B cells present skewed profile and lose the function of supporting T cell inflammation after Roux-en-Y gastric bypass

Bariatric surgeries, including Roux-en-Y gastric bypass (RYGB) are currently the best treatment for obesity and obesity-related comorbidities, such as type 2 diabetes. However, the underlying mechanism of bariatric surgeries is not entirely understood. Further investigations are needed to improve the success rate and achieve sustained health benefits. Given that B cell dysregulation is a critical component of etiology in inflammatory diseases, whereas obesity and type 2 diabetes represent two major inflammatory disorders, we investigated the effect of RYGB on B cell inflammation. We found that B cells after RYGB presented significantly elevated frequency of interleukin (IL)-10-producing cells and reduced frequency of IL-6-producing cells compared to those before RYGB. When grouping B cell subsets into regulatory (secreting IL-10 and transforming growth factor beta [TGF-β]) and effector (secreting IL-2, IL-4, IL-6, IL-12, interferon gamma [IFN-γ] and tumor necrosis factor alpha [TNF-α]) types, we found that after RYGB, the regulatory to effector B cell ratio was significantly increased. Function analyses showed that B cells before RYGB supported IL-17 secretion from T cells whereas these cells after RYGB lost such capacity. B cells after RYGB also gained the capacity to suppress T cell IFN-γ production through TGF-β-mediated effects, a feature not present in B cells before RYGB. Interestingly, the regulatory to effector B cell ratio was directly associated with the reductions in obesity markers following RYGB, such as BMI and fat mass percentage. Together, these results demonstrated a potential mechanism through which RYGB promoted amelioration of obesity and type 2 diabetes.     

CD19+CD20−CD27hi IL‐s10‐producing B cells are overrepresented in R‐CHOP‐treated DLBCL patients in complete remission

Treatment of diffuse large B cell lymphoma (DLBCL) with rituximab, an anti‐CD20 monoclonal antibody, has resulted in significantly improved patient responses with longer event‐free intervals and higher overall survival rates. However, since rituximab depletes all CD20‐expressing cells, including noncancerous B cells, the effects of rituximab on the normal immunity of DLBCL patients under remission need to be examined. Here, we observed that DLBCL patients under remission contained significantly lower frequencies of total B cells, with a significantly overrepresented interleukin (IL)‐10‐producing B cell (B10) population in the peripheral blood. Further examination confirmed that a large fraction of B10 cells was CD20^?CD27^hi plasmablasts, possibly explaining the persistence of B10 cells after R‐CHOP treatment. We also observed that the percentage of B10 cells in DLBCL patients in remission gradually reduced during the first year of achieving complete remission, primarily due to the replenishment of non‐B10 B cells. Despite this, the percentage of B10 cells in DLBCL patients after 1 year of achieving complete remission was still higher than that in controls. CD4⁺ and CD8⁺ T cells cocultured with B10‐enriched B cells secreted significantly lower levels of proinflammatory cytokines IFN‐g and TNF‐a, compared to those incubated with B10‐depleted B cells. Together, our data observed a long‐lasting overrepresentation of B10 cells in DLBCL patients under remission. Whether this change could impact on the overall anti‐tumor immunity during remission requires further studies.     

Regulatory B cell is critical in bone union process through suppressing proinflammatory cytokines and stimulating Foxp3 in Treg cells

Bone fractures may result in delayed union (DU) or non‐union (NU) in some patients. Evidence suggests that the skewing of the immune system toward the proinflammatory type is a contributing factor. Because B cells were previously found to infiltrate the fracture healing site at abundant levels, we examined the regulatory B cells (Bregs) in DU/NU patients. In bone fracture patients with normal healing, the frequency of interleukin (IL)‐10‐expressing B cells was significantly upregulated in the early healing process (6 weeks post‐surgery) and was downregulated later on (18 weeks post‐surgery), whereas in DU/NU patients, the early upregulation of IL‐10‐expressing B cells was missing. The majority of IL‐10‐expressing B cells were concentrated in the IgM⁺CD27⁺ fraction in both controls and patients. IgM⁺CD27⁺ B cells effectively suppressed interferon gamma (IFN‐γ), tumor necrosis factor alpha (TNF‐α), and IL‐2 expression from CD4⁺ T cells, as well as IFN‐γ and TNF‐α expression from CD8⁺ T cells. The IgM⁺CD27⁺ B cell‐mediated suppression was restricted to the sample from the early healing time point in controls, as the IgM⁺CD27⁺ B cells from normal healing patients later on or from DU/NU patients did not present significant regulatory function. In addition, culturing of CD4⁺CD25⁺ Tregs with IgM⁺CD27⁺ B cells from controls at early healing time point resulted in higher Foxp3 expression, a function absent in controls at later time point, or in DU/NU patients. In conclusion, our results support a role of B cell‐mediated regulation early during the bone healing process.     

Function of follicular helper T cell is impaired and correlates with survival time in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) represents one of the most common and aggressive cancers worldwide. The PD-1/PD-L1 interaction plays important roles in cancer immunology, and expression of PD-L1 has been discovered in NSCLC tumor cells. Since follicular helper T (Tfh) cells have characteristic high PD-1 expression, we therefore investigated the inflammatory status of Tfh in NSCLC. CD4⁺ CXCR5⁺ T cell population was examined to define Tfh cells. Data showed that frequency of Tfh cells in peripheral blood was significantly lower in NSCLC patients than in healthy controls. In both primary and metastatic tumors, infiltration of Tfh cells was observed, suggesting that they participated in the antitumor immunity of NSCLC patients. Compared to other T cell subsets, the Tfh cells from the peripheral blood and the resected tumors of NSCLC patients presented elevated apoptosis and reduced proliferation capacity. The Tfh cells from NSCLC patients were also less effective at downregulating IgD and upregulating CD27 expression in naive B cells, and induced less IgM, IgG and IgA secretion, than those from healthy controls. We then found that the survival time from the date of surgery was positively correlated with the frequency of tumor-infiltrating Tfh cells in NSCLC subjects. Overall, the results from this study demonstrated that the Tfh cells were likely involved in the antitumor immunity and were associated with better clinical outcomes, but suffered strong immunosuppression in NSCLC. Enhancing the Tfh cell activity therefore represents a potential therapeutic strategy in NSCLC.     

Analyzing the percentage of different PD‐1+ T cell subsets in peripheral blood and bronchoalveolar lavage fluid of small cell lung cancer patients: A prospective study

The present study was designed to evaluate the percentage of different programmed cell death‐1 (PD‐1)⁺ T cell subsets in peripheral blood and bronchoalveolar lavage fluid (BALF) of small cell lung cancer (SCLC) patients. The percentages of PD‐1⁺ T cell subsets in peripheral blood and BALF samples obtained from 52 lung cancer and 20 pneumonia patients, and 20 healthy controls were examined by flow cytometry. In addition, clinical parameters, such as erythrocyte sedimentation rate (ESR) and C‐reactive protein (CRP) levels, were also determined using Spearman's correlation test to assess their association with PD‐1⁺ T cell subsets. These present results revealed that the percentage of circulating PD‐1⁺ Tfh and peripheral helper T cells (Tph) cells significantly increased in peripheral blood of SCLC patients, when compared to non‐small cell lung cancer (NSCLC) pneumonia patients and healthy controls. In addition, PD‐1⁺ Tfh cells were also significantly enhanced in patients in the extensive‐stage group. In contrast, the BALF samples of SCLC patients exhibited a significant decrease in percentage of Tph cells. An overall imbalance was observed between PD‐1⁺Tfh and Tph cells in both compartments. Furthermore, SCLC patients exhibited a significant decrease in the percentage of circulating PD‐1⁺ Tfh and Tph cells following chemotherapy, and the in vitro analysis revealed that the concentration of IL‐2 and IFN‐γ derived from PD‐1 + Tfh cells in SCLC were significantly lower than that from NSCLC. However, this had no significant correlation with disease severity. The present study indicated that elevated circulating PD‐1⁺ T cells can primarily be used as a biomarker for disease diagnosis and a potential therapeutic target.     

Inhibition of CD8+ T cells and elimination of myeloid cells by CD4+ Foxp3− T regulatory type 1 cells in acute respiratory distress syndrome

Acute lung injury and acute respiratory distress syndrome (ARDS) are caused by rapid‐onset bilateral pulmonary inflammation. We therefore investigated the potential role of interleukin (IL)‐10⁺CD4⁺ Tr1 cells, a regulatory T cell subset with previously identified immunosuppressive functions, in ARDS patients. We first showed that circulating Tr1 cells were upregulated in active and resolved ARDS patients compared to healthy controls and pneumonia patient controls. A significant fraction of these Tr1 cells expressed granzyme B and perforin, while most Tr1 cells did not express interferon gamma (IFN‐γ), IL‐4, IL‐17 or FOXP3, suggesting that the effector functions of these Tr1 cells were primarily mediated by IL‐10, granzyme B, and perforin. Indeed, Tr1 cells effectively suppressed CD8⁺ T cell IFN‐γ production and induced lysis of monocytes and dendritic cells in vitro. The elimination of myeloid antigen‐presenting cells depended on granzyme B production. We also discovered that Tr1 cells could be identified in the bronchoalveolar lavage fluid collected from ARDS patients. All these results suggested that Tr1 cells possessed the capacity to downregulate inflammation in ARDS. In support of this, we found that ARDS patients who resolved the inflammation and survived the syndrome contained significantly higher levels of Tr1 cells than ARDS patients who succumbed to the syndrome. Overall, this report added a novel piece of evidence that ARDS could be intervened by regulatory T cell‐mediated suppressive mechanisms.     

A higher frequency of IL‐10‐producing B cells in Hepatitis B virus‐associated membranous nephropathy

The purpose of this study is to elucidate the potential role of interleukin (IL)‐10⁺ regulatory B cells and other B cell subsets in the development of hepatitis B virus‐associated membranous nephropathy (HBV‐MN). A total of 14 patients with new onset HBV‐MN, 12 individuals with immune‐tolerant HBV infection (HBV‐IT), and 12 healthy controls (HC) were examined for the percentages of CD38⁺, CD86⁺, CD27⁺, CD95⁺ and IL‐10⁺ B cells by flow cytometry. Serum IL‐10 concentration was examined by enzyme‐linked immunosorbent assay (ELISA). The percentages of CD38⁺CD19⁺, CD86⁺CD19⁺, CD38⁺CD86⁺CD19⁺, and CD95⁺CD19⁺ B cells were significantly higher in HBV‐MN patients than the HBV‐IT and HC. The percentages of CD5⁺CD19⁺, IL‐10⁺CD19⁺ B cells and serum IL‐10 level in HBV‐MN patients were significantly higher than the HC, and lower than the HBV‐IT. Percentages of CD38⁺CD19⁺, and CD86⁺CD19⁺ B cells were reduced after treatment, while the percentages of CD5⁺CD1d⁺CD19⁺, CD5⁺CD1d⁺IL‐10⁺CD19⁺, and IL‐10⁺CD19⁺ B cells were increased. The 24 h urinary protein concentration was positively correlated with the percentage of CD38⁺CD19⁺, and negatively correlated with the percentage of IL‐10⁺CD19⁺ B cells and serum IL‐10 level. Similarly, the value of eGFR was negatively correlated with the percentage of CD38⁺CD19⁺, and positively correlated with the percentage of IL‐10⁺CD19⁺ B cells and serum IL‐10 level. Serum IL‐10 level and the percentage of IL‐10⁺CD19⁺ were negatively correlated with the percentages of CD38⁺CD19⁺, and CD86⁺CD19⁺ B cells. These results suggest that CD86⁺CD19⁺, CD38⁺CD86⁺CD19⁺, CD95⁺CD19⁺, and especially CD38⁺CD19⁺ and IL‐10⁺CD19⁺ cells may participate in the pathogenesis of HBV‐MN.     

Increased ratio of ICOS+/PD‐1+ follicular helper T cells positively correlates with the development of human idiopathic membranous nephropathy

To identify the frequencies of different subsets of peripheral blood follicular helper T (Tfh) cells in human idiopathic membranous nephropathy (IMN), 39 patients with new onset IMN and 18 age‐ and gender‐matched healthy controls (HC) were enrolled for this study. The frequency of Tfh cells in venous blood were measured by flow cytometry, while concentration of serum IL‐21 was detected by enzyme‐linked immunosorbent assay. Correlation between the clinical features of IMN and Tfh cells was assessed by Spearman's rank correlation test. Overall, the frequencies of total, ICOS⁺, and PD‐1⁺ Tfh cells were increased in IMN patients, while the ratio of ICOS⁺/PD‐1⁺ Tfh cells positively correlated with IMN progression. However, the elevated serum IL‐21 level in three subgroups of IMN patients, stratified based on 24‐h urine protein levels, was not statistically significant compared to HC. Nonetheless, intracellular IL‐21 in Tfh cells was generally increased in all IMN patients, and closely correlated with IMN development. Finally, the frequency of IL‐21⁺ Tfh cells and the ratio of ICOS⁺/PD‐1⁺ Tfh cells were positively correlated with the estimated 24‐h urine protein of IMN patients. The data indicated that Tfh cells contribute to the pathogenicity of IMN. The ratio of ICOS⁺/PD‐1⁺ Tfh cells and the frequency of IL‐21⁺ Tfh cells may be indicators for evaluating the IMN development.     

Follicular regulatory T cells infiltrated the ovarian carcinoma and resulted in CD8 T cell dysfunction dependent on IL-10 pathway

A high Treg/CD8 T cell ratio in ovarian carcinoma was negatively associated with the prognosis of the patients. The human follicular regulatory T (Tfr) cells are a newly characterized subset of Treg cells with features of both follicular helper T (Tfh) cells (CXCR5⁺) and canonical Treg cells (CD25⁺Foxp3⁺). The role of Tfr cells in ovarian cancer is yet unclear. We found that in peripheral blood, the ovarian cancer patients presented significantly higher levels of both CD4⁺CD25⁺CD127⁻CXCR5⁺ T cells and CD4⁺CD25⁺CD127⁻CXCR5⁺Foxp3⁺ T cells than the healthy controls. In resected tumor samples, Tfr cells represented a much greater percentage of lymphocytes than in peripheral blood. Interestingly, the circulating Tfr cells from ovarian cancer patients presented significantly higher TGFB1 and IL10 expression than their counterparts in healthy controls directly ex vivo, and significantly higher IL10 after stimulation. The tumor-infiltrating Tfr cells presented further upregulated expression of TGFB1 and IL10. In addition, the levels of TGFB1 and IL10 expression by Tfr cells negatively associated with the expression of IFNG in tumor-infiltrating CD8 T cells. In an in vitro CD8 T cell/Tfr cell coculture system, we found that Tfr cells could significantly suppress the activation of CD8 T cells, in a manner that was dependent on IL-10 and probably on TGF-β. Overall, our study found that Tfr cells could suppress CD8 T cells, and in ovarian cancer patients, the Tfr cells were increased in both frequency and function.     

Anti‐inflammatory effects of Artemisia princeps in antigen‐stimulated T cells and regulatory T cells

Objectives The aim was to investigate the anti‐inflammatory effects of Artemisia princeps extract on the activity of anti‐CD3/CD28‐stimulated CD4⁺CD25^‐ T cells and antigen‐expanded regulatory T cells. Methods CD4⁺CD25^‐ T cells were activated with coated anti‐CD3 and anti‐CD28 and cultured in the presence or absence of various concentrations of A. princeps extract. The cultures were pulsed on Day 6 with [³H]thymidine and, after harvesting the cells, [³H] thymidine incorporation was measured. For analysis of interleukin‐2 and interferon‐γ secreted from CD4⁺CD25^‐ T cells, culture supernatants were collected on Days 2 and 6. For the analysis of interleukin‐10 secreted from the CD4⁺CD25^‐ T cells and expanded regulatory T cells, supernatants were collected after 2 and 7 days, respectively. Cytokine levels were determined using an enzyme‐linked immunosorbent assay. Potential medicinal components of the A. princeps extract were determined using gas chromatography–mass spectrometry. Key findings A. princeps (30 μg/ml) effectively suppressed proliferation of CD4⁺CD25^‐ T cells that were stimulated with anti‐CD3/CD28 without causing cytotoxicity in spleen cells incubated under conditions lacking antigen stimulation. A. princeps inhibited production of the pro‐inflammatory cytokines interleukin‐2 and interferon‐γ in anti‐CD3/CD28‐stimulated CD4⁺CD25^‐ T cells. Also, the extract slightly increased production of the anti‐inflammatory cytokine interleukin‐10 in these cells. In regulatory T cells expanded by anti‐CD3/CD28, A. princeps increased production of interleukin‐10 and Foxp3. Conclusions The results suggest that A. princeps may be useful in the treatment of autoimmune diseases and organ transplantation rejection by inhibiting proliferation of inflammatory T cells, suppressing inflammatory processes in antigen‐stimulated CD4⁺CD25^‐ T cells and increasing activity of expanded regulatory T cells.     

The role of interleukin‐4 in rheumatic diseases

Rheumatism is a group of diseases, most of which are autoimmune diseases, that violate joints, bones, muscles, blood vessels and related soft tissue. As is well known, cytokines play a role in the pathogenesis of several rheumatic diseases, such as rheumatoid arthritis, spondyloarthritides, and systemic lupus erythematosus. Recently, the role of interleukin‐4 (IL‐4), which may participate in the mechanism of rheumatism, have been discovered. It is reported that IL‐4 takes part in the regulation of T cell activation, differentiation, proliferation, and survival of different T cell types. IL‐4 also has an immunomodulatory effect on B cells, mast cells, macrophages, and many cell types. A review of the literature on functions of IL‐4 in rheumatic diseases is presented.     

CCR5 plays an important role in resolving an inflammatory response to single‐walled carbon nanotubes

Owing to the development of new materials and technology, the pollutants in the environment are becoming more varied and complex over time. In our previous study using ICR mice, we suggested that a single intratracheal instillation of single‐walled carbon nanotubes (SWCNTs) induced early lung fibrosis and subchronic tissue damage. In the present study, to investigate the role of CCR5 in inflammatory responses to the uptake of SWCNTs, we compared BAL (Bronchoalveolar lavage) cell composition, cell cycles, cytokines, cell phenotypes, inflammatory response‐related proteins, cell surface receptors and histopathology using CCR5 knockout (KO) and wild‐type mice. Results showed that the distribution of neutrophils in BAL fluid significantly decreased in KO mice. The expression of apoptosis‐related proteins including caspase‐3, p53, phospho‐p53, p21 and cleaved PARP, TGF βl and mesothelin markedly increased in KO mice compared with wild‐type mice. Histopathological lesions were also more frequently noted in KO mice. Moreover, the secretion of IL‐13 and IL‐17 with IL‐6 significantly increased in KO mice compared with wild‐type mice, whereas that of IL‐12 significantly decreased in comparison to wild‐type mice. The distribution of B cells and CD8+ T cells was predominant in the inflammatory responses in KO mice, whereas that of T cells and CD4+ T cells was predominant in the inflammatory responses in wild‐type mice. Furthermore, the expression of CCR4 and CCR7 significantly increased in KO mice. Based on these results, we suggest that the absence of CCR5 delays the resolution of inflammatory responses triggered by SWCNTs inflowing into the lungs and shifts inflammatory response for SWCNTs clearance from Th1‐type to Th2‐type. Copyright © 2012 John Wiley & Sons, Ltd.     

PM10‐biogenic fraction drives the seasonal variation of proinflammatory response in A549 cells

PM10 was collected in a Milan urban site, representative of the city air quality, during winter and summer 2006. Mean daily PM10 concentration was 48 μg m^?3 during summer and 148 μg m^?3 during winter. Particles collected on Teflon filters were chemically characterized and the endotoxin content determined by the LAL test. PM10‐induced cell toxicity, assessed with MTT and LDH methods, and proinflammatory potential, monitored by IL‐6 and IL‐8 cytokines release, were investigated on the human alveolar epithelial cell line A549 exposed to increasing doses of PM. Besides untreated cells, exposure to inert carbon particles (2–12 μm) was also used as additional control. Both cell toxicity and proinflammatory potency resulted to be higher for summer PM10 with respect of winter PM10, with IL‐6 showing the highest dose‐dependent release. The relevance of biogenic components adsorbed onto PM10 in eliciting the proinflammatory mediators release was investigated by inhibition experiments. Polymixin B (Poly) was used to inhibit particle‐bind LPS while Toll‐like receptor‐2 antibody (a‐TLR2) to specifically block the activation of this receptor. While cell viability was not modulated in cells coexposed to PM10 and Poly or a‐TLR2 or both, inflammatory response did it, with IL‐6 release being the most inhibited. In conclusion, Milan PM10‐induced seasonal‐dependent biological effects, with summer particles showing higher cytotoxic and proinflammatory potential. Cytotoxicity seemed to be unaffected by the PM biogenic components, while inflammation was significantly reduced after the inhibition of some biogenic activated pathways. Besides, the PM‐associated biogenic activity does not entirely justify the PM‐induced inflammatory effects. © 2010 Wiley Periodicals, Inc. Environ Toxicol 2012.     

Systemic inflammatory response syndrome following burns is mediated by brain natriuretic peptide/natriuretic peptide A receptor‐induced shock factor 1 signaling pathway

The aim of this study was to determine whether systemic inflammatory response syndrome (SIRS) in burn patients is mediated by the brain natriuretic peptide (BNP)/natriuretic peptide A receptor (NPRA)‐induced heat shock factor 1 (HSF‐1) signalling pathway. Mononuclear cells (MNCs) that were isolated from patients with burn injuries and SIRS mouse models and a RAW264.7 cell line were treated with normal serum or serum obtained from animals with burn injuries. In parallel, small hairpin RNAs (shRNAs) against BNP or NPRA were transfected in both cell types. Western blotting (WB) and enzyme‐linked immunosorbent assay (ELISA) were used to detect protein expression and inflammatory factor levels, respectively. We found that interleukin (IL)‐12, tumour necrosis factor (TNF)‐α, C‐reactive protein (CRP), and BNP levels were increased and IL‐10 levels were decreased in the plasma and MNCs in vivo in the animal model of SIRS. Additionally, NPRA was upregulated, whereas HSF‐1 was downregulated in monocytes in vivo. Treatment of RAW264.7 cells with burn serum or BNP induced IL‐12, TNF‐α, and CRP secretion as well as HSF‐1 expression. Finally, silencing BNP with shRNA interrupted the effect of burn serum on RAW264.7 cells, and silencing NPRA blocked burn serum‐ and BNP‐mediated changes in RAW264.7 cells. These results suggest that the interaction of NPRA with BNP secreted from circulatory MNCs as well as mononuclear macrophages leads to inflammation via HSF‐1 during SIRS development following serious burn injury.     

CD4+ T cell imbalance is associated with recurrent endometrial polyps

Endometrial polyps (EPs) are localized benign overgrowths at the endometrium, with currently unknown aetiology and pathogenesis. Although symptoms of EP can be alleviated or resolved by hysteroscopic polypectomy, a significant fraction of individuals develop recurrent EPs after initial EP removal. In rare cases, EPs may also undergo malignant transformation. In‐depth understanding of the mechanisms that are involved in EP development is urgently needed. Recent works indicate that dysregulations in the immune system participate in the development of a variety of symptoms, such as aging, obesity and hypertension, many of which are EP risk factors. Based on these discoveries, we investigated the cellular immune system in premenopausal women with and without EP. Compared to EP‐free controls, the women with EP presented significantly higher RORC expression but unchanged TBX21 and FOXP3 expression in the circulating CD4⁺ T cells. When stimulated with PMA/ionomycin, CD4⁺ T cells from women with EP presented significantly higher interferon (IFN)‐γ and interleukin (IL)‐17 secretion, and lower transforming growth factor (TGF)‐β secretion. Hysteroscopic polypectomy did not significantly alter the composition of CD4⁺ T cells, as the women with EP presented a similar upregulation of Th17 inflammation and a downregulation of regulatory T cell (Treg) response postoperatively. Notably, in women that developed recurrent EP, the CD4⁺ T cells presented higher preoperative and postoperative RORC, IFN‐γ, and IL‐17 expression, as well as lower postoperative FOXP3 and TGF‐β expression, than hysteroscopic polypectomy‐treated women without EP recurrence. These data demonstrated an association between CD4⁺ T cell imbalance and recurrent EP development.     

CP‐25 Alleviates Experimental Sjögren's Syndrome Features in NOD/Ltj Mice and Modulates T Lymphocyte Subsets

Primary Sjögren's syndrome (pSS) is a chronic inflammatory autoimmune illness of the moisture‐producing glands such as salivary glands that is characterized by various immune abnormalities. The aetiology of pSS remains unclear and there is no curative agent. In this study, we investigated the putative therapeutic effects on a NOD/Ltj mouse model of Sjögren's syndrome‐like disorders of an ester derivative of paeoniflorin, paeoniflorin‐6′O‐benzene (termed CP‐25). Our study showed that CP‐25 alleviated effectively clinical manifestations in NOD/Ltj mice resulting, for example, in increased salivary flow and reduced histopathological scores. Furthermore, CP‐25 decreased lymphocyte viability in NOD/Ltj mice and attenuated the infiltration of Th1 cells and Th2 cells into the salivary glands of NOD/Ltj mice. In the spleen on NOD/Ltj mice, CP‐25 skewed the ratio of Th17 and regulatory T cells towards regulatory T cells. After treatment, concentrations of anti‐La/SSB and IgG antibodies were reduced and the titre of the inflammatory cytokines IFN‐γ, IL‐4, IL‐6 and IL‐17A in the serum on NOD/Ltj mice was alleviated. Thus, we define CP‐25 as a novel compound that is a potent therapeutic agent for pSS by modulating T lymphocyte subsets. Future studies will validate the use of CP‐25 as a therapeutic strategy for the treatment of pSS.     

Anti‐inflammatory effects of synthetic compound KT‐14480 in lipopolysaccharide‐stimulated microglia cells

Objectives Neurodegenerative diseases have a prominent inflammatory component. Several synthetic fluorovinyloxyacetamide derivatives were screened by microglia cell‐based assay in order to identify novel compounds that inhibit the inflammatory activation of microglia. Methods Microglia cell‐based nitric oxide assay was employed to screen the compounds. RT‐PCR and ELISA were conducted to evaluate the expression of inflammatory gene expression. Molecular mechanisms were determined by western blot analysis, immunocytochemistry, EMSA, and microglia/neuroblastoma cocultures. Key findings A fluorovinyloxyacetamide compound KT‐14480 significantly suppressed nitric oxide production in lipopolysaccharide‐stimulated microglia cells. KT‐14480 also suppressed the secretion and expression of several inflammatory mediators such as tumour necrosis factor‐α, interleukin‐1β and inducible nitric oxide synthase. Additional studies showed that these inhibitory effects were accompanied by the suppression of nuclear factor‐κB and neuroprotection in the microglia/neuroblastoma coculture. Conclusions Our results indicate that the anti‐inflammatory compound KT‐14480 may be a novel therapeutic drug candidate against neuroinflammatory diseases.     

In vitro stimulatory effect of N‐acetyl tryptophan‐glucopyranoside against gamma radiation induced immunosuppression

Radiation‐induced manifestations like free radical burst, oxidative damage and apoptosis leading to cell death. In present study, N‐acetyl tryptophan glucopyranoside (NATG) was assessed for its immune‐radioprotective activities using J774A.1 cells. Clonogenic cell survival, cell cycle progression and cytokines i.e. IFN‐γ, TNF‐α, IL‐2, IL‐10, IL‐12, IL‐13 and IL‐17A expression were evaluated in irradiated and NATG pretreated cells using clonogenic formation ability, flow cytometry and ELISA assay. Results indicated that 0.25μg/ml NATG exhibited maximum radioprotection against gamma‐radiation (2Gy) without intervening in cell cycle progression. NATG pretreated (−2 h) plus irradiated cells showed significant elevation in IFN‐γ (∼38.2%), IL‐17A (∼53.7%) and IL‐12 (∼58.8%) expression as compared to only irradiated cells. Conversely, significant decrease in TNF‐α (∼21.6%), IL‐10 (∼31.2%), IL‐2 (∼23.7%) and IL‐13 expression (∼17.8%) were observed in NATG pretreated plus irradiated cells as compared to irradiated cells. Conclusively, NATG pretreatment to irradiated J774A.1 cells, stimulate Th₁ while diminish Th₂ cytokines that contributes to radioprotection.