Th1 Th2 Research Articles

CD4 T-helper cell cytokine phenotypes and antibody response following tetanus toxoid booster immunization

Routine methods for enumerating antigen-specific T-helper cells may not identify low-frequency phenotypes such as Th2 cells. We compared methods of evaluating such responses to identify tetanus toxoid- (TT) specific Th1, Th2, Th17 and IL10+ cells. Eight healthy subjects were given a TT booster vaccination. Blood was drawn before, 3, 7, 14, and 28days after vaccination and peripheral blood mononuclear cells (PBMC) were cultured for 7days with TT, negative control (diluent), and a positive control (Staphylococcus enterotoxin B [SEB]). Activation markers (CD25 and CD69) were measured after 44h (n=8), cytokines in supernatant after 3 and 7days, and intracellular cytokine staining (ICS) of proliferated cells (identified by dye dilution) after 7days (n=6). Vaccination increased TT-specific expression of CD25 and CD69 on CD3+CD4+ lymphocytes, and TT-specific proliferation at 7, 14 and 28days post vaccination. Vaccination induced TT-specific Th1 (IFN-γ, TNF-α, and IL-2) Th2 (IL-13, IL-5, and IL-4), Th17 (IL-17A) and IL-10+ cells as measured by ICS. TT-specific Th1 cells were the most abundant (12–15% of all TT-specific CD4+ T-cells) while IL10+ (1.8%) Th17 (1.1%) and Th2 cells (0.2–0.6%) were less abundant. TT-specific cytokine concentrations in PBMC supernatants followed the same pattern where a TT-specific IL-9 response was also seen. In conclusion, TT booster vaccination induced a broad T-helper cell response. This method of evaluating cytokine phenotypes may be useful in examining the impact of nutrition and environmental conditions on the plasticity of T-helper cell memory responses.

Regulatory T cells and Th1/Th2 in peripheral blood and their roles in asthmatic children.

To determine the changes in CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg) and the Th1/Th2 ratio in peripheral blood of children with asthma, in order to investigate its association with asthma. A total of 150 children with asthma were allocated into the acute phase group (n=94) and remission phase group (n=56) based on clinical manifestations. The acute phase group was subdivided into the mild group (n=54) and severe group (n=40). Flow cytometry was applied to determine CD4(+)CD25(+)Foxp3(+) Treg, CD4(+)IFN-γ(+) Th1 and CD4(+)IL-4(+) Th2 levels in peripheral blood of different groups, and the results were compared with normal children (control group, n=50). The Treg level was significantly lower in the asthma group than the control group (P<0.01): the Treg level was significantly lower in the acute phase group than the remission phase group and control group (P<0.01) and also significantly lower in the severe group than the mild group (P<0.01). The Th1/Th2 ratio was significantly lower in the asthma group than the control group (P<0.01) and also significantly lower in the acute phase group than the remission phase group and control group (P<0.01). The Treg level in peripheral blood of asthmatic children was negatively correlated with the severity of asthma (r=-0.737, P<0.01) and the Th1/Th2 ratio was also negatively correlated with the severity of asthma (r=-0.615, P<0.01), but the Treg level was positively correlated with the Th1/Th2 ratio (r=0.856, P<0.01). The significantly decreased level of Treg in peripheral blood and Th subset imbalance in asthmatic children suggest the important roles of Treg and Th immunity in pathogenesis of asthma. The Treg level and Th1/Th2 ratio in peripheral blood can be used to evaluate the severity asthma.

Mesenchymal stem cells alleviate TNBS-induced colitis by modulating inflammatory and autoimmune responses

To investigate the potential therapeutic effects of mesenchymal stem cells (MSCs) in inflammatory bowel disease (IBD), we transplanted MSCs into an experimental model of IBD. A rectal enema of trinitrobenzene sulfonic acid (TNBS) (100 mg/kg body weight) was administered to female BALB/c mice. Bone marrow mesenchymal stem cells (BMSCs) were derived from male green fluorescent protein (GFP) transgenic mice and were transplanted intravenously into the experimental animals after disease onset. Clinical activity scores and histological changes were evaluated. GFP and Sex determining region Y gene (SRY) expression were used for cell tracking. Ki67 positive cells and Lgr5-expressing cells were determined to measure proliferative activity. Inflammatory response was determined by measuring the levels of different inflammatory mediators in the colon and serum. The inflammatory cytokines included tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-6, IL-17, IL-4, IL-10, and transforming growth factor (TGF-β). Master regulators of Th1 cells (T-box expressed in T cells, T-bet), Th17 cells (retinoid related orphan receptor gamma(t), RORγt), Th2 cells (GATA family of transcription factors 3, GATA3) and regulatory T cells (forkhead box P3, Foxp3) were also determined. Systemic infusion of GFP-BMSCs ameliorated the clinical and histopathologic severity of colitis, including body weight loss, diarrhea and inflammation, and increased survival (P < 0.05). The cell tracking study showed that MSCs homed to the injured colon. MSCs promoted proliferation of intestinal epithelial cells and differentiation of intestinal stem cells (P < 0.01). This therapeutic effect was mainly mediated by down-regulation of both Th1-Th17-driven autoimmune and inflammatory responses (IL-2, TNF-α, IFN-γ, T-bet; IL-6, IL-17, RORγt), and by up-regulation of Th2 activities (IL-4, IL-10, GATA-3) (P < 0.05). MSCs also induced activated CD4(+)CD25(+)Foxp3(+) regulatory T cells (TGF-β, IL-10, Foxp3) with a suppressive capacity on Th1-Th17 effecter responses and promoted Th2 differentiation in vivo (P < 0.05). MSCs are key regulators of immune and inflammatory responses and may be an attractive candidate for cell-based therapy of IBD.

Benzoxazole Derivative B-98 Ameliorates Dextran Sulfate Sodium-induced Acute Murine Colitis and the Change of T Cell Profiles in Acute Murine Colitis Model

The unique role of enzyme 5-lipoxygenase (5-LO) in the production of leukotrienes makes it a therapeutic target for inflammatory bowel disease (IBD). The aim of this study was to evaluate the effects of B-98, a newly synthesized benzoxazole derivatives and a novel 5-LO inhibitor, in a mouse model of IBD induced by dextran sulfate sodium (DSS). C57BL/6 mice were randomly assigned to four groups: normal control, DSS colitis (DSS+saline), low dose B-98 (DSS＋B-98 20 mg/kg) and high dose B-98 (DSS＋B-98 100 mg/kg). B-98 was administered with 3% DSS intraperitoneally. The severity of the colitis was assessed via the disease activity index (DAI), colon length, and histopathologic grading. The production of inflammatory cytokines interleukin (IL)-6 was determined by RT-PCR. Th cells were examined for the proportion of Th1 cell, Th2 cell, Th9 cell, Th17 cell and Treg cell using intracellular cytometry. The B-98 group showed lower DAI, less shortening of the colon length and lower histopathologic grading compared with the DSS colitis group (p<0.01). The expression of IL-6 in colonic tissue was significantly lower in the B-98 groups than the DSS colitis group (p<0.05). The cellular profiles revealed that the Th1, Th9 and Th17 cells were increased in the DSS colitis group compared to the B-98 group (p<0.05). Our results suggest that acute intestinal inflammation is reduced in the group treated with B-98 by Th1, Th9 and Th17 involved cellular immunity.

Higher serum CCL17 may be a promising predictor of acute exacerbations in chronic hypersensitivity pneumonitis

BackgroundRecent research has suggested that the Th1 and Th2 chemokine/cytokine axis contributes to the development of chronic hypersensitivity pneumonitis (HP). Acute exacerbations (AE) are significant factors in the prognosis of chronic HP. Little is known, however, about these biomarkers in association with AE in chronic HP patients.MethodsFifty-six patients with chronic HP were evaluated, including 14 patients during episodes of AE. Th1 mediators (C-X-C chemokine ligand [CXCL]10 and interferon [IFN]-γ), Th2 mediators (C-C chemokine ligand [CCL]17, interleukin-4, and interleukin-13), and pro-fibrotic mediator (transforming growth factor [TGF]-β) were measured to evaluate the mediators as predictors of AE. C-C chemokine receptor (CCR)4 (receptor for CCL17)-positive lymphocytes were quantified in lung specimens.ResultsSerum CCL17 levels at baseline independently predicted the first episode of AE (HR, 72.0; 95% CI, 5.03-1030.23; p = 0.002). AE was significantly more frequent in the higher-CCL17 group (≥285 pg/ml) than in the lower-CCL17 group (<285 pg/ml) (log-rank test, p = 0.0006; 1-year incidence: higher CCL17 vs. lower CCL17, 14.3% vs. 0.0%). Serum CCL17 levels and CCR4-positive cells during episodes of AE were increased from the baseline (p = 0.01 and 0.031).ConclusionsHigher serum concentrations of CCL17 at baseline may be predictive of AE in patients with chronic HP, and CCL17 may contribute to the pathology of AE by inducing the accumulation of CCR4-positive lymphocytes in the lungs.

Periodontal Disease: Linking the Primary Inflammation to Bone Loss

Periodontal disease (PD), or periodontitis, is defined as a bacterially induced disease of the tooth-supporting (periodontal) tissues. It is characterized by inflammation and bone loss; therefore understanding how they are linked would help to address the most efficacious therapeutic approach. Bacterial infection is the primary etiology but is not sufficient to induce the disease initiation or progression. Indeed, bacteria-derived factors stimulate a local inflammatory reaction and activation of the innate immune system. The innate response involves the recognition of microbial components by host cells, and this event is mediated by toll-like receptors (TLRs) expressed by resident cells and leukocytes. Activation of these cells leads to the release of proinflammatory cytokines and recruitment of phagocytes and lymphocytes. Activation of T and B cells initiates the adaptive immunity with Th1 Th2 Th17 Treg response and antibodies production respectively. In this inflammatory scenario, cytokines involved in bone regulation and maintenance have considerable relevance because tissue destruction is believed to be the consequence of host inflammatory response to the bacterial challenge. In the present review, we summarize host factors including cell populations, cytokines, and mechanisms involved in the destruction of the supporting tissues of the tooth and discuss treatment perspectives based on this knowledge.

Vaccination of Mice with an Antigenic Serine Protease-Like Protein Elicits a Protective Immune Response AgainstTrichinella spiralisInfection

Trichinellosis has major economic impacts on animal husbandry and food safety, and the control and elimination of trichinellosis is a major objective of veterinary medicine. A gene encoding serine protease of Trichinella spiralis (Ts-Adsp) was identified by immunoscreening an adult T. spiralis cDNA library. In this study, the recombinant Ts-Adsp protein (rTs-Adsp) was cloned and expressed in a prokaryotic expression system and purified by Ni-affinity chromatography. To determine whether the purified rTs-Adsp is a potential vaccine candidate for the control of T. spiralis infection, we immunized BALB/c mice with this protein in combination with an alum adjuvant and subsequently challenged with T. spiralis larvae. The results showed that mice vaccinated with rTs-Adsp exhibited an average reduction in the muscle larvae burden of 46.5% relative to the control group. Immunization with the rTs-Adsp antigen induced both humoral and cellular immune responses, which manifested as elevated specific anti-rTs-Adsp IgG and IgE antibodies and a mixed Th1-Th2 response, as determined by Th1 (IFN-γ and IL-2) and Th2 (IL-4, IL-10, and IL-13) cytokine profiling, with the Th2 predominant. Thus, purified rTs-Adsp is able to limit the invasion of T. spiralis , and this protein could be an effective vaccine candidate for trichinellosis.

Immune Derangements in Patients with Myelofibrosis- the Role of Treg, Th17, and sIL2α

Abstract 1763Primary myelofibrosis (PMF), myelofibrosis post essential thrombocytosis (MF-ET), and myelofibrosis post polycythemia vera (MF-PV) have been reported to be associated with autoimmune phenomena, such as Coombs positive anemia, lupus anticoagulant, positive ANA and the presence of circulating immune complex, etc. Regulatory T cells (Treg) and IL-17 producing cells (Th17) have been known to play important roles in modulating immune responses. Hence we studied Treg cells in 38 patients with MF including PMF (25), MF-ET (8), MF-PV (5) and compared them to other MPD patients including ET (7), PV (14), and normal controls (16). Patients on lenalidomide or Pomalidomide were excluded. Blood ( 106 MNCs) were stained for flow cytometry analysis using the Treg Detection Kit (Miltenyi Biotec). The number of Treg cells was calculated as the percentage of positive CD4+ CD25+ FoxP3+ T cells from the number of gated CD4+ cells. Treg function was evaluated by XTT cell proliferation assay (Invitrogen) with ratios of Treg to T-effector cells at 1:1 in the presence of anti-CD3 and CD28 micro-beads (Invitrogen). The results (mean + SE) showed numbers of Treg cells in MF were 0.79 + 0.080, other MPD were 1.27 + 0.20, and normal controls were 1.21 + 0.30. No significant difference was found among the three groups. Treg function was evaluated in MF (18 patients). MPD (21) and controls (16). No significant difference was found among the three groups. Th17 cell assays were performed by culturing Blood CD4+ cells in IMDM medium and stimulated with PMA (25ng/ml), ionomycin (1 ug/ml ) and monensin ( 500ng/ml) for 4h at 37°C with 5% CO2. Then Th17 was stained with Th17 Flow Kit (Biolegend) and analyzed by flow cytometry. The results were expressed as % of isolated CD4+ cells (mean ± SE) as follows: MF (2.31 ± 0.65) (n=15), MPD (1.31 ± 0.32) (n=7), and controls (1.89 ± 0.44) (n=10). No significant difference was found among the three groups. We further studied the soluble interleukin 2 alpha subunit (sIL2α) levels. sIL2 α were measured in plasma by ELISA kits, results were expressed as (mean± SE ) ( pg/ml) as follows: MF ( 3534± 298) (n=18), MPD ( 2303 ± 171) (n=22) and controls ( 1734 ± 115) (n=16), P values were <0.05 in MF vs MPD and MF vs Controls, and P=NS in MPD Vs controls. These results confirm our previous observation (Br J Haematol,1994), that sIL2 α levels were significantly elevated in patients with MF, compared with MPD patients and controls.We further studied the effects of the sIL2α on the immune function in MF patients:A) Effects on the Th1, Th17, and Treg cells. CD4+ cells after isolation were cultured for 7 days in IMDM containing IL-2 (100ug/ml) and anti- CD3CD28 micro-beads, with or without recombinant sIL-2Rα (100ng/ml). The cells then were stimulated with PMA (20ng/ml) ionomycin (1ug/ml) and monensin (1uM) for 4 hours before harvest. Then Th1 Th17 and Treg were numerated by flow cytometry. The results showed no difference in Th1 and Th17 cells in cultures with and without sIL2α, but sIL2α significantly increased the numbers of T reg (1.71 +.28,P=0.02) ( fold increase).B) Effects on the Treg function. Viable CD4+CD25− (106) cells were labeled with CFSE ( Invitrogen), then added unlabeled Tregs at 1:1 (responder: Treg) ratio and stimulated with 50 μl of anti CD3/CD28 micro-beads with and without sIL2α for 4 –7days at 37 °C in a 5% CO2 incubator. Cell proliferation was measured by counting the percentage of CFSEdim cells. The results showed that sIL2α significantly increased the CFSE dim cells. Therefore sIL2α suppressd the T reg cell function and increase the T responsive cell proliferation.C) Identification of cells producing sIL2α. CD4+, CD14+, CD8+, CD20+, and Treg were isolated using isolation kits (Miltenyi ), then cultured either with PHA or anti CD3/CD28 micro-beads. ELISA assay for sIL2α were then performed on the cultured supernatant. The results (mean ± SE) ( pg/ml) showed that CD4+ cells produced 342 ±152.6; CD8 +,71.6 ± 19.8; Treg, 306.9 ± 53.5, while CD14+ and CD20+ cells produced negligible quantity of sIL2α. Thus Treg cells were the cells predominantly producing sIL2α in patients with MF. We conclude that in patients with MF, numbers of Treg and Th17 cells were not different from controls or other MPD patients, but Treg cells produce significantly increased amount of sIL2α which further inhibits Treg function and results in autoimmune phenomenon observed in patients with MF. Disclosures:No relevant conflicts of interest to declare.

Increased Interleukin-17 Producing Effector T Cells During Early Post- Transplant Period

Background: The change of precise immune profile during early post-transplant period has not been fully investigated. In present study, we investigated the response of immune cell subset during early post-transplant period in renal transplant recipients. Methods: Twenty-nine living donor renal transplant recipients were enrolled in this study. We used triple immune suppressant composed of tacrolimus, mycophenolate mofetil and steroid and performed induction therapy by basilliximab in those patients. We prospectively investigated the immune cell profile before and at 1 month after transplantation. We investigated the T cell subset and B cell subset using FACS (fluorescent activated cell sorter) analysis before and after transplantation. Results: Mean age at transplantation was 44.7 ± 11.1 years old and HLA mismatch number was 3.2 ± 2.3. The total lymphocyte counts did not differ significantly before and after transplantation (4.7 x 103 /mL vs. 4.4 x 103/mL, P > 0.05). The percentage of Th1 cell (23.7 ± 8.9 % vs. 18.3 ± 8.8 %, P < 0.05) and Th2 cell (3.7 ± 1.8 % vs. 2.4 ± 1.3 %, P < 0.05) in CD4+ T cell significantly decreased after transplantation compared to before transplantation. In contrast, the percentage of Th17 cell did not change after KT (2.2 ± 1.0 %) compared to before KT (1.9 ± 0.7 %, P > 0.05). CD4+CD25+ T cell out of CD4+ T cell did not change significantly before (2.9 ± 1.5 %) and after KT (2.8 vs. 1.9 %, P > 0.05). The percentage of naïve T cell (TNAIVE) in CD4+ T cell (29.5 ± 10.8 % vs. 30.1 ± 9.9 %, P > 0.05) and central memory T cell (44.8 ± 9.9 % vs. 48.0 ± 13.5 %, P > 0.05) did not alter after KT. In contrast, the percentage of effector memory T cell (TEM) significantly decreased after KT (13.2 ± 4.8 %) compared to before KT (18.4 ± 10.4 %, P < 0.05). However, the proportion of IL-17 producing cell out of TEM was significantly increased after KT (4.5 ± 3.3 % vs. 6.7 ± 4.6 %, P < 0.05). In CD8+ T cell, the percentage of TEM significantly decrease as like in CD4+ T cell (18.6 ± 11.8 % vs. 10.3 vs. 9.0 %, P < 0.05), and the proportion of IL-17 producing T cell in TEM showed increasing pattern, even though it did not reach statistical significance (39.7 ± 33.0 % vs. 41.1 ± 24.9 %, P > 0.05). In B cell subset, all of memory B cell (CD19+CD24+CD38low), mature B cell (CD19+CD24+CD38intermediate, immature B cell (CD19+CD24+CD38high) in CD19+ B cell did not show significant change after transplantation (P > 0.05, respectively). Conclusion: In contrast with another helper T cell subset, Th17 cell response did not decrease and rather IL-17 producing TEM cell increased after transplantation. It suggest that current immune suppressant is not enough to suppress allo-immune responses by Th17 cell during early transplant period.

Evaluation of Traditional Medicines III: The Mechanism of Immune Modulation by &lt;i&gt;PHELA&lt;/i&gt;.

PHELA is a herbal traditional medicine that is under development for use as an immune booster in immune compromised individuals. Therefore, the aim of this study was to determine PHELA's mechanism of action by observing for changes in cytokine profiles. Four groups of Sprague Dawley rats (n = 8) were treated daily and separately with normal-saline, cyclosporine-A, PHELA-only and PHELA+ cyclosporine-A. Thereafter, 4 animals from each group were sacrificed after 7 and 14 days of treatment. Serum Th1 cytokines (IL-2, IFN-γ and TNF-α) and Th2 cytokines (IL-4 and IL-10) were measured by ELISA. The concentrations of Th1 cytokines in the PHELA-only treated group were similar to the control group on days 7 and 14. However, the Th1 cytokines were higher in the PHELA+cyclosporine-A treated group compared to cyclosporine-A group, and cyclosporine-A concentrations were similar in both groups. These results show that PHELA did not stimulate Th1 cytokines of a normal immune system but stimulated them when the immune system was suppressed by cyclosporine-A. In conclusion, PHELA is an immune-stimulant to a compromised immune system.

Crucial Role of Gamma Interferon-Producing CD4+Th1 Cells but Dispensable Function of CD8+T Cell, B Cell, Th2, and Th17 Responses in the Control of Brucella melitensis Infection in Mice

Brucella spp. are facultative intracellular bacterial pathogens responsible for brucellosis, a worldwide zoonosis that causes abortion in domestic animals and chronic febrile disease associated with serious complications in humans. There is currently no approved vaccine against human brucellosis, and antibiotic therapy is long and costly. Development of a safe protective vaccine requires a better understanding of the roles played by components of adaptive immunity in the control of Brucella infection. The importance of lymphocyte subsets in the control of Brucella growth has been investigated separately by various research groups and remains unclear or controversial. Here, we used a large panel of genetically deficient mice to compare the importance of B cells, transporter associated with antigen processing (TAP-1), and major histocompatibility complex class II-dependent pathways of antigen presentation as well as T helper 1 (Th1), Th2, and Th17-mediated responses on the immune control of Brucella melitensis 16 M infection. We clearly confirmed the key function played by gamma interferon (IFN-γ)-producing Th1 CD4(+) T cells in the control of B. melitensis infection, whereas IFN-γ-producing CD8(+) T cells or B cell-mediated humoral immunity plays only a modest role in the clearance of bacteria during primary infection. In the presence of a Th1 response, Th2 or Th17 responses do not really develop or play a positive or negative role during the course of B. melitensis infection. On the whole, these results could improve our ability to develop protective vaccines or therapeutic treatments against brucellosis.

Differential strain‐related tissue immune response to sublethal systemic Aspergillus fumigatus infection in mice

Using a nonlethal systemic Aspergillus fumigatus infection, we have recently shown that similarly efficient elimination of fungus from spleens of prototypic Th1 (C57BL/6) and prototypic Th2 (BALB/c) mice is associated with differential immune responses. In light of these data and given the disseminated character of infection, the aim of the present study is to explore whether there are also strain-dependent differences in antifungal responses in peripheral tissues of infected mice. Although similar efficiency of conidia removal was noted in liver and kidneys of both strains, BALB/c mice seemed more prone to tissue injury. Compared with other nonlymphoid organs, lungs proved immunologically the most responsive in systemic aspergillosis. Lower numbers of neutrophils and macrophages in the lungs of infected BALB/c mice, delayed and lower (compared with C57BL/6 mice) expression of their oxidative activity, along with late IFN-γ and upregulated IL-4 production by lung cells might be responsible for slower elimination of A. fumigatus from the lungs of this mouse strain. The data obtained imply that lungs should be viewed as mandatory organ in evaluation of immune-mediated antifungal potential of drugs in models of systemic/disseminated infection and that strain differences noted in tissue responses should be taken into account in these settings.

Dendritic Cell Sphingosine 1-Phosphate Receptor-3 Regulates Th1–Th2 Polarity in Kidney Ischemia–Reperfusion Injury

Dendritic cells (DCs) are central to innate and adaptive immunity of early kidney ischemia-reperfusion injury (IRI), and strategies to alter DC function may provide new therapeutic opportunities. Sphingosine 1-phosphate (S1P) modulates immunity through binding to its receptors (S1P1-5), and protection from kidney IRI occurs in S1P3-deficient mice. Through a series of experiments we determined that this protective effect was owing in part to differences between S1P3-sufficient and -deficient DCs. Mice lacking S1P3 on bone marrow cells were protected from IRI, and S1P3-deficient DCs displayed an immature phenotype. Wild-type (WT) but not S1P3-deficient DCs injected into mice depleted of DCs prior to kidney IR reconstituted injury. Adoptive transfer (i.e., i.v. injection) of glycolipid (Ag)-loaded WT but not S1P3-deficient DCs into WT mice exacerbated IRI, suggesting that WT but not S1P3-deficient DCs activated NKT cells. Whereas WT DC transfers activated the Th1/IFN-γ pathway, S1P3-deficient DCs activated the Th2/IL-4 pathway, and an IL-4-blocking Ab reversed protection from IRI, supporting the concept that IL-4 mediates the protective effect of S1P3-deficient DCs. Administration of S1P3-deficient DCs 7 d prior to or 3 h after IRI protected mice from IRI and suggests their potential use in cell-based therapy. We conclude that absence of DC S1P3 prevents DC maturation and promotes a Th2/IL-4 response. These findings highlight the importance of DC S1P3 in modulating NKT cell function and IRI and support development of selective S1P3 antagonists for tolerizing DCs for cell-based therapy or for systemic administration for the prevention and treatment of IRI and autoimmune diseases.

Prognostic Role of Serum Cytokines in Patients with Nasopharyngeal Carcinoma

Background: Th1/Th2 cytokine network imbalance plays a major role in cancer development and progression. In this study, we aim to evaluate the relationship between those cytokines and clinical outcome in patients with nasopharyngeal carcinoma (NPC). Patients and Methods: The concentrations of Th1 cytokines (IL-2, TNF-α, IFN-γ) and Th2 cytokines (IL-4, IL-5, IL-10) in the serum were examined by Cytometric Bead Array in a total of 80 nasopharyngeal carcinoma patients pre and post treatment. Associations of those cytokines with clinical pathological factors, treatment response, and overall survival were analyzed. Results: Pretreatment serum levels of IL-2 and TNF-α were closely associated with overall survival. Compared to patients with low IL-2 expression, those with high expression had less risk of death (hazard ratio (HR) = 0.31, 95% confidence interval (CI) 0.13–0.75, p = 0.009). In contrast, TNF-α showed opposite effects on overall survival in patients with NPC. Patients with high TNF-α expression had a more than 2-fold increase in risk of death than those with low TNF-α (HR = 2.66, 95% CI 1.04–6.78, p = 0.041). All HRs were adjusted for age, sex, stage, histology, and treatment. Kaplan-Meier survival analysis showed similar survival differences between the 2 groups. Conclusion: Lower serum IL-2 or elevated serum TNF-α concentrations predict an unfavorable prognosis for patients with NPC.

Th17 cells: critical mediators of host responses to burn injury and sepsis.

Th cells have long been recognized as vital components of the adaptive immune system. Until recently, CD3(+)CD4(+) Th cells were divided into cell-mediated Th1 or humoral Th2 responses. However, the Th1-Th2 hypothesis failed to accommodate the more recently described Th17 cells. Today, the major Th cell subsets include Th1, Th2, Th9, Th17, Th22, and Tregs, each of which produce specific effector cytokines under unique transcriptional regulation. Specifically, Th17 cells produce effector cytokines IL-17, IL-21, and IL-22 under the regulation of ROR-γt. Th17 lymphocytes were first described as orchestrators of neutrophil recruitment and activation and as key players in chronic inflammation and autoimmunity. More recent evidence suggest that Th17 lymphocytes and their effector cytokines play a crucial role in maintaining mucosal immunity and barrier integrity, including the skin, lung, and gut. Burn injury induces global changes to the systemic immune response, including suppressed immune function and increased susceptibility to infection. Moreover, burn trauma is associated with remote organ injury. This relationship between burn and remote organ injury supports the hypothesis that immune suppression may facilitate the development of sepsis, systemic inflammatory response syndrome, and multiple organ dysfunction syndrome in critically ill burn patients. Herein, we discuss this emerging adaptive cell subset in critical care settings, including burn injury and clinical sepsis, and highlight the potential therapeutic role of IL-22.

Lymphocytes in obesity-related adipose tissue inflammation

Inflammation in the white adipose tissue (WAT) is considered a major player in the development of insulin resistance. The role of macrophages accumulating in the WAT during obesity, promoting WAT inflammation and insulin resistance is well established. In contrast, less is known about the role of lymphocytes. Recent studies have implicated different lymphocyte subsets in WAT inflammation. For instance, cytotoxic CD8(+) T cells infiltrating the WAT may contribute to the recruitment, differentiation and activation of macrophages. On the other hand, a differential role for CD4(+) Th1 and CD4(+) Th2 cells has been suggested. Levels of WAT regulatory T cells decrease during the course of obesity and may represent a crucial factor for the maintenance of insulin sensitivity. Moreover, activation of natural killer T cells, an innate-like T cell population, which recognises lipid antigens, promotes insulin resistance and WAT inflammation. Finally, B cells may infiltrate WAT very early in response to high-fat feeding and worsen glucose metabolism through modulation of T cells and the production of pathogenic antibodies. These interesting new findings however bear controversies and introduce novel, yet unanswered, questions. Here, we review and discuss the impact of the different lymphocyte subsets in obesity-related WAT inflammation and attempt to identify the open questions to be answered by future studies.

Distinctive Effects of T Cell Subsets in Neuronal Injury Induced by Cocultured Splenocytes In Vitro and by In Vivo Stroke in Mice

T cells and their subsets modulate ischemic brain injury. We studied the effects of the absence of T cell subsets on brain infarction after in vivo stroke and then used an in vitro coculture system of splenocytes and neurons to further identify the roles of T cell subsets in neuronal death. Stroke was induced by middle cerebral artery suture occlusion in mice and infarct sizes were measured 2 days poststroke. Splenocytes were cocultured with neurons, and neuronal survival was measured 3 days later. A deficiency of both T and B cells (severe combined immunodeficiency) and the paucity of CD4 or CD8 T cells equally resulted in smaller infarct sizes as measured 2 days poststroke. Although a functional deficiency of regulatory T cells had no effect, impaired Th1 immunity reduced infarction and impaired Th2 immunity aggravated brain injury, which may be due to an inhibited and enhanced inflammatory response in mice deficient in Th1 and Th2 immunity, respectively. In the in vitro coculture system, wild-type splenocytes resulted in dose-dependent neuronal death. The neurotoxicity of splenocytes from these immunodeficient mice was consistent with their effects on stroke in vivo, except for the mice with the paucity of CD4 or CD8 T cells, which did not alter the ratio of neuronal death. T cell subsets play critical roles in brain injury induced by stroke. The detrimental versus beneficial effects of Th1 cells and Th2 cells both in vivo and in vitro reveal differential therapeutic target strategies for stroke treatment.

Antiinflammatory effect of androgen receptor activation in human benign prostatic hyperplasia cells

Progression of benign prostatic hyperplasia (BPH) involves chronic inflammation and immune dysregulation. Preclinical studies have demonstrated that prostate inflammation and tissue remodeling are exacerbated by hypogonadism and prevented by testosterone supplementation. We now investigated whether, in humans, hypogonadism was associated with more severe BPH inflammation and the in vitro effect of the selective androgen receptor agonist dihydrotestosterone (DHT) on cultures of stromal cells derived from BPH patients (hBPH). Histological analysis of inflammatory infiltrates in prostatectomy specimens from a cohort of BPH patients and correlation with serum testosterone level was performed. Even after adjusting for confounding factors, hypogonadism was associated with a fivefold increased risk of intraprostatic inflammation, which was also more severe than that observed in eugonadal BPH patients. Triggering hBPH cells by inflammatory stimuli (tumor necrosis factor α, lipopolysaccharide, or CD4(+)T cells) induced abundant secretion of inflammatory/growth factors (interleukin 6 (IL6), IL8, and basic fibroblast growth factor (bFGF)). Co-culture of CD4(+)T cells with hBPH cells induced secretion of Th1 inducer (IL12), Th1-recruiting chemokine (interferon γ inducible protein 10, IP10), and Th2 (IL9)- and Th17 (IL17)-specific cytokines. Pretreatment with DHT inhibited NF-κB activation and suppressed secretion of several inflammatory/growth factors, with the most pronounced effects on IL8, IL6, and bFGF. Reduced inflammatory cytokine production by T-cells, an increase in IL10, and a significant reduction of T cells proliferation suggested that DHT exerted a broad anti inflammatory effect on testosterone cells [corrected]. In conclusion, our data demonstrate that DHT exerts an immune regulatory role on human prostatic stromal cells, inhibiting their potential to actively induce and/or sustain autoimmune and inflammatory responses.

Deficient Regulatory T Cell Activity and Low Frequency of IL-17-Producing T Cells Correlate with the Extent of Cardiomyopathy in Human Chagas' Disease

BackgroundMyocardium damage during Chagas' disease results from the immunological imbalance between pro- and production of anti-inflammatory cytokines and has been explained based on the Th1–Th2 dichotomy and regulatory T cell activity. Recently, we demonstrated that IL-17 produced during experimental T. cruzi infection regulates Th1 cells differentiation and parasite induced myocarditis. Here, we investigated the role of IL-17 and regulatory T cell during human Chagas' disease.Methodology/Principal FindingsFirst, we observed CD4+IL-17+ T cells in culture of peripheral blood mononuclear cells (PBMC) from Chagas' disease patients and we evaluated Th1, Th2, Th17 cytokine profile production in the PBMC cells from Chagas' disease patients (cardiomyopathy-free, and with mild, moderate or severe cardiomyopathy) cultured with T. cruzi antigen. Cultures of PBMC from patients with moderate and severe cardiomyopathy produced high levels of TNF-α, IFN-γ and low levels of IL-10, when compared to mild cardiomyopathy or cardiomyopathy-free patients. Flow cytometry analysis showed higher CD4+IL-17+ cells in PBMC cultured from patients without or with mild cardiomyopathy, in comparison to patients with moderate or severe cardiomyopathy. We then analyzed the presence and function of regulatory T cells in all patients. All groups of Chagas' disease patients presented the same frequency of CD4+CD25+ regulatory T cells. However, CD4+CD25+ T cells from patients with mild cardiomyopathy or cardiomyopathy-free showed higher suppressive activity than those with moderate and severe cardiomyopathy. IFN-γ levels during chronic Chagas' disease are inversely correlated to the LVEF (P = 0.007, r = −0.614), while regulatory T cell activity is directly correlated with LVEF (P = 0.022, r = 0.500).Conclusion/SignificanceThese results indicate that reduced production of the cytokines IL-10 and IL-17 in association with high levels of IFN-γ and TNF-α is correlated with the severity of the Chagas' disease cardiomyopathy, and the immunological imbalance observed may be causally related with deficient suppressor activity of regulatory T cells that controls myocardial inflammation.

Induction of Protective Immunity against Eimeria tenella, Eimeria maxima, and Eimeria acervulina Infections Using Dendritic Cell-Derived Exosomes

This study describes a novel immunization strategy against avian coccidiosis using exosomes derived from Eimeria parasite antigen (Ag)-loaded dendritic cells (DCs). Chicken intestinal DCs were isolated and pulsed in vitro with a mixture of sporozoite-extracted Ags from Eimeria tenella, E. maxima, and E. acervulina, and the cell-derived exosomes were isolated. Chickens were nonimmunized or immunized intramuscularly with exosomes and subsequently noninfected or coinfected with E. tenella, E. maxima, and E. acervulina oocysts. Immune parameters compared among the nonimmunized/noninfected, nonimmunized/infected, and immunized/infected groups were the numbers of cells secreting T(h)1 cytokines, T(h)2 cytokines, interleukin-16 (IL-16), and Ag-reactive antibodies in vitro and in vivo readouts of protective immunity against Eimeria infection. Cecal tonsils, Peyer's patches, and spleens of immunized and infected chickens had increased numbers of cells secreting the IL-16 and the T(h)1 cytokines IL-2 and gamma interferon, greater Ag-stimulated proliferative responses, and higher numbers of Ag-reactive IgG- and IgA-producing cells following in vitro stimulation with the sporozoite Ags compared with the nonimmunized/noninfected and nonimmunized/infected controls. In contrast, the numbers of cells secreting the T(h)2 cytokines IL-4 and IL-10 were diminished in immunized and infected chickens compared with the nonimmunized/noninfected and the nonimmunized/infected controls. Chickens immunized with Ag-loaded exosomes and infected in vivo with Eimeria oocysts had increased body weight gains, reduced feed conversion ratios, diminished fecal oocyst shedding, lessened intestinal lesion scores, and reduced mortality compared with the nonimmunized/infected controls. These results suggest that successful field vaccination against avian coccidiosis using exosomes derived from DCs incubated with Ags isolated from Eimeria species may be possible.