IL-8 Secretion In Cells Research Articles

Opioid growth factor and low-dose naltrexone impair central nervous system infiltration by CD4 + T lymphocytes in established experimental autoimmune encephalomyelitis, a model of multiple sclerosis.

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS), characterized by infiltrating myelin-reactive T lymphocytes and demyelinating lesions. Experimental autoimmune encephalomyelitis (EAE) is the animal model widely utilized to study MS. EAE is mediated by CD4(+) T cells and can be induced in EAE-susceptible mice through immunization with a myelin antigen, such as proteolipid protein 139-151 (PLP139-151) in SJL mice. In this PLP-induced EAE model, autoreactive CD4(+) T cells migrate from peripheral tissues into the CNS where they are reactivated resulting in CNS damage. Th1 and Th17 cells produce the pro-inflammatory cytokines IFNγ and IL-17, respectively, that have been shown to have pathogenic roles in EAE and MS. Anti-inflammatory Th2, IL-4 secreting cells, have been indicated to inhibit EAE exacerbation. However, given the inflammatory environment of EAE, Th2 effector cells are outnumbered by Th1/Th17 cells. Regulatory CD4(+) T cells suppress immune reactions and have been demonstrated to be dysfunctional in MS patients. Opioid growth factor (OGF), chemically termed [Met(5)]-enkephalin, is a negative growth factor that interacts with the OGF receptor. The OGF-OGFr axis can be activated through exogenous administration of OGF or a low dosage of naltrexone (LDN), an opioid antagonist. We have previously demonstrated that modulation of the OGF-OGFr axis results in alleviation from relapse-remitting EAE, and that CNS-infiltrating CD3(+) T cells are diminished with exogenous OGF or intermittent blockade with LDN administration. In this paper, we aimed to determine whether OGF or LDN alter the Th effector responses of CD4(+) T lymphocytes within the CNS in established EAE. We report in these studies that the numbers of CD4(+) T lymphocytes in the CNS of EAE mice are decreased following treatment with OGF for five days but not LDN. However, modulation of the OGF-OGFr axis did not result in changes to CD4(+) Th effector cell responses in the CNS of EAE mice.

Influence of 1 Alpha, 25-Dihydroxyvitamin D3 on T Helper 17 Cells and Related Cytokines in Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease. Emerging data suggests that T helper 17 (Th17) cells play a pathogenic role in SLE and the increased number of these cells correlates with disease activity. In recent years, 1α, 25-dihydroxyvitamin D3 (1,25VitD3) has been considered as an immunomodulatory factor. To investigate the effect of 1,25VitD3 on Th17 cells and on the expression of related cytokines in SLE patients. Thirty SLE patients (newly diagnosed or in remission) were sampled for 10 ml whole blood to isolate peripheral blood mononuclear cells (PBMCs) using Ficoll-Hypaque density gradient centrifugation. Isolated cells were cultured in the presence and absence of 50 nM 1,25VitD3. After incubation, cells were harvested and stimulated for 4-5 hours with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A. IL-17 secreting cells were analyzed by flowcytometry. RNA was extracted from cultured cells, cDNA was synthesized, and the expression levels of IL-6, IL-17, IL-23 and TGF-β genes were assessed by real-time PCR. The percentage of Th17 cells (CD3+CD8- IL-17+ T cells) decreased significantly in 1,25VitD3-treated cells (3.67 ± 2.43%) compared to untreated cells (4.65 ± 2.75%)( p=0.003). The expression of TGF-β up regulated (1.38-fold) and the expression of IL-6 (50%), IL-17 (27%) and IL-23 (64%) down regulated after 1,25VitD3 treatment. This study showed that 1,25VitD3 modulates Th17 related pathways in SLE patients and revealed the immunomodulatory effect of 1,25VitD3 on the Th17 mediated autoimmunity.

Role of interleukin-1 receptor antagonist in regulating specific Hepatitis B antibody production (IRM12P.646)

Abstract While exogenous interleukin-1 beta was reported to facilitate antibody production, the function and mechanism of its conventional suppressor interleukin-1 receptor antagonist (IL-1Ra) in regulating specific antibody remain largely unknow. We showed that, upon aluminium-conjugated commercial hepatitis B surface antigen vaccination, hepatitis B antibody response was considerably amplified in IL-1Ra-/- mice, demonstrated by higher HBsAb level, increased germinal center and HBsAb secreting B cell numbers in dLN but not spleen. Mechanismly, IL-1 signalling pathway was, unexpectedly, found not to be involved in the process, evident by equivalent HBsAb titre in nalp3-/-, caspase-1-/- and IL-1R-/- mice as in wt. Surprisingly, the IL-17 production was amplified in IL-1Ra-/- mice, demonstrated by higher serum IL-17 level and higher number and frequency of IL-17 secreting cells in dLN of IL-1Ra-/- mice. Moreover, HBsAb level in IL1rn-/- x IL17-/- mice was comparable to that in wt mice, suggesting the essential role of IL-17 in regulating the elevated HBsAb level in IL-1Ra-/- mice. Furthermore, CD4 T cells was identified as the main source of IL-17 production in dLN of IL-1Ra-/- mice, indicating the essential role of IL-17 secreting CD4 T cells in upregulating HbsAb production. Therefore, the increased HBsAb production in IL-1Ra-/- was irrelevant to IL-1 pathway but dependend on evelated IL-17. We have thus proposed a novel mechanism of IL-1Ra in regulating specific antibody production.

Jak/STAT signaling inhibition enhances costimulation blockade and promotes transplant acceptance (TRAN1P.943)

Abstract Transplant tolerance induction through costimulation blockade (CoB) remains an elusive goal. Recent evidence suggests that inflammatory cytokines (IC) contribute to the activation of alloreactive T cells in a CD28- and CD40-independent manner. We aimed to delineate the possible synergism between CD28 blockade and inhibition of IC signaling via Jak/STAT inhibition. First, we measured the impact on mouse T cell activation via quantification of IL-2 secreting cells at 24h post in vitro stimulation. Although CTLA4-Ig diminished the proportion of IL-2+ cells, the concomitant addition of the supernatant (MATSup) from maturing dendritic cells completely abolished this effect. This suggested an early impact of IC on T cell activation. MATSup counteracted the anti-proliferative effect of CTLA4-Ig on both CD4 and CD8 T cells. However, addition of the Jak3/1 inhibitor Tofacitinib (Tofa) completely restored the effect of CTLA4-Ig in presence of MATSup. In a BALB/c to B6 heart transplant model a short course of Tofa (d0-6) synergized with CTLA4-Ig promoting long term graft survival (MST untreated: 11d, CTLA4-Ig only: 36d, Tofa+CTLA4: 114d). Survival was associated to lower Th1 cell production and, unexpectedly, an increment in graft infiltrating Treg. Overall, our results indicate that inflammatory cytokines counteracts the efficacy of CoB. However, this effect can be neutralized via transient inhibition of Jak signaling - a promising new immunoregulatory strategy we define Enhanced CoB.

Sensitizing B- and T- cell Lymphoma Cells to Paclitaxel/Abraxane-Induced Death by AS101 via Inhibition of the VLA-4-IL10-Survivin Axis.

Cancer cell resistance to chemotherapy is a major concern in clinical oncology, resulting in increased tumor growth and decreased patient survival. Manipulation of apoptosis has emerged as a new therapeutic strategy to eliminate cancer cells. The focus of this study resides within a novel approach to target survivin, an integrator of both cell death and mitosis. This protein plays a pivotal role in the resistance of tumors to chemotherapy, especially to paclitaxel. The data herein demonstrate an indirect repression of survivin in both B- and T-cell lymphoma and human NHL by the nontoxic tellurium compound, AS101 [ammonium trichloro(dioxoethylene-o,o')tellurate], via inhibition of tumor autocrine IL10-STAT3-Survivin signaling. As a result of survivin abrogation, sensitization of lymphomas to paclitaxel or to Abraxane, the new albumin-stabilized nanoparticle formulation of paclitaxel, occurs both in vitro and in vivo. Importantly, inhibition of lymphoma cell IL10 secretion is mediated by inactivation of the VLA-4 integrin, recently shown to be an important target of AS101. This activity is followed by inhibition of the PI3K-AKT axis that mediates IL10 suppression. Because a wide variety of lymphomas and other tumor types express VLA-4 and secrete IL10 in an autocrine manner, inhibition of survivin with a small nontoxic agent has vast clinical significance in modulating chemosensitivity in many tumor types. Combination therapy with AS101 and paclitaxel has novel therapeutic potential targeting deregulated active pathways in lymphoma, overcoming endogenous resistance to apoptosis.

Th17 plasticity and its changes associated with inflammatory bowel disease.

CD4 T helper (Th) cell differentiation into distinct T cell subsets is critical to the normal function of the immune system. Until recently, the paradigm held that naïve T cells differentiated into distinct subsets under the guidance of environmental cues (e.g., cytokines) and that once polarized, these cells were committed to a particular functional state. However, the existence of transdifferentiated T cell populations, which express signature transcription factors and cytokines associated with more than one Th subset, challenges the immutability of T helper subsets and suggests that plasticity is a feature of multifaceted immune responses. How this process impacts immune dysregulation in diseases such as inflammatory bowel diseases (IBD) and the machinery that underlies this process is far from fully understood. Interleukin (IL)-17 secreting helper T (Th17) cells have been heavily implicated in tissue-specific immune pathology including murine models of IBD, human Crohn's disease and ulcerative colitis. Plasticity within this subset is suggested by the existence of IL-17 secreting cells, which, can also secrete interferon-γ, the signature cytokine for Th1 cells or, can co-express the anti-inflammatory transcription factor forkhead box p3, a signature transcription factor of regulatory T cells. In this review we mainly discuss evidence for Th17 plasticity, mechanisms, which govern it, and highlight the potential to therapeutically target this process in human IBD.

Abstract 4163: Chemokine and cytokine levels among lung cancer cases responsive to HPV antigen stimulation

Abstract Purpose: Human papillomavirus (HPV), the causative agent for anogenital and certain head and neck cancers, has been detected in lung tumors, but its role as an etiologic agent for lung cancer remains controversial. The lung is often subjected to exogenous inflammatory insults, and patients with chronic lung inflammation have an increased risk for cancer. In addition to the genomic instability that can be caused by HPV oncoproteins, the immunomodulatory impacts of HPV infection may foster a pro-inflammatory state that increases lung cancer risk. We sought to identify such immunomodulatory effects by examining the differences in lymphocyte and cytokine/chemokine profiles between lung cancer cases and HPV-vaccinated healthy controls. Methods: Peripheral blood mononuclear cells were isolated from 22 newly diagnosed lung cancer cases and 11 healthy controls who had prior exposure to HPV antigens (immunized with Gardasil). Cells were enumerated via a hemacytometer, and were exposed to media alone or to 3 doses of Gardasil for 20 hours as an HPV challenge. Interleukin (IL-) 4, IFN-γ, and IL-17A secreting cells were then quantified using enzyme linked immunocell spot (ELISpot) analysis. Individuals were categorized as HPV-responsive if the HPV challenge resulted in an increase in at least 2 types of secreting cells, compared to media-exposed cells. The Milliplex human cytokine detection kit was used to assess circulating levels of 38 key cytokines in the plasma of cases and controls. Cytokine levels among HPV-responsive and HPV-non-responsive cases were compared. Survival analysis was also conducted. Results: All 11 HPV-vaccinated controls demonstrated an increase in cells secreting IFN-γ and IL-4. Of the 22 cases, 13 (59%) were classified as HPV-responsive, with 2 cases displaying an increase in IFN-γ and IL-4 secreting cells upon HPV challenge. Levels of TGF-α, IL-10, IL-3, IL-5, and IL-7 were significantly higher among HPV-responsive compared to non-responsive cases. Significant survival differences were also observed by HPV-responsiveness and cytokine levels. Conclusions: HPV clearly has substantial effects on the host's systemic immune response. Our study may explain whether and how the pathogenic process differs between HPV-infected and -uninfected lung tumors. Citation Format: Michael E. Scheurer, E Susan Amirian, Paul Porter, David B. Corry. Chemokine and cytokine levels among lung cancer cases responsive to HPV antigen stimulation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4163. doi:10.1158/1538-7445.AM2014-4163

Porphyromonas gingivalis stimulates IL-6 and IL-8 secretion in GMSM-K, HSC-3 and H413 oral epithelial cells

Infection of oral epithelial cells with periodontopathogenic bacteria results in the production of pro-inflammatory cytokines involved in the initiation and progression of periodontal disease. The purpose of this study was to examine the release of interleukin (IL)-6 and IL-8 by oral epithelial cells after exposure to Porphyromonas gingivalis. Non-tumor-derived, immortalized human GMSM-K cells, and human oral squamous cell carcinoma, HSC-3 and H413 cells, were co-cultured with live and heat-inactivated P. gingivalis 2561 (ATCC 33277) and W83 (ATCC BAA-308™). IL-6 and IL-8 were quantified in the culture supernatants after 6 and 24 h. The basal levels of both cytokines and the responses to P. gingivalis were strongly dependent on cell type. GMSM-K cells produced less IL-8 than HSC-3 and H413 cells. Live P. gingivalis induced significant IL-6 and IL-8 secretion in GMSM-K and HSC-3 cells, and heat-inactivation of bacteria enhanced greatly IL-6 and IL-8 stimulation in these cells. Uninfected H413 cells produced high levels of IL-6 and IL-8, but were not responsive to live P. gingivalis; heat-inactivated P. gingivalis up-regulated IL-6 and IL-8 secretion in these cells. Since base-line secretion of IL-6 and IL-8, and responses to P. gingivalis depend on the cell type, conclusions on the responses to P. gingivalis should not be based on studies with a single cell type.

AB0024 Regulatory B Cell Functions Are Altered in Patients with Rheumatoid Arthritis and the Impairment of B Cell IL-10 Secretion is not Corrected by Efficient Therapies

BackgroundWe previously showed that regulatory IL-10 producing B cells (B10) were decreased in patients with rheumatoid arthritis (RA) and that B10 cells were inversely correlated with disease activity (DAS28) and...

1036-LBO: BATF plays a crucial role in induction of autoimmunity and BOS development following administration of antibodies to MHC

Development of antibodies to donor specific antigen following transplantation is an increased risk factor for development of chronic rejection. We have previously demonstrated that administration of anti-MHC antibodies (ABs) results in the induction of immune responses against self antigens (K α 1 tubulin (KAT) and collagen V (Col V) leading to development of OAD mediated by IL-17. The overall objective of the study was to define the molecular mechanisms that contribute to OAD development . Basic leucine zipper transcription factor, ATF-like (BATF) is an upstream signaling mediator that controls development of TH17 responses and induction of autoimmunity. BATF−/− mice and wild type mice were administered anti-MHC ABs endobronchially and analyzed for the development of OAD on day 30. Humoral and cellular immune responses against the self antigens (KAT and Col V) were analyzed by ELISA and ELISPOT respectively. B cell infiltration in the lungs were analyzed by flow cytometry and IL-6expression was analyze by qRT-PCR. Histopathological analysis of lungs from BATF−/− demonstrated no significant development of OAD following administration anti-MHC ABs. BATF−/− resulted in a eight fold reduction in cellular infiltration ( p < 0.05), 12 fold reduction in epithelial metaplasia and 10.4 fold reduction in fibrosis compared to controls. We observed a 93% reduction in the B cell infiltration and 3.8 fold reduction in the expression of IL-6 in the BATF deficient animals compared to controls. Moreover we observed a complete abrogation of humoral immune responses against the self antigen and 4.2 fold reduction in the IL-17 secreting cells against the self antigens in BATF−/− mice. Based on these results, we conclude that BATF play an important role in induction of immune responses to self antigens and immunopathogenesis of OAD following administration of anti-MHC. Therefore, strategies to block BATF signaling should be considered for preventing the development of chronic rejection.

Identification of selective small molecule inhibitors of the nucleotide-binding oligomerization domain 1 (NOD1) signaling pathway.

NOD1 is an intracellular pattern recognition receptor that recognizes diaminopimelic acid (DAP), a peptidoglycan component in gram negative bacteria. Upon ligand binding, NOD1 assembles with receptor-interacting protein (RIP)-2 kinase and initiates a signaling cascade leading to the production of pro-inflammatory cytokines. Increased NOD1 signaling has been associated with a variety of inflammatory disorders suggesting that small-molecule inhibitors of this signaling complex may have therapeutic utility. We utilized a cell-based screening approach with extensive selectivity profiling to search for small molecule inhibitors of the NOD1 signaling pathway. Via this process we identified three distinct chemical series, xanthines (SB711), quinazolininones (GSK223) and aminobenzothiazoles (GSK966) that selectively inhibited iE-DAP-stimulated IL-8 release via the NOD1 signaling pathway. All three of the newly identified compound series failed to block IL-8 secretion in cells following stimulation with ligands for TNF receptor, TLR2 or NOD2 and, in addition, none of the compound series directly inhibited RIP2 kinase activity. Our initial exploration of the structure-activity relationship and physicochemical properties of the three series directed our focus to the quinazolininone biarylsulfonamides (GSK223). Further investigation allowed for the identification of significantly more potent analogs with the largest boost in activity achieved by fluoro to chloro replacement on the central aryl ring. These results indicate that the NOD1 signaling pathway, similarly to activation of NOD2, is amenable to modulation by small molecules that do not target RIP2 kinase. These compounds should prove useful tools to investigate the importance of NOD1 activation in various inflammatory processes and have potential clinical utility in diseases driven by hyperactive NOD1 signaling.

Sa1773 Caspase-11 Enhances Killing of Colitogenic Escherichia coli by Macrophages, but Does Not Affect Development of Experimental Colitis

WT Balb/C mice (p<0.001 for each cytokine btw uninfected control and helminth-infected; N=at least 3 independent experiments), while no regulation of IFNγ and modest induction of IL4 or IL10 was seen in STAT6-/mice. Helminths did not induce TGFβ production in STAT6-/animals. In an acute GVHD and colitis model, helminths regulated WT donor T cell Th1 cytokine generation, serum Th1 cytokines, colitis and survival in WT and not STAT6-/recipients (p<0.05 for each parameter btw. helminth infected WT and helminthinfected STAT6-/recipients, multiple experiments). Helminth infection was associated with the induction of donor and recipient FoxP3+ regulatory T cells (Tregs) in WT and not STAT6-/bone marrow transplanted mice (p<0.05 btw. helminth infected WT and helminthinfected STAT6-/recipients, multiple experiments). Addition of TGFβ to STAT6-/T cell cultures restored T cell IL10 production and Treg generation that regulate Th1 inflammation. Conclusions: STAT6 is a critical transcription factor directing mucosal TGFβ producing Th3 cell generation. Helminths utilize STAT6 pathway to induce mucosal Tregs, trigger mucosal T cell IL10 secretion and regulate alloreactive colitis in a TGFβ dependent manner.

Sa1778 Constitutive Type I Interferon, via STAT1 Activation, Selectively Promotes Regulatory T Cell Function in the Healthy Human Intestine, but Not in Inflammatory Bowel Disease

WT Balb/C mice (p<0.001 for each cytokine btw uninfected control and helminth-infected; N=at least 3 independent experiments), while no regulation of IFNγ and modest induction of IL4 or IL10 was seen in STAT6-/mice. Helminths did not induce TGFβ production in STAT6-/animals. In an acute GVHD and colitis model, helminths regulated WT donor T cell Th1 cytokine generation, serum Th1 cytokines, colitis and survival in WT and not STAT6-/recipients (p<0.05 for each parameter btw. helminth infected WT and helminthinfected STAT6-/recipients, multiple experiments). Helminth infection was associated with the induction of donor and recipient FoxP3+ regulatory T cells (Tregs) in WT and not STAT6-/bone marrow transplanted mice (p<0.05 btw. helminth infected WT and helminthinfected STAT6-/recipients, multiple experiments). Addition of TGFβ to STAT6-/T cell cultures restored T cell IL10 production and Treg generation that regulate Th1 inflammation. Conclusions: STAT6 is a critical transcription factor directing mucosal TGFβ producing Th3 cell generation. Helminths utilize STAT6 pathway to induce mucosal Tregs, trigger mucosal T cell IL10 secretion and regulate alloreactive colitis in a TGFβ dependent manner.

Bone marrow-derived mesenchymal stem cells affect immunologic profiling of interleukin-17-secreting cells in a chemical burn mouse model.

PurposeThis study investigated interleukin (IL)-17-secreting cell involvement in sterile inflammation, and evaluated the effect of mesenchymal stem cells (MSCs) on IL-17-secreting cell immunologic profiling.MethodsTwenty mice were sacrificed at time points of 6 hours, 1 day, 1 week, and 3 weeks (each group, n = 5) after the cornea was chemically injured with 0.5N NaOH; IL-17 changes in the cornea were evaluated using enzyme-linked immunosorbent assay. Further, IL-17 secreting cells were assessed in the cervical lymph nodes by a flow cytometer. Rat MSCs were applied intraperitoneally in a burn model (n = 10), IL-17-secreting T helper 17 (Th17) cell and non-Th17 cell changes were checked using a flow cytometer in both cornea and cervical lymph nodes at 1week, and compared with those in the positive control (n = 10).ResultsIL-17 was highest in the cornea at 1 week, while, in the cervical lymph nodes, IL-17-secreting cells showed early increase at 6 hours, and maintained the increase through 1 day to 1 week, and levels returned to the basal level at 3 weeks. Specifically, the non-Th17 cells secreted IL-17 earlier than the Th17 cells. When the MSCs were applied, IL-17 secretion was reduced in CD3(+)CD4(-)CD8(-), CD3(+)CD4(+)CD8(-), and CD3(+) CD4(-)CD8(+) cells of the cervical lymph nodes by 53.7%, 43.8%, and 50.8%, respectively. However, in the cornea, IL-17 secretion of CD3(+)CD4(-)CD8(-) cells was completely blocked.ConclusionsThe results indicated that both IL-17-secreting non-Th17 and Th17 cells were involved in the chemical burn model, and MSCs appeared to mainly modulate non-Th17 cells and also partially suppress the Th17 cells.

PReS-FINAL-2069: T cells secreting granulocyte-macrophage colony stimulating factor (GM-CSF) within the inflammed joint originate from an "EX-Th17" population

Since 2003, the established paradigm of T cell immunology has defined interleukin (IL)-17 as a dominant Th17 cell derived cytokine driving autoimmune disease. Recent murine studies have challenged this, identifying GM-CSF as a Th17 related cytokine necessary and sufficient for the induction of autoimmunity. The origin of GM-CSF+ T cells and their relationship with IL-17 secreting cells is unclear in human autoimmune disease. Trials of biologic agents targeting the GM-CSF pathway show promise in rheumatoid arthritis, so it is important to establish if GM-CSF contributes to the inflammatory environment of the arthritic joint in Juvenile Idiopathic Arthritis (JIA).

Characteristics of the cellular immune response in HIV/HCV patients with hemophilia during peginterferon/ribavirin therapy in southern China

ObjectiveThe objectives of the study are to characterize the cellular immune response in hepatitis C virus (HCV) genotype 1 and HIV co-infected patients with hemophilia in southern China during treatment with interferon and ribavirin and to study its correlation with the virologic response (VR). Thirty-six HCV genotype 1 and HIV co-infected patients with hemophilia in southern China were enrolled into the study. Using an ELISpot assay, HCV antigen-specific interferon (IFN) γ, interleukin (IL) 2, IL-4, and IL-10 secreting cells were measured in peripheral blood mononuclear cells. Single nucleotide polymorphisms of IL28B were determined, and immunological, virologic, and clinical variables were collected to identify factors associated with HCV-sustained VR (SVR) at week 72 after treatment. At baseline, there were no significant differences in IFN-γ and IL-2 mediated immune responses in subjects with VR versus non-responders. Higher IL-10 specific responses to NS3 were observed in VR patients. Subjects who had significant decreases in IL-10 responses at week 72 compared with baseline for NS3 and NS5 were more likely to be VR. In SVR, IL-2 production decreased moderately, and the levels of IL-4 were low throughout. The main correlation for SVR in genotype-l infected subjects was sustained HCV-specific IFN-γ responses through the whole 72-week period. In subjects with HIV and HCV co-infection combined with hemophilia, IL28B genotype CC, a decrease in HCV specific IL-l0 and IL-2 responses, and the maintenance of IFN-γ responses during treatment were associated with a 12- or 72-week VR.

Nanoparticle Vaccines Encompassing the Respiratory Syncytial Virus (RSV) G Protein CX3C Chemokine Motif Induce Robust Immunity Protecting from Challenge and Disease

Nanoparticle vaccines were produced using layer-by-layer fabrication and incorporating respiratory syncytial virus (RSV) G protein polypeptides comprising the CX3C chemokine motif. BALB/c mice immunized with G protein nanoparticle vaccines produced a neutralizing antibody response that inhibited RSV replication in the lungs following RSV challenge. ELISPOT analysis showed that G nanoparticle vaccinated mice had increased levels of RSV G protein-specific IL-4 and IFN-γ secreting cells compared to controls following RSV challenge. Remarkably, RSV challenge of G protein nanoparticle vaccinated mice resulted in increased RSV M2-specific IL-4 and IFN-γ secreting T cells, and increased M2-specific H-2Kd-tetramer positive CD8+ T cells in the lungs compared to controls. Cell type analysis showed vaccination was not associated with increased pulmonary eosinophilia following RSV challenge. These results demonstrate that vaccination of mice with the RSV G protein nanoparticle vaccines induces a potent neutralizing antibody response, increased G protein- and M2- specific T cell responses, and a reduction in RSV disease pathogenesis.

107 : Development of vaccination strategies for Yersinia Pestis that triggers T cell immune response

Introduction: Yersinia pestis , a Gram negative bacterium, is the etiological agent of pneumonic and bubonic plague. The current efforts are aimed at evaluating whether our vaccine protocol primes memory T cells that rapidly produce IFN- γ and/or perforin in response to peptide or conjugate stimulation. Method: In the present study, IFN- γ and IL-4 production was measured by ELISpot assay, parallel with the ELISA based measurements of total cytokine production in splenocytes, Lamina propria and peyer’s patches of mice, immunized by different constructs of B and T cell epitopes of F1 and V antigens entrapped in PLGA microspheres intranasally. The ratio of CD4+/CD8+ cell population expressing IFN- γ and perforin was also measured by intracellular staining. Result: We observed that three B–T constructs of F1 and seven B–T constructs of V antigens produced a high number of IFN- γ secreting cells as compared to native antigen and a low number of IL-4 secreting cells. B–T conjugates of F1 and V antigens showed significantly high ( p + IFN- γ + cells as compared to CD8 + IFN- γ + cells. Conclusion: Our study demonstrated that IFN- γ is an effector molecule of activated CD4 + and CD8 + T cells arisen from B–T constructs of F1 and V antigens for the antibacterial activity as an improved plague vaccine.

THU0294 Regulatory T cells/TH17 balance in the pathogenesis of pediatric behçet disease

BackgroundBehçet disease (BD) is an idiopathic systemic inflammatory disorder of unknown origin characterizedby recurrent attacks involving the muco-cutaneous, ocular, intestinal, vascular and nervous systems. In adult, it has been shown...

Fingolimod Modulates Peripheral Effector and Regulatory T Cells in MS Patients

Multiple sclerosis (MS) is a complex neurological disease where, in genetically predisposed individuals, the unbalanced interplay between pathogenic and regulatory T cells will result in the progression of the autoimmune assault to neural antigens. Fingolimod (FTY720), an oral sphingosine 1-phosphate modulator recently approved for the treatment of MS, inhibits the egress of T cells from lymph nodes acting specifically on naïve and memory T cells and sparing effector T cells. Here we characterized IL-17 and IFNγ producing effector CD4 and CD8 positive T cells as well as CD4 positive CD25highCD127low regulatory T cells in MS patients before and 1 month after treatment was started. We observed that fingolimod did not significantly affect the percentage of CCR6 and CD161 positive T cells in both CD4 and CD8 compartments. In contrast, it significantly reduced the levels of both CD4+ CCR6+ CD161+ and CD8+ CCR6+ CD161+ producing IFNγ alone or in combination with IL-17. The percentage of IL-17 secreting cells in both subsets was affected by the treatment to a lesser extent. Finally, we observed that CD4+ CD25highCD127low regulatory T cells were decreased in MS patients compared to healthy controls and fingolimod significantly increased their frequencies. All together these findings demonstrate that fingolimod functionally modulates the ability of potentially pathogenic effector cells to produce relevant pro-inflammatory cytokines and increases the number of circulating regulatory T cells possibly contributing in restoring a balance between these populations.