stimulation on BDNF levels. Therefore, reduced BDNF secretion from PBMCs and defective regulation effect of CD40 stimulation on BDNF levels in untreated RR-MS are reversible by therapy with IFN-beta1a.
... In vitro experiments showed that CCL3 increases the transmigration of bone marrow-derived dendritic cells across brain microvessel endothelial monolayers (Zozulya et al. 2007). The functional involvement of CCL3 has been suggested in several CNS disorders, including multiple sclerosis (Balashov et al. 1999;Zang et al. 2001) and Wallerian degeneration (Perrin et al. 2005). Deletion of CCL3 retards neurodegeneration in Sandhoff disease mice (Wu and Proia 2004), and delays CNS demyelination without blood-brain barrier disruption during cuprizone intoxication (McMahon et al. 2001). ...
Microglia are implicated as a source of diverse proinflammatory factors in the CNS. Extracellular nucleotides are well known to be potent activators of glial cells and trigger the release of cytokines from microglia through purinergic receptors. However, little is known about the role of purinoceptors in microglial chemokine release. In this study, we found that high concentrations of ATP evoked release of CC-chemokine ligand 3 (CCL3)/macrophage inflammatory protein-1alpha from MG-5 cells, a mouse microglial cell line, and rapid up-regulation of CCL3 mRNA was elicited within 30 min of ATP stimulation. The release of CCL3 was also stimulated by 2'- and 3'-O-(4-benzoylbenzoyl) ATP, an agonist of P2X(7) receptors. Brilliant Blue G, an antagonist of P2X(7) receptors, strongly inhibited this ATP-induced CCL3 release. Similar pharmacological profile was observed in primary microglia. In MG-5 cells, ATP caused de-phosphorylation and nuclear translocation of the transcription factor nuclear factor of activated T cells (NFAT). ATP-induced NFAT de-phosphorylation was also dependent on P2X(7) receptor activation. Furthermore, ATP-induced CCL3 release and production were prevented by a selective inhibitor of NFAT. Taken together, the results of this study demonstrate an involvement of NFAT in the mechanism underlying P2X(7) receptor-mediated CCL3 release.
Background
Multiple sclerosis (MS) is a chronic debilitating disease characterized by inflammatory demyelination of the central nervous system. Grey matter (GM) lesions have been shown to be closely related to MS motor deficits and cognitive impairment. In this study, GM lesion-related genes for diagnosis and immune status in MS were investigated.
Methods
Gene Expression Omnibus (GEO) databases were utilized to analyze RNA-seq data for GM lesions in MS. Differentially expressed genes (DEGs) were identified. Weighted gene co-expression network analysis (WGCNA), least absolute shrinkage and selection operator (LASSO) algorithm and protein-protein interaction (PPI) network were used to screen related gene modules and candidate genes. The abundance of immune cell infiltration was analyzed by the CIBERSORT algorithm. Candidate genes with strong correlation with immune cell types were determined to be hub genes. A diagnosis model of nomogram was constructed based on the hub genes. Gene set enrichment analysis (GSEA) was performed to identify the biological functions of hub genes. Finally, an MS mouse model was induced to verify the expression levels of immune hub genes.
Results
Nine genes were identified by WGCNA, LASSO regression and PPI network. The infiltration of immune cells was significantly different between the MS and control groups. Four genes were identified as GM lesion-related hub genes. A reliable prediction model was established by nomogram and verified by calibration, decision curve analysis and receiver operating characteristic curves. GSEA indicated that the hub genes were mainly enriched in cell adhesion molecules, cytokine-cytokine receptor interaction and the JAK-STAT signaling pathway, etc .
Conclusions
TLR9, CCL5, CXCL8 and PDGFRB were identified as potential biomarkers for GM injury in MS. The effectively predicted diagnosis model will provide guidance for therapeutic intervention of MS.
Multiple sclerosis starts with increased migration of auto-reactive lymphocytes across the blood-brain barrier, resulting in persistent neurodegeneration. Clinical and epidemiological studies indicated upper respiratory viral infections are associated with clinical exacerbation of multiple sclerosis. However, so far there is no any direct evidence to support it. Using the experimental autoimmune encephalomyelitis mice as the model for multiple sclerosis, we demonstrated that mice experienced with influenza virus infection were unable to recover from experimental autoimmune encephalomyelitis with a long-term exacerbation. The exacerbated disease was due to more type I T cells, such as CD45highCD4⁺CD44high, CD45highCD4⁺CCR5⁺, CD45high IFNγ⁺CD4⁺, MOG35-55-specific IFNγ⁺CD4⁺ and influenza virus-specific IFNγ⁺CD4⁺ T cells, infiltrating central nervous system in mice with prior influenza virus infection. Influenza virus infection created a notable inflammatory environment in lung and mediastinal lymph node after influenza virus inoculation, suggesting the lung may constitute an inflammatory niche in which auto-aggressive T cells gain the capacity to enter CNS. Indeed, the early stage of EAE disease was accompanied by increased CCR5⁺CD4⁺, CXCR3⁺CD4⁺ T cell and MOG35-55 specific CD4⁺ T cells localized in the lung in influenza virus-infected mice. CCL5/CCR5 might mediate the infiltration of type I T cells into CNS during the disease development after influenza infection. Administration of CCR5 antagonist could significantly attenuate the exacerbated disease. Our study provided the evidence that the prior influenza virus infection may promote the type I T cells infiltration into the CNS, and subsequently cause a long-term exacerbation of experimental autoimmune encephalomyelitis.
Background: The molecules that provide access to activated T cells in the CNS, including chemokines, have been considered to be a crucial step in the pathogenesis of multiple sclerosis (MS). Aims: In this study, we investigated serial serum chemokine levels in patients with relapsing-remitting MS over 1 year and the association of these chemokine levels with treatment regimens, lesions on M RI and patients' characteristics. Methods: Serum CXCL9, CXCL 10, CCL2, CCL4 and CCL5 levels were evaluated using ELISA every 2 months for a year in 28 healthy controls and 28 MS patients during their treatment with interferon (IFN)-beta. Patients underwent M RI and were evaluated using the Expanded Disability Status Scale (EDSS) at the first and final evaluations. Results: CXCL 10 scrum levels were higher in MS patients compared with controls, were positively correlated with 12 lesions on MRI and were slightly increased during relapses. Treatment with IFN beta-la or IFN beta-1b was associated with increased CXCL10 levels when evaluated more than 36 hours after subcutaneous injection. The CXCL9 levels were higher after MS relapse. There was significant variability in CCL4 and CCL5 levels in the serial evaluations, associated with gender and treatment. CCL2 levels were higher in treated MS patients than healthy controls, particularly among those patients with a stable form of the disease. Conclusion: Serum is a feasible resource for searching for an immunological marker in MS. Peripheral chemokine levels correlated in different ways with IFN beta) therapy and with disease and patient characteristics. Clinical trial registration number: ISRCTN45526724.
Quantum dots (QDs) are nano-sized semiconductors. Previously, intratracheal instillation of QD705s induces persistent inflammation and remodeling in the mouse lung. Expression of interferon beta (IFN-β), involved in tissue remodeling, was induced in the mouse lung. The objective of this study was to understand the mechanism of QD705 induced interferon beta (IFN-β) expression. QD705-COOH and QD705-PEG increased IFN-β and IP-10 mRNA levels during day1 to 90 post-exposure in mouse lungs. QD705-COOH increased IFN-β expression via Toll/interleukin-1 receptor domain-containing adapter protein (TRIF) dependent Toll-like receptor (TLR) signaling pathways in macrophages RAW264.7. Silencing TRIF expression with siRNA or co-treatment with a TRIF inhibitor tremendously abolished QD705s-induced IFN-β expression. Co-treatment with a TLR4 inhibitor completely prevented IFN-β induction by QD705-COOH. QD705-COOH readily entered cells, and co-treatment with either inhibitors of endocytosis or intracellular TLRs prevented IFN-β induction. Thus, activation of the TRIF dependent TLRs pathway by promoting endocytosis of TLR4 is one of the mechanisms for immunomodulatory effects of nanoparticles.
Studies of chemokines and their receptors in multiple sclerosis (MS) have been, to date, descriptive. Much of the information that has been reported concerning chemokines and chemokine receptors in MS can be readily correlated to findings in animal models, including experimental autoimmune encephalomyelitis (EAE). Chemokine and chemokine receptor expression in MS have also been reported. Less information is available about the expression of chemokine receptors on monocytes in the blood and cerebrospinal fluid (CSF) of patients with MS. Investigators has also addressed the question whether patients with MS show preferential expression of chemokine receptors on circulating cells. In summary, the conceptual scheme of leukocyte trafficking into inflammatory lesions, that was derived in studies using the EAE animal model, may be applicable as well to the human disease MS. This situation provides a favorable circumstance for the evaluation of experimental anti-inflammatory strategies that are targeted to either production of chemokines or to the function of chemokines. In the complicated milieu of the MS lesion, actions of individual chemokines may promote destructive inflammation or recruitment of vital and protective regulatory cells or even tissue remodeling and repair processes. The final definitive test of theories about chemokine action in MS will be provided by the efficacy that emanates from blocking individual receptors.
Interferons (IFN) are subdivided into type I IFN (IFN-I, here synonymous with IFN-α/β), type II (IFN-γ) and type III IFN (IFN-III/IFN-λ) that reprogram nuclear gene expression through STATs 1 and 2 by forming STAT1 dimers (mainly IFN-γ) or the ISGF3 complex, a STAT1-STAT2-IRF9 heterotrimer (IFN-I and IFN-III). Dominant IFN activities in the immune system are to protect cells from viral replication and to activate macrophages for enhanced effector function. However, the impact of IFN and their STATs on the immune system stretches far beyond these activities and includes the control of inflammation. The goal of this review is to give an overview of the different facets of the inflammatory process that show regulatory input by IFN/STAT.
Chronic inflammatory demyelinating polyneuropathy (CIDP) is a group of idiopathic, acquired, immune-mediated inflammatory demyelinating diseases of the peripheral nervous system. A majority of patients with CIDP respond to "first-line" treatment with IVIG, plasmapheresis and/or corticosteroids. There exists insufficient evidence to ascertain the benefit of treatment with "conventional" immunosuppressive drugs. The inconsistent efficacy, long-term financial burden and health risks of non-specific immune altering therapy have drawn recurrent attention to the possible usefulness of a variety of biological agents that target key aspects in the CIDP immunopathogenic pathways. This review aims to give an updated account of the scientific rationale and potential use of biological therapeutics in patients with CIDP. No specific treatment recommendations are given. The discovery, development and application of biological markers by modern molecular diagnostic techniques may help identify drug-naïve or treatment-resistant CIDP patients most likely to respond to targeted immunotherapy.
Interferon-β is a remarkably pleiotropic molecule. Antiviral, pro- and antiproliferative, pro- and antiapoptotic, and complex immunoregulatory activities have all been described. The precise mechanism(s) that underlie the beneficial effects of interferon-β in multiple sclerosis remain poorly understood; this has hindered progress in the search for more effective therapies. An increasing body of literature supports the hypothesis that interferon-β-mediated changes in the production and activities of the immunoregulatory cytokines interleukin-12 and interleukin-10 are important to the therapeutic benefits of interferon-β in multiple sclerosis. These data are reviewed here.
Chemokines are small molecular-weight chemotactic cytokines that can be classified into four subfamilies based on the position
of the amino-terminal cysteines (1,2). The CXC chemokines can be further categorized based on the presence or absence of a glutamate-leucine-arginine (ELR) motif
in the amino terminus. Chemokines that possess the ELR motif are generally chemotactic for neutrophils and are angiogenic,
whereas the non-ELR CxC chemokines are chemotactic for activated T cells and are angiostatic (3). The CC family of chemokines are chemoattractant for a variety of cell types, including monocytes/macrophages, T lymphocytes,
basophils, eosinophils, and dendritic cells (4–6). The C family, lymphotactin, is chemotactic for T cells and natural killer (NK) cells (7) and the CX3C chemokine contains a chemokine domain attached to a membrane-bound mucin chain that produces a soluble chemoattractant
after proteolysis or mRNA processing (8). This chemokine is a chemoattractant for T cells, NK cells, and neutrophils (2). Chemokines and chemokine receptor-like molecules are also encoded by viruses and used in a variety of strategies to subvert
the immune response (9).
Multiple sclerosis (MS) is considered an immune-mediated, demyelinating, and degenerative disease of the central nervous system
(CNS). The immune-mediated pathogenesis is supported by the significant infiltration of leukocytes into the CNS parenchyma
and by the finding of a substantial similarity between active MS plaques and lesions in the CNS of animals with experimental
autoimmune encephalomyelitis (EAE); EAE is produced by an autoreactive T-cell response. The favorable response of many MS
patients to immunomodulatory drugs, including use of β interferons (IFNs) and glatiramer acetate to decrease the number of
relapses and reduce magnetic resonance imaging (MRI) disease activity (1–4), also corroborates the hypothesis of immune-mediated injury.
Interferons, despite their common name, comprise a group of cytokines with quite different molecular structures, cellular receptors, biological effects, functions and applications. We here review studies directed at defining the role played by these molecules when they are produced endogenously or administered exogenously in the course of experimental autoimmune encephalomyelitis (EAE), a model disease considered relevant for the pathogenesis of multiple sclerosis (MS).
Studies on the role of Type II interferon, i.e. interferon-γ (IFN-γ), are almost unanimously indicative of a beneficial role, though this is in contrast to clinical observations pertaining to the role of IFN-γ in MS. Possible explanations for this discrepancy are considered in this review.
Interferon-β (IFN-β), one of the Type I interferons, also exerts a beneficial effect in EAE. In this case there is good correspondence with clinical observations, and studies in the animal model have contributed to providing possible explanations for these beneficial effects.
Our review also covers a limited number of studies on the apparently beneficial effect of treatment with another Type I interferon, namely IFN-τ, originally discovered as a pregnancy recognition hormone in sheep.
Mechanisms causing liver fibrosis during chronic hepatitis C virus infection (cHCV) are not sufficiently understood. This study was aimed to identify biomarkers for early fibrosis (EF) and to investigate their potential role in cHCV-related fibrogenesis. To this end, peripheral whole blood (PB) samples from 36 patients with cHCV recruited from two independent cohorts were subjected to microarray analysis 12 h before initiation of peginterferon-alpha (Peg-IFN-α) and ribavirin therapy. Liver biopsies were evaluated using the Batts-Ludwig staging (BL-S) classification system for fibrosis. We showed that gene expression profiles (N = 8) distinguished between EF (BL-S: 0,1) and late fibrosis (LF; BL-S: 2,3,4) with 88.9% accuracy. Fibrosis-related functional annotations for chemokine-'C-C-motif'' ligand 5 (CCL5) provided foundation for focused investigation, and qRT-PCR confirmed that CCL5 mRNA levels (PB) reliably discriminate EF from LF (accuracy: 86.7%). Positive correlations (P < 0.05) with CCL5 mRNA levels and EF discovered gene expression profiles (PB) reflecting stable expression of IFN-α receptor 1, negative regulation of the MyD88-dependent toll-like receptor (TLR) pathway and decreased expression of TLR3 in vivo. Remarkably, Peg-IFN-α suppressed CCL5 mRNA levels (PB) in EF in vivo. These findings along with results from parallel in vitro investigation into the effect of IFN-α or poly I:C (TLR3-agonist) on CCL5 gene expression in hepatic stellate cells (HSC) attest to the multi-site involvement of these pathways in regulating fibrogenesis. In conclusion, we identified novel, reliable biomarkers for EF and exposed functional properties of the molecular network regulating CCL5 biosynthesis in peripheral or hepatic cell types with key roles in cHCV-related liver and/or immune pathogenesis.
The molecules that provide access to activated T cells in the CNS, including chemokines, have been considered to be a crucial step in the pathogenesis of multiple sclerosis (MS).
In this study, we investigated serial serum chemokine levels in patients with relapsing-remitting MS over 1 year and the association of these chemokine levels with treatment regimens, lesions on MRI and patients' characteristics.
Serum CXCL9, CXCL10, CCL2, CCL4 and CCL5 levels were evaluated using ELISA every 2 months for a year in 28 healthy controls and 28 MS patients during their treatment with interferon (IFN)-β. Patients underwent MRI and were evaluated using the Expanded Disability Status Scale (EDSS) at the first and final evaluations.
CXCL10 serum levels were higher in MS patients compared with controls, were positively correlated with T2 lesions on MRI and were slightly increased during relapses. Treatment with IFNβ-1a or IFNβ-1b was associated with increased CXCL10 levels when evaluated more than 36 hours after subcutaneous injection. The CXCL9 levels were higher after MS relapse. There was significant variability in CCL4 and CCL5 levels in the serial evaluations, associated with gender and treatment. CCL2 levels were higher in treated MS patients than healthy controls, particularly among those patients with a stable form of the disease.
Serum is a feasible resource for searching for an immunological marker in MS. Peripheral chemokine levels correlated in different ways with IFNβ therapy and with disease and patient characteristics. Clinical trial registration number: ISRCTN45526724.
The purinergic receptor family contains some of the most abundant receptors in living organisms. A growing body of evidence indicates that extracellular nucleotides play important roles in the regulation of neuronal and glial functions in the nervous system through purinergic receptors. Nucleotides are released from or leaked through nonexcitable cells and neurons during normal physiological and pathophysiological conditions. Ionotropic P2X and metabotropic P2Y purinergic receptors are expressed in the central nervous system (CNS), participate in the synaptic processes, and mediate intercellular communications between neuron and gila and between glia and other glia. Glial cells in the CNS are classified into astrocytes, oligodendrocytes, and microglia. Astrocytes express many types of purinergic receptors, which are integral to their activation. Astrocytes release adenosine triphosphate (ATP) as a "gliotransmitter" that allows communication with neurons, the vascular walls of capillaries, oligodendrocytes, and microglia. Oligodendrocytes are myelin-forming cells that construct insulating layers of myelin sheets around axons, and using purinergic receptor signaling for their development and for myelination. Microglia also express many types of purinergic receptors and are known to function as immunocompetent cells in the CNS. ATP and other nucleotides work as "warning molecules" especially by activating microglia in pathophysiological conditions. Studies on purinergic signaling could facilitate the development of novel therapeutic strategies for disorder of the CNS.
In the last two decades MS has changed from an idiopathic condition with only symptomatic treatments to a disease with better characterized pathophysiologic underpinnings and several treatments that modify its long-term course based on specific mechanisms of action. There are now several FDA approved options for therapy at the onset of disease, and discussions have begun on choosing the best treatment in individual patients and what option to choose next in patients who are failing their current treatment. Numerous studies have begun to highlight that the underlying pathology of CNS damage may be different in subsets of patients, raising the possibility that some may respond to a treatment with a mechanism of action that is targeted to ‘their’ MS. Trials are ongoing of numerous new agents with different mechanisms of action and some combination therapies. A better understanding of how each therapy works may guide decisions on initiating, combining or changing therapy in a more rational way to improve patient outcomes. Further knowledge of underlying mechanisms of disease in different patients with ‘the same’ disease may lead to more targeted therapies, as will biomarkers that predict clinical response to therapy. Studies of the effects of various agents used in MS reveal both overlapping and distinct mechanisms of actions that may be relevant to their efficacy and side effects in individual patients. However, it is important to remember that most agents are approved based on their reduction of MRI lesions and relapse rates over a short time frame. These measures only partially correlate with long-term disability, which may be the most relevant clinical outcome for people with MS. Fixed disability requires years to become apparent, and there is a lack of large studies of biomarkers for chronic outcomes. In addition, few large studies correlate response to therapy with cognitive outcomes, which are a major cause of chronic disability.
Multiple sclerosis (MS) is a chronic demyelinating disease of the CNS. The most common form of MS at onset, relapsing-remitting disease, is defined not by pathological features but by unpredictable periods of acute or subacute neurological worsening, followed by gradual improvement over weeks to months, often with residual neurological deficits. Evidence from serial magnetic resonance imaging studies in relapsing-remitting disease reveals significant inflammation, demyelination and axonal loss occurring both during and between relapses. Disease-modifying agents, such as interferon (IFN)-beta(1a), reduce the frequency of relapse by 30% in well established relapsing patients, reduce the risk of a second attack by 50% in high-risk patients, following a first attack, and reduce the number and volume of magnetic resonance imaging lesions. Intramuscular IFN-beta(1a) is effective in delaying disability progression and brain atrophy. The relationship between response to therapy and pathological subtype of MS is unknown. This review summarizes key findings of Phase III clinical trials, extension studies and postmarketing trials, demonstrating the efficacy and safety of intramuscular IFN-beta(1a). Results demonstrating the negative impact of anti-IFN-beta neutralizing antibodies on clinical efficacy are also addressed. Finally, expert commentary regarding the treatment of MS with IFN-beta therapy and future strategies to augment intramuscular IFN-beta(1a) efficacy by combination treatment with other agents is presented.
High-dose, high-frequency IFN beta-1a in multiple sclerosis (MS) can prevent lesion formation, decrease the frequency/severity of relapses and delay progression of disability, with a proven safety profile. Rates of non-adherence are high. There are drugs under investigation that may have greater efficacy and different safety profiles from existing therapies.
Evidence supporting the efficacy of IFN beta-1a, factors contributing to non-adherence, and strategies to combat non-adherence. It is hoped that these strategies, coupled with future advances in pharmacogenetics, might lead to better outcomes. The PubMed database was searched using the terms "multiple sclerosis" and "interferon beta-1a", for papers published between 1998 and 2010. Relevant manuscripts and pivotal papers from clinical trials were cited. Searches of abstracts from congresses were also performed to obtain recent findings.
An overview of early pivotal trials, comparative studies with other treatments, and recent studies assessing the development of this therapy.
Long-term treatment with IFN beta-1a has benefits in MS and a good safety profile. Although adherence outside of clinical trials can be poor, injection devices, better tolerated drug formulations and education regarding treatment expectations are some of the strategies employed to help patients to adhere to treatment in the hope of improving outcomes.
Multiple Sklerose (MS) ist eine chronisch-entzündliche Erkrankung des Zentralen Nervensystems mit deutlich ausgeprägten Autoimmunphänomenen. Das derzeit meistverwendete Therapeutikum zur Sekundärprophylaxe von Krankheitsschüben ist rekombinantes Interferon-β (IFN-β). Wirk- und Nebenwirkungsmechanismen des Medikaments werden bisher nur partiell verstanden. In der Pathogenese der MS spielt eine Familie chemotaktisch wirksamer Zytokine, der Chemokine, eine entscheidende Rolle. Ziel dieser Studie war zu untersuchen, ob IFN-β die systemischen Konzentrationen der Pathogenese-relevanten Chemokine CXCL10, CCL2 und außerdem des endogenen Pyrogens IL-6 verändert, und ob diese Veränderungen mit dem Auftreten grippeartiger Nebenwirkungen korrelieren. Zu diesem Zweck wurden bei 37 Patienten mit schubförmiger MS zu drei Zeitpunkten vor sowie 6 und 24 Stunden nach der Applikation von IFN-β die genannten Botenstoffe im Blut bestimmt. Parallel wurden subjektiv empfundene grippeartige Nebenwirkungen mit Hilfe eines standardisierten Fragebogens abgefragt, und die Körperkerntemperatur wurde gemessen. Als Kontrollen dienten gesunde Probanden, derzeit nicht immunmodulatorisch behandelte MS-Patienten und MS-Patienten unter Therapie mit Glatirameracetat. Nur bei den mit IFN-β behandelten Patienten zeigte sich nach 6 Stunden ein signifikanter transienter Anstieg der Konzentrationen von CXCL10, CCL2. Der Anstieg der Chemokinkonzentrationen korrelierte mit einem transienten IL-6-Anstieg und dem Auftreten grippeartiger Nebenwirkungen. Chemokine, unter denen sich zahlreiche starke endogene Pyrogene befinden, könnten somit für die häufig zu beobachtenden grippeartigen Nebenwirkungen mit verantwortlich sein. Die Ergebnisse werfen die weiterführende Frage auf, ob die beobachtete Chemokininduktion auch relevant für den therapeutischen Effekt von IFN- ist. Ob Chemokine sich erfolgreich als Biomarker zur Prädiktion des Therapieerfolgs einsetzen lassen, wird derzeit in einem weiterführenden Projekt untersucht. Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system with clear autoimmune phenomena. Recombinant Interferon-β (IFN-β) is currently the most widely used treatment to prevent relapses. The mechanisms and side effects of the drug are only partially understood. A family of chemotactical active cytokines, the chemokines, play a decisive role in the pathogenesis of MS. The aim of this study was to examine if IFN-β alters the systemic concentrations of CXCL10 and CCL2, two chemokines that are relevant in the pathogenesis, and of IL-6, an endogenous pyrogen. A further aim was to discover whether these concentrations correlate with the appearance of flu-like symptoms, a common adverse effect of IFN-β. 37 patients with relapsing-remitting MS were tested three times to measure the chemokine concentrations in their blood prior to IFN-β application, and again 6 and 24 hours after application. Concurrently, the occurrence of flu-like symptoms was recorded with the help of a standardized questionnaire and through body temperature measurements. The control groups consisted of healthy subjects, MS patients not receiving any treatment, and MS patients treated with glatiramer acetate. After 6 hours, only the MS patients treated with IFN-β showed a significant transient elevation in the concentrations of CXCL10 and CCL2. This elevation correlated with a transient increase in the IL-6 concentration and the appearance of flu-like symptoms. Among the chemokines there are many strong endogenous pyrogens, which might be responsible for the commonly observed, flu-like side effects of IFN-β. The results raise the question of whether the observed induction of chemokines is also relevant for the therapeutic effect of IFN-β. Whether chemokines can be used as biomarkers to predict therapeutic success is currently being explored in ongoing work built upon this study.
Interferon beta has been approved for the treatment of multiple sclerosis (MS). It is believed that immunomodulatory rather than antiviral activity of interferon beta is responsible for disease amelioration. The impact of interferon beta on the chemoattraction of immune cells has not been fully addressed.
To address the influence of interferon beta on the expression of chemokines and their receptors in a standardized setting.
The expression of 14 chemokines and 14 chemokine receptor genes was determined by quantitative real-time polymerase chain reaction from fresh blood samples.
Outpatient units in Germany. Patients Untreated and interferon beta-treated patients with MS who tested positive and negative for neutralizing antibodies (NABs) were recruited from August 24, 2006, through December 15, 2006, for the initial study and from March 12, 2007, through April 2, 2007, for the validation study.
Gene expression and serum chemokine protein levels.
CCL1, CCL2, CCL7, CXCL10, CXCL11, and CCR1 gene expression was strongly upregulated in interferon beta-treated, NAB-negative MS patients. In contrast, gene expression in interferon beta-treated, NAB-positive MS patients did not differ from untreated control donor individuals. Antibody titers inversely correlated with chemokine and chemokine receptor gene expression. Accordingly, serum chemokine protein levels of interferon beta-treated, NAB-negative MS patients were significantly higher than in untreated or interferon beta-treated, NAB-positive MS patients.
We demonstrate that interferon beta strongly upregulates a set of chemokines and CCR1 in peripheral immune cells. The peripheral upregulation of these chemokines may reduce the chemoattraction of immune cells to the central nervous system and thus add to the therapeutic effects of interferon beta.
Interferon-beta (IFNbeta) is an extra-cellular protein mediator of host defense and homeostasis. IFNbeta has well-established direct antiviral, antiproliferative and immunomodulatory properties. Recombinant IFNbeta is approved for the treatment of relapsing-remitting multiple sclerosis. The immunomodulatory effects of IFNbeta administration failed to demonstrate consistent benefit during treatment of various autoimmune neuromuscular diseases. Existing studies were flawed due to the often uncontrolled and unblinded nature of protocols, small patient numbers per study, the undetermined optimum dose and schedule of IFNbeta therapy, and the relatively brief periods of IFNbeta administration and clinical follow-up for mostly chronic inflammatory disorders. Additional, controlled, prospective studies are needed to definitely establish the full potential of this cytokine for this group of diseases. IFNbeta therapy may trigger autoantibody production, but only rarely clinically overt autoimmune disease. Anecdotal reports hint at the exceptional association between IFNbeta treatment and the induction or exacerbation of a variety of immune-mediated neuromuscular diseases, likely in genetically predisposed individuals. Thus, recombinant IFNbeta has the theoretical potential to either treat or cause autoimmune neuromuscular disorders by altering the complicated and delicate balances within the immune system networks.
There is at present no cure for multiple sclerosis (MS), and existing therapies are designed primarily to prevent lesion formation, decrease the rate and severity of relapses and delay the resulting disability by reducing levels of inflammation.
The aim of this review was to assess the treatment of relapsing MS with particular focus on subcutaneous (s.c.) interferon (IFN) beta-1a.
The literature on IFN beta-1a therapy of MS was reviewed based on a PubMed search (English-language publications from 1990) including its pharmacodynamics and pharmacokinetics, clinical efficacy in relapsing MS as shown in placebo-controlled studies and in comparative trials, efficacy in secondary progressive MS, safety and tolerability, and the impact of neutralizing antibodies.
The literature suggests that high-dose, high-frequency s.c. IFN beta-1a offers an effective option for treating patients with relapsing MS, with proven long-term safety and tolerability, and has a favourable benefit-to-risk ratio compared with other forms of IFN beta.
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS). Interferon (IFN) beta products are potent antiinflammatory and immunomodulatory agents that interfere with the autoimmune processes in MS and reduce CNS damage. Once-weekly intramuscular IFN beta-1a slows the progression of neurologic and cognitive disability, reduces the frequency of relapses and inflammatory lesion burden, and preserves cognitive function. It has a positive effect on both conventional and nonconventional measures of magnetic resonance imaging (MRI) and reduces the progression of brain atrophy, predominantly due to reduced grey matter atrophy. Early initiation of disease-modifying therapy after the diagnosis of relapsing-remitting MS or after a single demyelinating event (and evidence of lesions on MRI) allows patients the opportunity to obtain maximal long-term benefits.
Interferon-beta is a remarkably pleiotropic molecule. Antiviral, pro- and antiproliferative, pro- and antiapoptotic, and complex immunoregulatory activities have all been described. The precise mechanism(s) that underlie the beneficial effects of interferon-beta in multiple sclerosis remain poorly understood; this has hindered progress in the search for more effective therapies. An increasing body of literature supports the hypothesis that interferon-beta-mediated changes in the production and activities of the immunoregulatory cytokines interleukin-12 and interleukin-10 are important to the therapeutic benefits of interferon-beta in multiple sclerosis. These data are reviewed here.
Growing evidence indicates that structural cells play a crucial role in the chronic inflammation of autoimmunity by their recruitment of chemokine-dependent cells. Members of the two functional classes of chemokines, inflammatory and homeostatic, seem to be involved in lymphocyte recruitment and survival, and in establishing ectopic lymphoid structures in the target organs of autoimmune diseases. Results from animal models suggest that chemokines are reasonable therapeutic targets in autoimmunity.
Characterising the factors that control the entry of leucocytes into tissue in response to inflammatory or microbial insult continues to generate considerable interest. Of all the tissues studied it is probably that of the CNS which is the most fascinating because of the specialised properties of its blood vessel walls, which constitute the blood-brain barrier (BBB). In health, very few leucocytes penetrate the BBB but in disorders such as MS the barrier becomes compromised with the result that there is an intense infiltration of the CNS by T lymphocytes whose subsequent activity appears to underlie the onset and progression of disease. The purpose of this article is to summarise and assess recent literature pertaining to how lymphocytes bind to cerebral endothelial cells, migrate across the blood vessel walls and enter the CNS parenchyma. Particular emphasis is devoted to the cellular and molecular aspects of these events and addressing the questions of whether certain subsets of circulating T lymphocytes are more favourably disposed than others to CNS infiltration and whether entry is dependent upon the initial expression of distinct groups of adhesion molecules and upon the generation of chemotactic factors. This article also focuses upon identifying the key stages of lymphocyte migration across the BBB and their susceptibility to antagonism by therapeutic agents. It is intended that the review will provide a useful source of information and offer additional insights into the mechanisms controlling lymphocyte passage across the BBB during pathological disturbance.
Multiple sclerosis is an autoimmune demyelinating disease of the human central nervous system with an unknown etiology. Crucial to its pathogenesis is the accumulation and activation of mononuclear cells in the central nervous system. Chemokines and their receptors are proposed to play a major role in the inflammatory recruitment of leukocytes. Besides analyses of relationships between chemokine or chemokine receptor gene polymorphisms and multiple sclerosis susceptibility and severity, analyses of chemokines and their receptors in patients with multiple sclerosis remain descriptive. In clinical material, chemokines and chemokine receptors can be examined in body fluids (blood and cerebrospinal fluid) and in brain tissues obtained via biopsy or autopsy. Research results will be summarized in this review, and a general model of leukocyte migration into the central nervous system under normal and inflammatory conditions will be proposed. Furthermore, opportunities and challenges for future investigations will be identified.
We studied the expression of chemokine receptors CCR1, CCR2, CCR3, CCR5, and CXCR3 on CD4 and CD8 positive T cells, and on CD14 positive monocytes in blood from 10 patients with relapsing-remitting multiple sclerosis (MS) at initiation of interferon (IFN)-beta treatment, after 1 month and after 3 months of treatment. It was found that the expression of CXCR3 on CD4+ and CD8+ T cells was significantly reduced after 3 months of treatment. The expression of other receptors was unaltered. Since CXCR3 cells are enriched in cerebrospinal fluid (CSF), and are detected in lesion material in MS this may represent an important mode of action of interferon-beta in MS.
In this paper we report on the cloning and characterization of mouse CCR8. Like its human homologue, it is predominantly expressed in the thymus. In the periphery, murine CCR8 mRNA was found most abundantly expressed in activated Th2-polarized cells and in NK1.1+CD4+ T cells. Human CCR8 is also preferentially expressed in human Th2-polarized cells and clones. This pattern of expression suggests that CCR8 is part of a Th2-specific gene expression program. The CCR8 ligands I-309 and its mouse homologue T cell activation gene 3 (TCA-3) are potent chemoattractants for Th2-polarized cells. Taken together, these observations strongly suggest that CCR8 plays a role in the control of Th2 responses, and may represent a potential target for treatment of allergic diseases.
Trafficking of inflammatory T cells into the central nervous system (CNS) plays an important role in the pathogenesis of multiple sclerosis, The directional migratory ability of peripheral T cells is associated with interactions of chemokines with their receptors expressed on T cells. In this study, transmigration of peripheral T cells toward a panel of chemokines was examined in patients with multiple sclerosis and healthy individuals using Boyden chemotactic transwells. A significantly increased migratory rate preferentially toward RANTES and MIP-la, but not other chemokines, was found in T cells obtained from multiple sclerosis patients as opposed to healthy individuals (P < 0.001), The migratory T-cell populations represented predominantly Th1/Th0 cells while non-migratory T cells were enriched for Th2-like cells. The study demonstrated further that aberrant migration of multiple sclerosis-derived T cells toward RANTES and MIP-1 alpha resulted from overexpression of their receptors (CCR5) and could be blocked by anti-CCR5 antibodies. These findings have important implications for our understanding of the mechanism underlying aberrant T cell trafficking in multiple sclerosis.
Chemokines and their receptors are important elements for the selective attraction of various subsets of leukocytes. To better understand the selective migration of functional subsets of T cells, chemokine receptor expression was analyzed using monoclonal antibodies, RNase protection assays, and the response to distinct chemokines. Naive T cells expressed only CXC chemokine receptor (CXCR)4, whereas the majority of memory/activated T cells expressed CXCR3, and a small proportion expressed CC chemokine receptor (CCR)3 and CCR5. When polarized T cell lines were analyzed, CXCR3 was found to be expressed at high levels on T helper cell (Th)0s and Th1s and at low levels on Th2s. In contrast, CCR3 and CCR4 were found on Th2s. This was confirmed by functional responses: only Th2s responded with an increase in [Ca2+]i to the CCR3 and CCR4 agonists eotaxin and thymus and activation regulated chemokine (TARC), whereas only Th0s and Th1s responded to low concentrations of the CXCR3 agonists IFN-gamma-inducible protein 10 (IP-10) and monokine induced by IFN-gamma (Mig). Although CCR5 was expressed on both Th1 and Th2 lines, it was absent in several Th2 clones and its expression was markedly influenced by interleukin 2. Chemokine receptor expression and association with Th1 and Th2 phenotypes was affected by other cytokines present during polarization. Transforming growth factor beta inhibited CCR3, but enhanced CCR4 and CCR7 expression, whereas interferon alpha inhibited CCR3 but upregulated CXCR3 and CCR1. These results demonstrate that chemokine receptors are markers of naive and polarized T cell subsets and suggest that flexible programs of chemokine receptor gene expression may control tissue-specific migration of effector T cells.
Several studies have shown that CC chemokines attract T lymphocytes, and that CD45RO+, memory phenotype cells are considered to be the main responders. The results, however, have often been contradictory and the role of lymphocyte activation and proliferation has remained unclear. Using CD45RO+ blood lymphocytes cultured under different stimulatory conditions, we have now studied chemotaxis as well as chemokine receptor expression. Expression of the RANTES/MIP-1 alpha receptor (CC-CKR1) and the MCP-1 receptor (CC-CKR2) was highly correlated with migration toward RANTES, MCP-1, and other CC chemokines, and was strictly dependent on the presence of IL-2 in the culture medium. Migration and receptor expression were rapidly downregulated when IL-2 was withdrawn, but were fully restored when IL-2 was added again. The effect of IL-2 could be partially mimicked by IL-4, IL-10, or IL-12, but not by IL-13, IFN gamma, IL-1 beta, TNF-alpha, or by exposure to anti-CD3, anti-CD28 or phytohemagglutinin. Activation of fully responsive lymphocytes through the TCR/CD3 complex and CD28 antigen actually had the opposite effect. It rapidly downregulated receptor expression and consequent migration even in the presence of IL-2. In contrast to the effects on CC chemokine receptors, stimulation of CD45RO+ T lymphocytes with IL-2 neither induced the expression of the CXC chemokine receptors, IL8-R1 and IL8-R2, nor chemotaxis to IL-8. The prominent role of IL-2 in CC chemokine responsiveness of lymphocytes suggests that IL-2-mediated expansion is a prerequisite for the recruitment of antigen-activated T cells into sites of immune and inflammatory reactions.
T helper cells type 1 (Th1s) that produce interferon-γ predominantly mediate cellular immune responses and are involved in
the development of chronic inflammatory conditions, whereas Th2s which produce large amounts of IL-4 and IL-5 upregulate
IgE production and are prominent in the pathogenesis of allergic diseases. The precise factors determining whether Th1- or
Th2-mediated immune responses preferentially occur at a peripheral site of antigen exposure are largely unknown. Chemokines,
a superfamily of polypeptide mediators, are a key component of the leukocyte recruitment process. Here we report that among
four CXC (CXCR1-4) and five CC (CCR1-5) chemokine receptors analyzed, CXCR3 and CCR5 are preferentially expressed in human
Th1s. In contrast, Th2s preferentially express CCR4 and, to a lesser extent, CCR3. In agreement with the differential chemokine
receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines. The differential expression
of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s. It may also determine
different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors.
Chemokines direct tissue invasion by specific leukocyte populations. Thus, chemokines may play a role in multiple sclerosis (MS), an idiopathic disorder in which the central nervous system (CNS) inflammatory reaction is largely restricted to mononuclear phagocytes and T cells. We asked whether specific chemokines were expressed in the CNS during acute demyelinating events by analyzing cerebrospinal fluid (CSF), whose composition reflects the CNS extracellular space. During MS attacks, we found elevated CSF levels of three chemokines that act toward T cells and mononuclear phagocytes: interferon-gamma-inducible protein of 10 kDa (IP-10); monokine induced by interferon-gamma (Mig); and regulated on activation, normal T-cell expressed and secreted (RANTES). We then investigated whether specific chemokine receptors were expressed by infiltrating cells in demyelinating MS brain lesions and in CSF. CXCR3, an IP-10/Mig receptor, was expressed on lymphocytic cells in virtually every perivascular inflammatory infiltrate in active MS lesions. CCR5, a RANTES receptor, was detected on lymphocytic cells, macrophages, and microglia in actively demyelinating MS brain lesions. Compared with circulating T cells, CSF T cells were significantly enriched for cells expressing CXCR3 or CCR5. Our results imply pathogenic roles for specific chemokine-chemokine receptor interactions in MS and suggest new molecular targets for therapeutic intervention.
Multiple sclerosis (MS) is a T cell-dependent chronic inflammatory disease of the central nervous system. The role of chemokines in MS and its different stages is uncertain. Recent data suggest a bias in expression of chemokine receptors by Th1 vs. Th2 cells; human Th1 clones express CXCR3 and CCR5 and Th2 clones express CCR3 and CCR4. Chemokine receptors expressed by Th1 cells may be important in MS, as increased interferon-gamma (IFN-gamma) precedes clinical attacks, and IFN-gamma injection induces disease exacerbations. We found CXCR3(+) T cells increased in blood of relapsing-remitting MS, and both CCR5(+) and CXCR3(+) T cells increased in progressive MS compared with controls. Furthermore, peripheral blood CCR5(+) T cells secreted high levels of IFN-gamma. In the brain, the CCR5 ligand, MIP-1alpha, was strongly associated with microglia/macrophages, and the CXCR3 ligand, IP-10, was expressed by astrocytes in MS lesions but not unaffected white matter of control or MS subjects. Areas of plaque formation were infiltrated by CCR5-expressing and, to a lesser extent, CXCR3-expressing cells; Interleukin (IL)-18 and IFN-gamma were expressed in demyelinating lesions. No leukocyte expression of CCR3, CCR4, or six other chemokines, or anti-inflammatory cytokines IL-5, IL-10, IL-13, and transforming growth factor-beta was observed. Thus, chemokine receptor expression may be used for immunologic staging of MS and potentially for other chronic autoimmune/inflammatory processes such as rheumatoid arthritis, autoimmune diabetes, or chronic transplant rejection. Furthermore, these results provide a rationale for the use of agents that block CCR5 and/or CXCR3 as a therapeutic approach in the treatment of MS.
T cell responses to the immunodominant peptide (residues 83-99) of myelin basic protein are potentially associated with multiple sclerosis (MS). This study was undertaken to examine whether a common sequence motif(s) exists within the TCR complementarity-determining region (CDR)-3 of T cells recognizing the MBP83-99 peptide. Twenty MBP83-99-reactive T cell clones derived from patients with MS were analyzed for CDR3 sequences, which revealed several shared motifs. Some V beta 13.1 T cell clones derived from different patients with MS were found to contain an identical CDR3 motif, V beta 13.1-LGRAGLTY. Oligonucleotides complementary to the shared CDR3 motifs were used as specific probes to detect identical target CDR3 sequences in a large panel of T cell lines reactive to MBP83-99 and unprimed PBMC. The results revealed that, in contrast to other CDR3 motifs examined, the LGRAGLTY motif was common to T cells recognizing the MBP83-99 peptide, as evident by its expression in the majority of MBP83-99-reactive T cell lines (36/44) and PBMC specimens (15/48) obtained from randomly selected MS patients. The motif was also detected in lower expression in some PBMC specimens from healthy individuals, suggesting the presence of low precursor frequency of T cells expressing this motif in healthy individuals. This study provides new evidence indicating that the identified LGRAGLTY motif is preferentially expressed in MBP83-99-reactive T cells. The findings have important implications in monitoring and targeting MBP83-99-reactive T cells in MS.
Regulated expression of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) plays a role in various physiological processes. To determine in vivo how unbalanced expression of these factors can promote or affect the course of pathologies, we knocked out the mouse gelatinase B gene by replacing the catalytic and zinc-binding domains with an antisense-oriented neomycin resistance gene. Adult gelatinase B-deficient mice and wild-type controls could be induced to develop experimental autoimmune encephalomyelitis (EAE) with similar scores for neurologic disease, blood-brain barrier permeability, and central nervous system histopathology. However, whereas diseased control animals showed necrotizing tail lesions with hyperplasia of osteocartilaginous tissue, adult gelatinase B-deficient mice were resistant to this tail pathology. Gelatinase B-deficient mice younger than 4 weeks of age were significantly less susceptible to the development of EAE than were age matched controls and, even as they aged, they remained resistant to tail lesions. These data illustrate that gelatinase B expression plays a role in the development of the immune system and that, in ontogenesis, the propensity to develop autoimmunity is altered by the absence of this MMP.
Interferon-3-1b (IFN-3-1b, Betaseron) at a dose of 8 million units (8 MU) given subcutaneously (s.c.) every other day to patients with the relapsing-remitting form of multiple sclerosis (MS) lessens the overall frequency of MS attacks by 35%, the frequency of major attacks by 50%, and the number of MS-related hospitalizations by 40% [1, 2]. Benefit persists for at least 5 years, the last assessable time point in the pivotal clinical trial that led to approval of the agent for the treatment of MS [3] by the United States Food and Drug Administration. The agent drastically curtails accumulating disease burden, as assessed by annual magnetic resonance imaging (MRI) scans, again over at least 5 years [4]. Disease activity as measured by serial MRI scans performed at 6-week intervals, is also markedly reduced by IFN-3-lb treatment [4]. IFN-3-1b is currently approved for the treatment of ambulatory MS patients in the United States and Australia; approval in Europe is expected imminently. Trials to determine whether it is effective in progressive MS are underway currently.
Primary infection of macaques with pathogenic isolates of simian immunodeficiency virus (SIV) (as a model of HIV infection in humans) represents a unique opportunity to study early lentivirus/host interactions. We sought to determine whether there is a temporal relationship linking SIV replication and dissemination and the expression of the chemokine RANTES (regulated upon activation normal T cell expressed and secreted) and the SIV/HIV coreceptor CCR5 in different tissues during acute SIV infection of macaques. Four cynomolgus macaques were inoculated intravenously with a pathogenic primary isolate of SIVmac251. RT-PCR was used to monitor the expression of RANTES and CCR5 mRNA in fresh isolated mononuclear cells from blood, lymph node, and bronchoalveolar lavages. These expressions were compared to those of IFN-γ as an indicator of the development of the immune response and to another receptor for RANTES, CCR1, which is not described as a coreceptor for SIV/HIV-1 entry. An enhancement of CCR1/CCR5 mRNA expression was noticed during primary SIVmac251 infection of macaques, mainly in tissue. In the three different compartments investigated, IFN-γ and RANTES overexpression was noticed by the time of systemic viral replication containment. Our results put CCR5 and RANTES mRNA expression back in the context of inflammatory and immune responses to SIV primary infection.
A panel of 90 myelin basic protein (MBP)-specific T-cell lines were derived from peripheral blood of eight patients with multiple sclerosis and four normal subjects. The precursor frequency of MBP-reactive T cells in peripheral blood mononuclear cells ranged from 10(-7) to 9 x 10(-7) (mean, 6.7 x 10(-7)) in the group of patients with multiple sclerosis and from 0.5 x 10(-7) to 9.8 x 10(-7) (mean, 5.6 x 10(-7)) in the control subjects. This difference between the two groups was not statistically significant (p greater than 0.1). These T-cell lines expressed exclusively CD3+CD4+CD8- phenotypes and were restricted predominantly by HLA-DR molecules. When tested with fragments and synthetic peptides of human MBP, these MBP-specific T-cell lines (45 lines for each group) displayed a limited heterogeneous pattern with a biased recognition to peptide 84-102 and the C-terminal peptide 149-171. The reactivity to the 84-102 region of MBP was associated with the HLA-DR2, DRw15 (DRw15,2) haplotype, whereas the recognition to peptide 149-171 did not correlate with a particular HLA-DR allele(s). Furthermore, the majority of T-cell lines (greater than 75%) were found to exhibit substantial cytotoxic activity against MBP-coated target cells, but showing no significant difference between these two groups. This MBP-dependent cytotoxicity was not associated with epitope specificities of the T-cell lines tested.
We have developed an improved method to study the directed migration, or chemotaxis, of rat peripheral blood large granular lymphocytes (LGL) in vitro. A modified Boyden chamber technique was used to measure chemotaxis of LGL through polycarbonate filters that had been coated with different basement membrane components. LGL were found to adhere to collagen types I and IV, laminin and fibronectin. However, only collagen type IV was not in itself chemotactic for LGL. Migrated cells could be identified both morphologically and phenotypically as LGL on collagen type IV-coated filters after incubation with a chemotactic stimulus. LGL were found to display chemotaxis to a number of different stimuli, including the classical chemoattractant agents N-formyl-methionyl-leucyl-phenylalanine, leukotriene B4, and complement fragments present in activated sera. However, the degree of response to these stimuli was much less than that of isolated peripheral blood neutrophils or monocytes. In contrast, all three cell types showed increased chemotaxis to the diacyl glycerol analog 1-oleoyl 2-acetyl glycerol (OAG), which induced a 4-14 fold stimulation of migration. Induction of chemotaxis of LGL by OAG was time and dose-dependent, as confirmed using checkerboard assays. In summary, we have developed a rapid, quantitative method to measure chemotaxis of LGL in vitro. This technique may now be utilized to identify naturally occurring chemoattractants for LGL and to study the intracellular and regulatory events associated with LGL migration.
One method of evaluating the degree of neurologic impairment in MS has been the combination of grades (0 = normal to 5 or 6 = maximal impairment) within 8 Functional Systems (FS) and an overall Disability Status Scale (DSS) that had steps from 0 (normal) to 10 (death due to MS). A new Expanded Disability Status Scale (EDSS) is presented, with each of the former steps (1,2,3 . . . 9) now divided into two (1.0, 1.5, 2.0 . . . 9.5). The lower portion is obligatorily defined by Functional System grades. The FS are Pyramidal, Cerebellar, Brain Stem, Sensory, Bowel & Bladder, Visual, Cerebral, and Other; the Sensory and Bowel & Bladder Systems have been revised. Patterns of FS and relations of FS by type and grade to the DSS are demonstrated.
Gelatinases in inflammatory demyelinating diseases of the central nervous system (CNS) were studied using actively induced experimental autoimmune encephalomyelitis (EAE) in mice as a model system. Clinical disease scores correlated in time and in intensity with pathology parameters such as cytosis in the cerebrospinal fluid (CSF), inflammatory infiltrates, and demyelination in the CNS. Zymographic analysis was employed to measure gelatinases A and B in the CSF from individual animals. According to their apparent molecular weight (MW), gelatinases A and B appeared with a MW of 65 and 95 kDa, respectively. The 65 kDa form was present in all samples, even in those derived from non-induced animals, whereas the 95 kDa form was present only in samples from animals developing EAE. The levels of 95 and 65 kDa gelatinase correlated with the CSF cytosis. In vitro digestion of myelin basic protein (MBP) with gelatinase B and analysis of the cleavage products by protein sequence analysis pinpointed two cleavage sites in conserved regions of MBP. Gelatinase production within the CNS may constitute an important pathogenic mechanism for both the disruption of the blood-brain barrier and the destruction of myelin, as observed in several neuroinflammatory disorders.
The present report compares a variety of T cell purification protocols and chemotaxis procedures in assessing chemokine-induced T cell migration using a microchemotaxis assay. Rapidly purified T cells are capable of directly responding to the beta chemokines macrophage inflammatory protein-1 alpha (MIP-1 alpha), MIP-1 beta, and RANTES in the absence of alpha CD3 stimulation as previously described (Taub, D.D. and Oppenheim, J.J. (1993) Cytokine 5, 175). However, T cell purification schemes involving prolonged 37 degrees C incubations generally produce non-motile T lymphocytes that require stimulation with alpha CD3 antibody for 6-12 h in culture to recover chemotactic mobility. This loss of chemotactic potential appears to be due to prolonged 37 degrees C incubations as rapidly purified T cells lose migratory activity upon incubation at 37 degrees C. Radiolabeled binding analysis revealed that beta chemokine binding sites are downregulated as short as 2 h after incubation at 37 degrees C. T cells require the presence of extracellular matrix molecules to facilitate T cell migration. While many of these proteins permit chemotactic activity, human plasma and foreskin fibronectin were found to be the most effective matrix molecule for T cell migration. Kinetic analysis of T cell activation revealed that 6-12 h of anti-CD3 stimulation was optimal to restore the ability of purified T cells to migrate in response to the chemokines MIP-1 alpha, MIP-1 beta, RANTES, and IL-8. However, rapidly dividing T cells (> or = 48 h post alpha CD3 mAb stimulation) fail to migrate in response to any chemotactic stimulus. Together, these results suggest that the measurement of T cell migration, using microchemotaxis chambers, is a multifactorial process with strict environmental and activation requirements.
Adhesion molecules are important in T-cell trafficking to sites of inflammation. We determined levels of circulating vascular cell adhesion molecule-1 (VCAM-1), L-selectin, and E-selectin in the serum of 147 patients with definite multiple sclerosis of the remitting-relapsing or secondary progressive type. Soluble VCAM-1 and L-selectin concentrations were increased compared to levels in a large group of control subjects. Levels were highest in patients with gadolinium-enhancing lesions on magnetic resonance imaging (VCAM-1: 1,011 +/- 276 vs 626 +/- 87 ng/ml; L-selectin: 1,130 +/- 272 vs 793 +/- 207 ng/ml [mean +/- standard deviation]; p < 0.0001 vs patients without enhancing lesions). Serum levels of soluble tumor necrosis factor receptor (60 kd) were also raised (2.64 +/- 1.23 vs 2.17 +/- 0.69 ng/ml in subjects with other neurological diseases and 2.1 +/- 0.77 ng/ml in healthy control subjects; p < 0.05). Soluble VCAM-1 and L-selectin levels were correlated to concentrations of soluble tumor necrosis factor receptor. In 13 patients with viral encephalitis, similar observations were made. Raised levels of soluble VCAM-1 and L-selectin probably reflect cytokine-induced endothelial cell and T-lymphocyte/monocyte activation occurring in the process of T-cell migration into the central nervous system. Tumor necrosis factor-alpha may be critically involved.
Within the peripheral blood, CD4+CD27- T cells only reside within the CD45RA- (memory or primed) T cell subset. Cells with this phenotype have characteristics of specialized effector T cells according to their cytokine secretion profiles and the expression of tissue-specific adhesion molecules. This subset was previously found to be increased in certain diseases that are associated with immune activation. Therefore we analyzed CD27 expression of peripheral blood and CSF T cells in MS patients. Within the CD4+ T cell subset no differences were seen between MS patients and controls in proportions of CD45RA-CD27- cells. However, when the CD3+ T cell compartment was analyzed, CD27- cells were also found within the CD45RA+ subset. These cells, most likely CD8+, are significantly reduced in PBL and CSF of MS patients as compared with OND patients. In MS and OND groups the level of CD27- cells in peripheral blood correlated significantly with that in CSF, indicating a balanced migration of CD27- cells between the two compartments. In OIND patients, however, this equilibrium was lost. The correlation of the level of CD27+ cells with the amount of intrathecally produced IgG in MS patients may suggest that CD27+ cells are responsible for B cell help in this disease.
The Guillain-Barré syndrome (GBS) and multiple sclerosis (MS) are thought to result from aberrant immune responses to myelin antigens. Recent evidence to implicate the cytokine tumor necrosis factor-alpha (TNF-alpha) and the intercellular adhesion molecule-1 (ICAM-1) in the pathogenesis of these disorders is reviewed. In GBS, elevated serum concentrations of TNF-alpha are detectable in 20 to 50% of patients. TNF-alpha released from autoreactive T cells, macrophages, or microglia may contribute to inflammatory demyelinative processes by upregulating the expression of recognition molecules on antigen-presenting cells; by cytotoxic damage to endothelium; by stimulating the secretion of inflammatory mediators; by directly injuring the myelin sheath; or by interfering with impulse propagation. Its pathogenic potential in GBS is underscored by findings in experimental autoimmune neuritis. Soluble ICAM-1, originating from T cells, macrophages, endothelium, or glial cells, circulates at increased concentrations in serum and cerebrospinal fluid of patients with active MS. ICAM-1 may be crucially involved in the migration of autoreactive T lymphocytes from blood to brain. Whether ICAM-1 can serve as a marker of acute inflammatory events in MS associated with clinical relapses warrants further investigation. TNF-alpha and ICAM-1 could be targets for antigen nonspecific treatment approaches to the inflammatory demyelinating diseases GBS and MS.
The accepted standard treatment of relapsing multiple sclerosis consists of medications for disease symptoms, including treatment for acute exacerbations. However, currently there is no therapy that alters the progression of physical disability associated with this disease. The purpose of this study was to determine whether interferon beta-1a could slow the progressive, irreversible, neurological disability of relapsing multiple sclerosis. Three hundred one patients with relapsing multiple sclerosis were randomized into a double-blinded, placebo-controlled, multicenter phase III trial of interferon beta-1a. Interferon beta-1a, 6.0 million units (30 micrograms¿, was administered by intramuscular injection weekly. The primary outcome variable was time to sustained disability progression of at least 1.0 point on the Kurtzke Expanded Disability Status Scale (EDSS). Interferon beta-1a treatment produced a significant delay in time to sustained EDSS progression (p = 0.02). The Kaplan-Meier estimate of the proportion of patients progressing by the end of 104 weeks was 34.9% in the placebo group and 21.9% in the interferon beta-1a-treated group. Patients treated with interferon beta-1a also had significantly fewer exacerbations (p = 0.03) and a significantly lower number and volume of gadolinium-enhanced brain lesions on magnetic resonance images (p-values ranging between 0.02 and 0.05). Over 2 years, the annual exacerbation rate was 0.90 in placebo-treated patients versus 0.61 in interferon beta-1a-treated patients. There were no major adverse events related to treatment. Interferon beta-1a had a significant beneficial impact in relapsing multiple sclerosis patients by reducing the accumulation of permanent physical disability, exacerbation frequency, and disease activity measured by gadolinium-enhanced lesions on brain magnetic resonance images. This treatment may alter the fundamental course of relapsing multiple sclerosis.
Interferon-beta decreases the relapse rate, relapse severity, progression of neurological disability, and development of new brain lesions observed with brain magnetic resonance imaging in relapsing-remitting multiple sclerosis patients. The mechanism of action of this effect is presently unknown. This study was based on the hypothesis that immunoregulatory effects of interferon-beta may underlie its demonstrated clinical efficacy. The objective of the study was to determine the effect of interferon-beta-1a on the expression of interleukin-10, a cytokine that strongly inhibits cell-mediated immune responses. Interferon-beta-1a induced accumulation of interleukin-10 messenger RNA and protein secretion by cultured peripheral blood mononuclear cells. The observed in vitro effects were similar for healthy control subjects and multiple sclerosis patients. Intramuscular injections of interferon-beta-1a increased serum levels of interleukin-10 at 12 and 24 hours following the injection. Greater increases were induced with 12 x 10(6)-IU than 6 x 10(6)-IU injections. The effect of interferon-beta-1a was relatively specific for interleukin-10, as treatment with interferon-beta-1a did not result in accumulation of transforming growth factor-beta messenger RNA. Upregulation of interleukin-10 represents a possible mechanism of action of interferon-beta's therapeutic effect in relapsing-remitting multiple sclerosis, and has implications for therapy of other autoimmune diseases.
Treatment with interferon beta-1b has substantial clinical benefit in the demyelinating disease multiple sclerosis, yet the mechanism of action in the disease remains largely unknown. Gelatinase A (matrix metalloproteinase-2, 72-kd gelatinase) and B (matrix metalloproteinase-9, 92-kd gelatinase) are matrix metalloproteinases capable of enzymatic digestion of subendothelial basement membrane constituents. In human T cells, interleukin-2 induces gelatinase secretion and enhances gelatinase-dependent migration across an artificial basement membrane-like layer in vitro. Pretreatment of T cells with interferon beta-1b for 48 hours decreased interleukin-2-induced gelatinase production and secretion as determined by zymography. In parallel to the downregulation of gelatinase secretion, pretreatment with interferon beta-1b inhibited T-cell migration across the basement membrane in vitro by up to 90%, but had only a minor impact on cell locomotion per se. For both gelatinase secretion and T-cell migration, the inhibitory effect mediated by exposure to interferon beta-1b was dose dependent. Fluorescence-activated cell sorter analysis also showed that interferon beta-1b downregulates the interleukin-2 receptor alpha-chain and lowered the affinity of interleukin-2 to the cell surface by 30%, which may represent an additional mechanism for the observed effects of interferon beta-1b. The dramatic effects of interferon beta-1b on gelatinase expression and migration raise the possibility that its beneficial effects in multiple sclerosis may result from interference with the capacity of activated T cells to traverse the basement membrane and migrate to the central nervous system.
The chemokine receptors CXCR4 and CCR5 function as coreceptors for HIV-1 entry into CD4+ cells. During the early stages of HIV infection, viral isolates tend to use CCR5 for viral entry, while later isolates tend to use CXCR4. The pattern of expression of these chemokine receptors on T cell subsets and their regulation has important implications for AIDS pathogenesis and lymphocyte recirculation. A mAb to CXCR4, 12G5, showed partial inhibition of chemotaxis and calcium influx induced by SDF-1, the natural ligand of CXCR4. 12G5 stained predominantly the naive, unactivated CD26(low) CD45RA+ CD45R0- T lymphocyte subset of peripheral blood lymphocytes. In contrast, a mAb specific for CCR5, 5C7, stained CD26(high) CD45RA(low) CD45R0+ T lymphocytes, a subset thought to represent previously activated/memory cells. CXCR4 expression was rapidly up-regulated on peripheral blood mononuclear cells during phytohemagglutinin stimulation and interleukin 2 priming, and responsiveness to SDF-1 increased simultaneously. CCR5 expression, however, showed only a gradual increase over 12 days of culture with interleukin 2, while T cell activation with phytohemagglutinin was ineffective. Taken together, the data suggest distinct functions for the two receptors and their ligands in the migration of lymphocyte subsets through lymphoid and nonlymphoid tissues. Furthermore, the largely reciprocal expression of CXCR4 and CCR5 among peripheral blood T cells implies distinct susceptibility of T cell subsets to viral entry by T cell line-tropic versus macrophage-tropic strains during the course of HIV infection.
Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS), characterized by accumulation of mononuclear cells. The pathogenesis of MS is complex and probably involves soluble immune mediators, particularly cytokines, and activated memory T cells, that are thought to migrate into the CNS. During lesion formation in MS, cytokines regulate cell functions, such as cell recruitment and migration. Because the chemokine RANTES play a role in both activating and recruiting leucocytes, particularly memory T cells into inflammatory sites, the authors have assessed RANTES mRNA levels at the site of lesions. Expression levels were analysed in brain samples and compared with neurological, infectious and other controls. RANTES was expressed by activated perivascular memory T cells, predominantly located at the edge of active plaques. While RANTES mRNA was detected in all 17 MS brains analysed, it was only found in six of the 14 control patients and generally at a lower expression level. In view of the regulatory and chemotactic properties of RANTES, these results imply that RANTES in MS lesions may play an important role in the activation and/or selective accumulation of memory T cells and, thereby, in the pathogenic events associated with MS.
Recombinant interferon beta (IFNbeta) benefits patients with relapsing remitting multiple sclerosis (MS), but the mechanisms of action are unknown. We studied in vivo immunologic effects of IFNbeta treatment and their relationship to clinical efficacy. Cytokines were measured in blood and CSF from MS patients participating in a placebo-controlled phase III clinical trial and an open-label phase IV [corrected] tolerability study of IFNbeta-1a. Additionally, immunologic studies were conducted in animals with proteolipid protein (PLP)-induced chronic relapsing experimental autoimmune encephalomyelitis. Single intramuscular (IM) injections of IFNbeta-1a (6 MIU, 30 microg) were associated with significant in vivo upregulation of interleukin-10 (IL-10) and IL-4 but not IFNgamma mRNA in peripheral blood mononuclear cells. Forty-eight hours after each IFNbeta-1a injection, serum IL-10 levels increased and remained elevated for 1 week. IFNbeta-1a recipients in the placebo-controlled phase III clinical trial showed significantly increased concentrations of CSF IL-10 after 2 years of treatment. This response correlated with a favorable therapeutic response. Exposure of PLP-reactive murine T-cell lines to IFNbeta resulted in increased antigen-driven expression of IL-4 and IL-10 and reduced encephalitogenicity. IFNbeta-1a injections induce systemic and intrathecal immunosuppressive cytokines. Myelin-specific T cells treated with IFNbeta-1a demonstrate increased immunosuppressive cytokine expression and reduced encephalitogenicity. The relationship between increased CSF IL-10 and response to therapy suggests that induction of IL-10 is a mechanism underlying IFNbeta-1a effects in MS patients.
Beta-chemokines induce the directional migration of monocytes and T lymphocytes and are thus associated with chronic inflammation. Using immunocytochemistry and in situ hybridisation (ISH) techniques, we have examined the expression of the beta-chemokines monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES (regulated upon activation, normal T cell expressed and secreted) in post-mortem human brain from multiple sclerosis (MS) cases, at different stages of lesion development. In actively demyelinating MS plaques RANTES expression was restricted to the blood vessel endothelium, perivascular cells and surrounding astrocytes, suggesting a role in the recruitment of inflammatory cells from the circulation. MCP-1 was expressed by astrocytes and macrophages within acute MS lesions, but was restricted to reactive astrocytes in the parenchyma surrounding the lesion. MIP-1alpha was expressed by astrocytes and macrophages within the plaque, while MIP-1beta was expressed by macrophages and microglia within the lesion, and by microglia in surrounding white matter. Glial cells may be stimulated to produce chemokines and continue the local inflammatory response by forming chemotactic gradients to attract T cells and mononuclear phagocytes from the circulation and surrounding tissue.
Chemokines play an important role in the selective movement of leucocytes into inflammatory areas and they also activate various cells in inflamed tissues. However, it is unclear which cells are the main sources of chemokines in actual inflammatory diseases, even though both mononuclear cells and non-inflammatory resident cells are able to produce chemokines in vitro and the former cells are also the main target of chemokines. To clarify the roles of chemokines that are produced by mononuclear cells in AD, we measured levels in vivo of mRNA for IL-8 and MIP-1 alpha, as well as the level of regulated upon activation normal T cell expressed and secreted (RANTES) mRNA in freshly isolated peripheral blood mononuclear cells from patients with AD. We compared the results with those from psoriatic patients, and patients without AD who were suffering from other cutaneous diseases and eosinophilia. Levels of mRNAs were determined by semiquantitative reverse transcriptase-polymerase chain reactions. Levels of IL-8 and MIP-1 alpha mRNA were elevated not only in atopic patients but also in non-atopic patients with inflammatory skin disease associated with eosinophilia, compared with levels in psoriatic patients and healthy controls. Levels of RANTES mRNA were similar in atopic patients but they were lower in the other two groups of patients when compared with levels in healthy controls. In atopic patients, the levels of both IL-8 and MIP-1 alpha mRNAs but not of RANTES mRNA decreased with improvements in symptom scores after therapy. These findings suggest that mononuclear cells are not only the target of chemokines but might also play an important role in the pathogenesis of AD by producing IL-8 and MIP-1 alpha.
To define the in vitro effects of interferon beta la (IFN-beta1a) on myelin basic protein (MBP)-reactive T cells and to determine its regulatory mechanism on cytokine networks in patients with MS.
The proliferation and cytokine production of MBP-reactive T-cell clones were measured in thymidine uptake assays and ELISA respectively. The precursor frequency of MBP-reactive T cells was estimated in a microwell culture system.
IFN-beta inhibited the proliferation of established MBP-reactive T-cell clones, which correlated with enhanced production of anti-inflammatory interleukin (IL)-4 and IL-10, and a decrease in tumor necrosis factor alpha (TNF-alpha) and IFN-gamma. When examined with peripheral blood mononuclear cells (PBMCs), IFN-beta was found to reduce the in vitro T-cell responses to MBP, as indicated by the significantly decreased frequency of MBP-reactive T cells. The decreased frequency of MBP-reactive T cells corresponded to an augmented production of IL-4 and IL-10. Although the level of TNF-alpha and IFN-gamma was generally unaltered or decreased, IFN-beta appeared to enhance the production of IFN-gamma in PBMCs derived from some individuals with MS.
Interferon beta la (IFN-beta) suppresses myelin basic protein (MBP)-reactive T cells and induces immune deviation toward the production of T-helper 2 cytokines, which may contribute to its therapeutic benefit in MS. The study also suggests some heterogeneity in MBP-reactive T-cell responses to IFN-beta in different individuals with MS.
Trafficking of inflammatory T cells into the central nervous system (CNS) plays an important role in the pathogenesis of multiple sclerosis. The directional migratory ability of peripheral T cells is associated with interactions of chemokines with their receptors expressed on T cells. In this study, transmigration of peripheral T cells toward a panel of chemokines was examined in patients with multiple sclerosis and healthy individuals using Boyden chemotactic transwells. A significantly increased migratory rate preferentially toward RANTES and MIP-1alpha, but not other chemokines, was found in T cells obtained from multiple sclerosis patients as opposed to healthy individuals (P: < 0.001). The migratory T-cell populations represented predominantly Th1/Th0 cells while non-migratory T cells were enriched for Th2-like cells. The study demonstrated further that aberrant migration of multiple sclerosis-derived T cells toward RANTES and MIP-1 alpha resulted from overexpression of their receptors (CCR5) and could be blocked by anti-CCR5 antibodies. These findings have important implications for our understanding of the mechanism underlying aberrant T cell trafficking in multiple sclerosis.
We performed yearly MRI analyses on 327 of the total 372 patients in a multicenter, randomized, double-blind, placebo-controlled trial of interferon beta-1b (IFNB). Clinical results are presented in the preceding companion paper. Baseline MRI characteristics were the same in all treatment groups. Fifty-two patients at one center formed a cohort for frequent MRIs (one every 6 weeks) for analysis of disease activity. The MRI results support the clinical results in showing a significant reduction in disease activity as measured by numbers of active scans (median 80% reduction, p = 0.0082) and appearance of new lesions. In addition, there was an equally significant reduction in MRI-detected burden of disease in the treatment as compared with placebo groups (mean group difference of 23%, p = 0.001). These results demonstrate that IFNB has made a significant impact on the natural history of MS in these patients.
Mechanisms of action of interferon-b in multiple sclerosis. The finding that CCR5 can be down-regulated in vitro Springer Semin
- B G Arnason
- A Dayal
- Z X Qu
- M A Jensen
- K Genc
- A T Reder
Arnason, B.G., Dayal, A., Qu, Z.X., Jensen, M.A., Genc, K., Reder, A.T., 1996. Mechanisms of action of interferon-b in multiple sclerosis. The finding that CCR5 can be down-regulated in vitro Springer Semin. Immunopathol. 18, 125–148.
over-expression of CCR5 is implicated in the aberrant T CCR51 and CXCR31 T cells are increased in multiple sclerosis and cell trafficking in MS Elevated levels their ligands MIP-and IP-10 are expressed in demyelination brain of both RANTES and MIP-1a were found in cerebrospinal lesions
- K E Balashov
- J B Rottman
- H L Weiner
- W W Hancock
- Zang
and in vivo by IFN-b is particularly interesting because Balashov, K.E., Rottman, J.B., Weiner, H.L., Hancock, W.W., 1999. over-expression of CCR5 is implicated in the aberrant T CCR51 and CXCR31 T cells are increased in multiple sclerosis and cell trafficking in MS (Zang et al., 2000). Elevated levels their ligands MIP-and IP-10 are expressed in demyelination brain of both RANTES and MIP-1a were found in cerebrospinal lesions. Proc. Natl. Acad. Sci. USA 96, 6873–6878.
cells obtained from MS patients as opposed to those Differential expression of chemokine receptors derived from controls. Furthermore, in contrast to other and chemotactic responsiveness of type 1 T helper cells (Th1s) and chemokine receptors, such as CXCR3, that are expressed Th2s
- R Bonecchi
- G Bianchi
- P P Bordignon
- D Ambrosio
- R Lang
- A Borsatti
- S Sozzani
- P Allavena
- P A Gray
- A Mantovani
- F Sinigaglia
both RANTES and MIP-1a, was found in peripheral T Bonecchi, R., Bianchi, G., Bordignon, P.P., D'Ambrosio, D., Lang, R., cells obtained from MS patients as opposed to those Borsatti, A., Sozzani, S., Allavena, P., Gray, P.A., Mantovani, A., Sinigaglia, F., 1998. Differential expression of chemokine receptors derived from controls. Furthermore, in contrast to other and chemotactic responsiveness of type 1 T helper cells (Th1s) and chemokine receptors, such as CXCR3, that are expressed Th2s. J. Exp. Med. 187, 129–134.
van The mechanism of directional T cell migration toward den Opdenakker, concentration gradient of RANTES and MIP-1a is differ-G Resistance of young gelatinase B-deficient mice to experimental autoimmune encephalomyelitis and necrotizing tail
- B Dubois
- S Masure
- U Hurtenbach
- L Paemen
- H Heremans
- J Oord
- R Sciot
- T Meinhardt
- G Hammerling
Dubois, B., Masure, S., Hurtenbach, U., Paemen, L., Heremans, H., van The mechanism of directional T cell migration toward den Oord, J., Sciot, R., Meinhardt, T., Hammerling, G., Opdenakker, concentration gradient of RANTES and MIP-1a is differ-G., Arnold, B., 1999. Resistance of young gelatinase B-deficient mice to experimental autoimmune encephalomyelitis and necrotizing tail Y.C.Q. Zang et al. / Journal of Neuroimmunology 112 (2001) 174 –180
Pullicino, Expression of monocyte chemoattractant protein-1 and other b Intramuscular inter-chemokines by resident glia and inflammatory cells in multiple feron b-1a for disease progression in relapsing multiple sclerosis. sclerosis lesions
- R P Kinkel
- M K Mass
- F E Munschauer
- R L Priore
- .-P M Scherokman
- B J Whitham
Kinkel, R.P., Mass, M.K., Munschauer, F.E., Priore, R.L., Pullicino, Expression of monocyte chemoattractant protein-1 and other b-P.M., Scherokman, B.J., Whitham, R.H., 1996. Intramuscular inter-chemokines by resident glia and inflammatory cells in multiple feron b-1a for disease progression in relapsing multiple sclerosis. sclerosis lesions. J. Neuroimmunol. 84, 238–249.
Chemotaxis of T lymphocytes on extracellular matrix proteins.
- Taub D.
- Key M.L.
- Clark D.
- Turcovski-Corrales S.M.
an Expanded Disability Status Score (EDSS). Neurology 33, 1444– Taub, D., Key, M.L., Clark, D., Turcovski-Corrales, S.M., 1995. 1452. Chemotaxis of T lymphocytes on extracellular matrix proteins. J. Leppert, D., Waubant, E., Burk, M.R., Oksenberg, J.R., Hauser, S.L., Immunol. Meth. 184, 187–198.
Aberrant T Cell Migration toward MIP-1a in Patients with Multiple Sclerosis: Over-Expression of 1990. An improved in vitro assay to quantitate chemotaxis of rat Chemokine Receptor CCR5
- J Zhang
- Rantes
- A Pilaro
- T J Sayers
- K L Mccormick
- C W Reynold
- R H Wilrout
Zhang, J., 2000. Aberrant T Cell Migration toward RANTES and Pilaro, A., Sayers, T.J., McCormick, K.L., Reynold, C.W., Wilrout, R.H., MIP-1a in Patients with Multiple Sclerosis: Over-Expression of 1990. An improved in vitro assay to quantitate chemotaxis of rat Chemokine Receptor CCR5. Brain 123, 1874–1882.
Myelin basic protein-specific T lymphocytes in Poser, multiple sclerosis and controls: precursor frequency, fine specificity
- J C M Raus
- D W Paty
- L Scheinberg
- W I Mcdonald
- F A Davis
Raus, J., 1992. Myelin basic protein-specific T lymphocytes in Poser, C.M., Paty, D.W., Scheinberg, L., McDonald, W.I., Davis, F.A., multiple sclerosis and controls: precursor frequency, fine specificity, Ebers, G.C., Johnson, K.P., Sibley, W.A., Silberberg, D.H., Tourtellot-and cytotoxicity. Ann. Neurol. 32, 330–338.
New diagnostic criteria for multiple sclerosis: Guide-Zingoni, A lines for research protocols
- W W Te
- H Soto
- J A Hedrick
- A Stoppacciaro
- C T Storlazzi
te, W.W., 1983. New diagnostic criteria for multiple sclerosis: Guide-Zingoni, A., Soto, H., Hedrick, J.A., Stoppacciaro, A., Storlazzi, C.T., lines for research protocols. Ann. Neurol. 13, 227–236.
Interferon-b induces interleukin-10 receptor CCR8 is preferentially expressed in Th2 but not in Th1 cells. expression: Relevance to multiple sclerosis
- F Sinigaglia
- D Ambrosio
- O Garra
- A Robinson
- D Rocchi
- R A Rudick
- R M Ransohoff
- R Peppler
- S Medendorp
- M Santoni
- A Zlotnik
- A Napolitano
Sinigaglia, F., D'Ambrosio, D., O'Garra, A., Robinson, D., Rocchi, Rudick, R.A., Ransohoff, R.M., Peppler, R., Vanderbrug Medendorp, S., M., Santoni, A., Zlotnik, A., Napolitano, M., 1998. The chemokine Lehmann, P., Alam, J.J., 1996. Interferon-b induces interleukin-10 receptor CCR8 is preferentially expressed in Th2 but not in Th1 cells. expression: Relevance to multiple sclerosis. Ann. Neurol. 40, 618– J.
An improved in vitro assay to quantitate chemotaxis of rat peripheral blood large granular lymphocytes (LGL).
- Pilaro A.
- Sayers T.J.
- McCormick K.L.
- Reynold C.W.
- Wilrout R.H.
Interferon β-1b is effective in relapsing–remitting multiple sclerosis. I. Clinical results of a multicenter, randomized, double blind, placebo-controlled trial.
Chemotaxis of T lymphocytes on extracellular matrix proteins
- Taub