High-dimensional analysis of T-cell profiling variations following belimumab treatment in systemic lupus erythematosus.

OBJECTIVE: This study sought to elucidate the molecular impacts of belimumab (BEL) treatment on T-cell immune profiling in SLE. METHODS: We used mass cytometry with 25 marker panels for T-cell immune profiling in peripheral blood T cells (CD3+) from 22 patients with BEL-treated SLE and 20 controls with non-BEL-treated SLE. An unsupervised machine-learning clustering, FlowSOM, was used to identify 39 T-cell clusters (TCLs; TCL01-TCL39). TCLs (% of CD3+) showing significant (p<0.05) associations with BEL treatment (BEL-TCL) were selected by a linear mixed-effects model for comparing groups of time-series data. Furthermore, we analysed the association between BEL treatment and variations in regulatory T-cell (Treg) phenotypes, and the ratio of other T-cell subsets to Treg as secondary analysis. RESULTS: Clinical outcomes: BEL treatment was associated with a decrease in daily prednisolone use (coef=-0.1769, p=0.00074), and an increase in serum CH50 (coef=0.4653, p=0.003), C3 (coef=1.1047, p=0.00001) and C4 (coef=0.2990, p=0.00157) levels. Molecular effects: five distinct BEL-TCLs (TCL 04, 07, 11, 12 and 27) were identified. Among these, BEL-treated patients exhibited increased proportions in the Treg-like cluster TCL11 (coef=0.404, p=0.037) and two naïve TCLs (TCL04 and TCL07). TCL27 showed increased levels (coef=0.222, p=0.037) inversely correlating with baseline C3 levels. Secondary analyses revealed associations between BEL treatment and an increase in Tregs (coef=1.749, p=0.0044), elevated proportions of the fraction of Tregs with inhibitory function (fTregs, coef=0.7294, p=0.0178) and changes in peripheral helper T cells/fTreg (coef=-4.475, p=0.0319) and T helper 17/fTreg ratios (coef=-6.7868, p=0.0327). Additionally, BEL was linked to variations in T-cell immunoglobulin and mucin domain-containing protein-3 expression (coef=0.2422, p=0.039). CONCLUSIONS: The study suggests an association between BEL treatment and variations in T cells, particularly Tregs, in SLE pathologies involving various immune cells.

Abnormally high expression of D1-like dopamine receptors on lupus CD4+ T cells promotes Tfh cell differentiation.

OBJECTIVE: SLE is a chronic autoimmune disease that places a great burden on human society. T follicular helper (Tfh) cells play a critical role in the pathological process of SLE. Therefore, elucidating the mechanism of Tfh cell differentiation will contribute to SLE treatment. Dopamine receptors (DRDs) are members of the family of G protein-coupled receptors and are primarily divided into D1-like and D2-like receptors. Previous studies have found that DRDs can regulate differentiation of immune cells. However, there is currently a lack of research on DRDs and Tfh cells. We here explore the relationship between DRDs and Tfh cells, and analyse the relationship between DRD expression on Tfh cells and the course of SLE. METHODS: We first detected plasma catecholamine concentrations in patients with SLE and healthy controls by mass spectrometry, followed by reverse transcription-quantitative PCR (RT-qPCR) to detect DRD messenger RNA (mRNA) expression in peripheral blood mononuclear cells (PBMCs) and CD4+ T cells, and flow cytometry to detect DRD expression in Tfh cells. Finally, in vitro experiments and RNA sequencing (RNA-seq) were used to explore the possible pathway by which DRDs regulate Tfh cell differentiation. RESULTS: The plasma dopamine concentration in patients with SLE was significantly increased, and abnormal mRNA expression of DRDs was observed in both PBMCs and CD4+ T cells. The results of flow cytometry showed that D1-like receptors were highly expressed in Tfh cells of patients with SLE and associated with disease activity. In vitro induction experiments showed that differentiation of naïve T cells into Tfh cells was accompanied by an increase in D1-like receptor expression. RNA-seq and RT-qPCR results indicate that D1-like receptors might promote Tfh cell differentiation through the Phosphatidylinositol3-kinase (PI3K)/protein kinase B (AKT)/Forkhead box protein O1 (FOXO1)/Kruppel-like factor 2 (Klf2) pathway. CONCLUSION: Tfh cells in patients with SLE highly express D1-like receptors, which correlate with disease activity. D1-like receptors may promote Tfh cell differentiation through the PI3K/AKT/FOXO1/Klf2 pathway.

Lupus pathogenesis and autoimmunity are exacerbated by high fat diet-induced obesity in MRL/lpr mice.

OBJECTIVE: SLE is an autoimmune disease characterised by persistent inflammation and autoantibody production. Genetic predisposition and environmental factors such as a high-fat diet (HFD) may contribute to lupus development. However, the immune cell profile and gender difference in response to HFD in lupus have not been reported. Here we investigated the impact of HFD on lupus pathogenesis and autoimmunity using lupus-prone mice. METHODS: Thirty male and 30 female MRL/lymphoproliferation (lpr) mice were fed with regular diet (RD) or HFD. Body weights were recorded weekly. SLE progression was monitored by skin lesion, urine protein, titres of antidouble-strand DNA (dsDNA) and ANA. At week 14, kidney and skin tissue sections were stained with H&E and periodic acid-Schiff to detect histological kidney index and skin score. Splenocytes were identified by immunofluorescence staining and flow cytometry. RESULTS: HFD significantly increased body weight and lipid levels compared with RD (p<0.01). Skin lesions were observed in 55.6% of the HFD group compared with 11.1% of the RD group, with greater histopathological skin scores in the female HFD group (p<0.01). Although both male and female mice had higher serum IgG in the HFD group than in the RD group, only the male HFD group showed an increased trend in anti-dsDNA Ab and ANA titres. Kidney pathological changes in the HFD group were more severe in male mice than in female mice (p<0.05), detected by proteinuria, kidney index and glomerular cell proliferation. Significant increases of germinal centre B cells and T follicular helper cells were observed in the spleens of HFD mice (p<0.05). CONCLUSION: HFD induced an accelerated and exacerbated lupus development and autoimmunity in MRL/lpr mice. Our results parallel many known clinical lupus phenotypes and sexual dimorphism in which male patients are likelier to have a severe disease (nephritis) than female lupus patients who may have a broader range of lupus symptoms.

Truncated isoforms of GPSM2 containing the GoLoco motif region promote CD4+ T-cell migration in SLE.

OBJECTIVE: SLE is an autoimmune disease with a complex pathogenesis. T-cell infiltration into organs contributes to inflammation and organ damage in SLE. Recently, G-protein signalling modulator 2 (GPSM2) has been shown to be implicated in T-cell migration. METHODS: We analysed the expression levels of GPSM2 and of a truncated isoform of GPSM2 containing the GoLoco motif region in CD4+ T cells from patients with SLE and from healthy individuals by western blot. In a next step, we studied the role of the truncated GPSM2 isoform using a CD4+ T-cell migration assay. RESULTS: Our experiments revealed comparable levels of GPSM2 in CD4+ T cells from patients with SLE and healthy controls. In contrast, the truncated 35 kDa isoform of GPSM2 was significantly more highly expressed in CD4+ T cells from patients with SLE as compared with healthy subjects. Antibody-mediated blockade of the 35 kDa GPSM2 isoform reduced the in vitro capacity of CD4+ T cells to migrate towards the chemokine CCL20. CONCLUSIONS: A truncated GPSM2 isoform containing the GoLoco motif region is upregulated in CD4+ T cells from patients with SLE and promotes CD4+ T-cell migration. Targeting this isoform with specific antibodies might be a promising approach to reduce CD4+ T-cell infiltration into inflamed tissues and to prevent organ damage in SLE.

Peripheral blood T-cell subset and its clinical significance in lupus nephritis patients.

OBJECTIVES: Lupus nephritis (LN) is a common and severe manifestation of SLE. Memory T (TM) cells have been implicated in the pathogenesis of SLE. This study aimed to investigate the clinical significance of T-cell subsets in a cohort of patients with LN. METHOD: The peripheral blood T cells of 24 patients with LN and 13 patients with idiopathic membranous nephropathy (iMN) were analysed by flow cytometry. SLE disease activity was evaluated by SLE Disease Activity Index-2000 (SLEDAI-2K). Patients with LN were followed up for >6 months. RESULTS: Patients with LN presented lower frequency of CD4+ cells and higher percentage of CD8+ cells than patients with iMN, which was primarily due to lower CD4+ cell count. Interestingly, patients with LN under immunosuppressants had lower CD8+CD45RO+ TM frequency (p=0.007), fewer regulatory CD4+ T cells (p=0.04) than those without immunosuppressants. Most CD4+ and CD8+ TM cells in patients with LN showed an effector memory (CD45RO+CCR7+) phenotype. The frequency of CD8+CD45RO+ TM cells among the CD8+ T cells was negatively correlated with white blood cell count, haemoglobin, platelet and serum levels of complements C3 and C4, but was positively correlated with serum IgG, erythrocyte sedimentation rate and SLEDAI score (p<0.05 each). Consistently, the frequency of CD8+CD45RO+ TM cells was higher in patients with LN with positive antidouble-stranded DNA antibody, active renal disease, extrarenal manifestations and with sclerotic glomeruli or moderate-to-severe mesangial hypercellularity in renal pathology (p<0.05). Additionally, CD8+CD45RO+ TM cell frequency was significantly lower in patients with LN with renal complete remission than that in non-remission LN (18.7% vs 31.2%, p<0.05). None of these significant correlations was observed in CD4+ TM cells. CONCLUSION: The frequency of CD8+ TM cells correlates with disease activity and treatment response to immunosuppressant in patients with LN. CD8+ TM monitoring in patients with LN could provide more helpful indices for the monitoring and management of this disease.

Phenotypical changes and clinical significance of CD4+/CD8+ T cells in SLE.

OBJECTIVE: T cells display significant phenotypical changes and play multiple roles in promoting the immune response in SLE. The frequencies of T cell subpopulations in SLE are still not well understood. To better understanding the phenotypic abnormalities of T cells in SLE will help us to clarify disease immunopathology and to find promising biomarkers for disease monitoring and control. METHODS: Peripheral blood CD4+ and CD8+ T cells and their subsets were determined by flow cytometry. Forty-one active SLE patients were selected, including 28 new-onset patients and 13 relapsing patients. One hundred healthy controls (HCs) were enrolled as the control group. The percentages of these cell subsets between patients with SLE and HCs and their relationships with disease activity and autoantibody titers were analysed. Thirteen of 28 new-onset SLE patients were assessed before and after treatment. The changes in the frequencies of these cell subsets and their relationships with renal response were analysed. RESULTS: There was a broad range of anomalies in the proportion of T cell subsets in patients with SLE compared with that of the HCs. Compared with the HCs, a higher frequency of memory T cells and a lower frequency of naïve T cells were noted in patients with SLE. In addition, an imbalance of CD28+ and CD28- cells in CD4+ T cells was observed in patients with SLE. We found that the expanded CD4+CD28- T cells did not decrease after treatment in patients who had impaired renal responses. It was very interesting to exhibit a negative correlation in the frequency between the CD4+CD28- T cells and T regulatory (Treg) cells and a positive correlation between the frequency of CD4+CD28+ T cells and Treg cells in this study. Increased CD8+HLADR+ T cell and CD8+CD38+HLADR+ T cell counts were observed in patients with SLE, suggesting an impaired cytotoxic capacity of CD8+ T cells in SLE. Additionally, we found that CD8+CD38+HLADR+ T cells were closely associated with disease activity, autoantibody titres and renal prognosis. CD4+ CXCR5-PD1+ T cells were expanded in patients with SLE in this study and were associated with disease activity in SLE. Th1 (T helper type 1) cells and Treg cells were decreased, but frequencies of T follicular helper (Tfh) cells, Th2 cells, Th17 cells and Tfh17 cells were increased. A strong correlation between Th17 cells and Tregs with renal involvement was observed in this study. CONCLUSION: The proportions of CD4+CD28- T cells, CD4+CXCR5-PD1+ T cells, CD8+HLADR+ T cells and CD8+CD38+HLADR+ T cells increased in patients with SLE and could be associated with disease activity and renal prognosis.

[How T lymphocytes coordinate rheumatic inflammation]. / Wie T-Lymphozyten rheumatische Entzündung koordinieren.

Helper T (Th) cells play a decisive role in triggering and maintaining chronic rheumatic inflammation. Via secretion of proinflammatory cytokines and expression of costimulatory cell surface molecules, Th lymphocytes coordinate the recruitment and activation of effector cells, which are ultimately responsible for the immunopathology and tissue destruction. However, therapeutic approaches aimed at eliminating Th cells were unsuccessful due to their lack of selectivity. At the German Rheumatism Research Center (Deutsches Rheuma-Forschungszentrum, DRFZ), we are working to improve the understanding of the Th cells involved in chronic inflammatory reactions. Based on this understanding, our aim is to develop novel treatment strategies that selectively target the pathogenic Th lymphocytes causing rheumatic inflammation. The current article summarizes the DRFZ's research activities on this subject.

Analysis of interleukin 1 receptor-associated kinase 1, disease activity and CD4 + T cell subsets in peripheral blood of rheumatoid arthritis patients / 中华风湿病学杂志

Objective:To explore the expression level of interleukin-1 receptor-associated kinase-1 (IRAK1) in the peripheral blood of rheumatoid arthritis (RA) patients and analyze its relevance between disease activity and CD4 + T cell subsets. Methods:① The concentration of IRAK1 in the peripheral blood of 77 RA patients and 24 healthy controls were detected by enzyme linked immunosorbent assay (ELISA). ② The demo-graphic and clinical data of the RA group including disease activity score with 28 joints (DAS28), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), CD4 + T cell subsets in peripheral blood. ③Independent sample t test or Mann-Whitney U test were used to compare the differences between the two groups. Spearman rank correlation test and multiple linear regression were used to analyze the correlation between IRAK1 expression level and clinical data. Results:① The IRAK1 level of the peripheral blood of RA patients was significantly higher than in the normal controls ( P<0.001). ② Compared to normal controls, the peripheral blood of the RA group, the absolute numbers and proportion of regulatory T (Treg) cells were decreased ( P<0.001), the absolute numbers and proportion of helper T (Th) 17 and the ratio of Th17/Treg were increased. Moreover, the ratio of Th17/Treg was also increased. ③ With the increase of disease activity in RA patients, the expression of IRAK1 also increased. The expression of IRAK1 in the peripheral blood of RA group was positively correlated with ESR, number of joints involved and DAS28, and had statistically significant difference between the two groups ( r=0.23, P<0.05; r=0.24, P<0.05; r=0.27, P<0.05). Meanwhile, it was sign-ificantly negatively correlated with the percentage of Treg ( r=-0.27, P<0.05), and was significantly positively correlated with the ratio of Th17/Treg ( r=0.23, P<0.05) . However, there was no significant correlation with the ratio of Th1/Th2( P>0.05). Furthermore, multiple stepwise regression analysis showed that the expression of IRAK1 in the peripheral blood of RA group was positively correlated with ESR and the number of joints involved ( β=0.34, P=0.019; β=0.27, P=0.004), and it was inversely correlated with percentage of Treg ( β=-0.23, P=0.047, R2=0.219). Conclusion:IRAK1 expression in the peripheral blood of RA patients is up-regulated and correlated with disease activity. The decrease of Treg and the imbalance of Th17/Treg caused by high expression of IRAK1 may be one of the main factors for the occurrence and development of RA. Interfering the expression of IRAK1 may be a potential new target for RA treatment.

Changes of humoral and cellular immune indexes in peripheral blood of children with Kawasaki disease at different stages / 中国医师进修杂志

Objective:To investigate the dynamic change characteristics of peripheral blood interferon-γ (INF-γ), interleukin (IL)-4 levels and T helper cell (Th)1/Th2 balance in acute, subacute and restoration stages of children with Kawasaki disease (KD).Methods:Forty-one children with KD received treatment in Women′s and Children′s Hospital Affiliated of School of Medicine University of Electronic Science and Technology of China, Chengdu Women′s and Children′s Central Hospital from May 2017 to January 2020 were enrolled as the observation group, and 41 healthy children examinee from the same period were enrolled as the control group. Children with KD of the observation group were performed with tuberculin pure protein derivative (PPD) test when acute and restoration stage of KD respectively. Peripheral venous blood were drawn from KD children of the observation group in acute, subacute and restoration stage and the control group respectively, serum immune globulin IgG, IgA, IgM and IgE, serum IFN-γ and IL-4 levels were detected by enzyme linked immunosorbent assay (ELISA).Results:Positive rates of PPD test in the restoration stage was higher than that in the acute stage: 65.85%(27/41) vs.17.07%(7/41), there was statistical difference ( χ2 = 20.10, P<0.05). The levels of serum IgG, IgA, IgM and IgE in the acute stage , subacute stage and restoration stagewere gradually decreased ( P<0.05). The levels of serum IgG, IgA, IgM and IgE in the restoration stage and the control group had no significant differences ( P>0.05). The levels of serum IFN-γ and IFN-γ/IL-4 in the acute stage , subacute stage and restoration stage were gradually increased ( P<0.05), the level of IL-4 was gradually decreased ( P<0.05), but the levels of serum IFN-γ, IL-4 and IFN-γ/IL-4 in the restoration stage and the control group had no significant differences ( P>0.05). Conclusions:Among the children with KD in acute stage, serum level of IFN-γ is decreased while serum IL-4 level is increased, and Th1/Th2 balance shifts to Th2. Along with the stabilization of disease, the levels of serum IFN-γ and IL-4 are normalized, and Th1/Th2 balance presents a recovering trend and they almost recover to normal after entering the restoration stage.

Advances on roles of Th22 cells in human common viral infections / 中华临床感染病杂志

Na?ve CD4 + T cells differentiate into a variety of T helper (Th) subsets that secrete various cytokines to exert biological effects. Th22 cells, a novel identified CD4 + T cell subset, are distinct from Th1, Th2 and Th17 cell subsets. Th22 cells express chemokine receptors CCR4, CCR6 and CCR10, and secrete multiple cytokines such as IL-22, IL-13 and TNF-α, but not IL-17, IL-4 IFN-γ; and IL-22 is considered as major effector cytokine of Th22. The understanding on functions and differentiation mechanisms of Th22 cells have been constantly improved, and Th22 cells play important roles in human common viral infections. The article reviews the current advances about the characteristics, function, differentiation of Th22 cells, the roles of Th22 cells and the key molecules in several human common viral infections, which would provide novel immune strategies for the prevention and treatment of human viral infection.

The effect of CD 8+ CD 25+ FoxP3 + regulatory T cell expression levels on balance of helper T cells 1/ helper T cells 2 in peripheral blood of pregnant women with premature rupture of fetal membranes / 中国医师进修杂志

Objective:To investigate the effect of CD 8+ CD 25+ FoxP3 + regulatory T cell (Treg) expression levels in peripheral blood of pregnant women with premature rupture of fetal membranes(PROM) on immune function of helper T cells (Th) 1/Th2. Methods:Thirty cases of pregnant women with PROM (PROM group), 30 cases of normal pregnant women (normal pregnancy group) and 30 cases of normal non-pregnant women (non-pregnancy group) who treated in Binhai County People′s Hospital from September 2019 to May 2020 were collected. Peripheral blood of each group was collected and the proportion of CD 8+ CD 25+ FoxP3 + Treg was determined by flow cytometry. Peripheral blood mononuclear cells (PBMCs) were extracted and FoxP3 mRNA was determined by polymerase chain reaction (PCR). The levels of Th1-related cytokines interferon-γ (IFN-γ), interleukin (IL)-2, and Th2-related cytokines IL-10 and IL-4 were measured by Luminex liquid phase microarray. The effects of CD 8+ CD 25+ FoxP3 + Tregexpression on Th1/Th2 balance were analyzed. Results:The proportion of CD 8+ CD 25+ FoxP3 + Tregand the expression of FoxP3 mRNA in PROM groupand normal pregnancy group were lower than those in non-pregnancy group: (0.15 ± 0.03) %, (0.35 ± 0.09) % vs. (0.47 ± 0.11) %; 0.89 ± 0.11, 3.15 ± 0.67 vs. 3.75 ± 0.23 , the proportion of CD 8+ CD 25+ FoxP3 + Treg and the expression of FoxP3 mRNA in PROM groupwere lower than those in the normal pregnancy group , and the differences were statistically significant ( P<0.05). The levels of Th1-related cytokines IFN-γ and IL-2 in PROM group and normal pregnancy group were higher than those in non-pregnancy group, the level of Th2-related cytokines IL-4 was lower than that in non-pregnancy group , the levels of IFN-γ and IL-2 in PROM group were higher than those in normal pregnancy group, the level of IL-4 was lower than that in normal pregnancy group , and the differences were statistically significant ( P<0.05). In PROM group, the proportion of CD 8+ CD 25+ FoxP3 + Treg and the expression of FoxP3 mRNA in peripheral blood were negatively correlated with Th1-related cytokines IFN-γ ( r = - 0.413, -0.451, P<0.05) and IL-22 ( r = -0.645, -0.535, P<0.05), and were positively correlated with Th2-related cytokines IL-4 ( r = 0.558, 0.469, P<0.05). Conclusions:The proportion of CD 8+ CD 25+ FoxP3 + Treg in peripheral blood of pregnant women with PROM is lower, and the expression level of related FoxP3 mRNA is lower, which all affecte the Th1/Th2 immune balance and cause Th1 immune drift, which may be the related immune mechanism of PROM.

Effectiveness of Xiaoyin Jiedu granules in the treatment of psoriasis vulgaris in patients with blood-heat symptom patterns in terms of Traditional Chinese Medicine.

OBJECTIVE: To evaluate the effectiveness of Xiaoyin Jiedu (XYJD) granules in the treatment of psoriasis vulgaris (PSV) in patients with a blood-heat pattern (BHP) in terms of Traditional Chinese Medicine (TCM). We also aimed to identify the possible underlying immunological mechanism. METHODS: Twenty-five PSV patients with BHP and ten normal controls were enrolled from January 1, 2015 to December 31, 2016. Patients were randomly assigned to either the XYJD group (15 cases) or the placebo group (10 cases), in which patients were treated with XYJD granules or a placebo, respectively. Additionally, albolene was used to relieve skin dryness in these two groups. The psoriasis area and severity indexes, dermatology life quality indexes and itching scores were assessed at the end of the 2nd, 4th and 8th week of treatment. The number of peripheral blood T helper (Th) 9, Th17 and regulatory T cells (Tregs) and the mRNA and protein expression levels of PU.1, RAR-related orphan receptor (ROR)-γt, forkhead box protein 3 (Foxp3), interleukin (IL)-9, IL-17, IL-23 and IL-10 in the control and experimental groups were compared before and after treatment. RESULTS: Psoriasis area and severity indexes and itching scores of patients in the XYJD group were significantly lower than those in the placebo group, whereas dermatology life quality indexes were significantly higher. In comparison with the placebo group, XYJD granules significantly reduced the number of Th17 cells and the mRNA and protein expression levels of Th17-related ROR-γt, IL-17, IL-22 and IL-23 in the peripheral blood and reduced the number of Th9 cells and the mRNA and protein expression levels of Th9-related PU.1 and IL-9. CONCLUSION: XYJD granules were effective against PSV in patients with BHP by reducing the number of Th9 and Th17 cells and the levels of their related cytokines.

Endothelial Cell-Derived Interleukin-18 Released During Ischemia Reperfusion Injury Selectively Expands T Peripheral Helper Cells to Promote Alloantibody Production.

BACKGROUND: Ischemia reperfusion injury (IRI) predisposes to the formation of donor-specific antibodies, a factor contributing to chronic rejection and late allograft loss. METHODS: We describe a mechanism underlying the correlative association between IRI and donor-specific antibodies by using humanized models and patient specimens. RESULTS: IRI induces immunoglobulin M-dependent complement activation on endothelial cells that assembles an NLRP3 (NOD-like receptor pyrin domain-containing protein 3) inflammasome via a Rab5-ZFYVE21-NIK axis and upregulates ICOS-L (inducible costimulator ligand) and PD-L2 (programmed death ligand 2). Endothelial cell-derived interleukin-18 (IL-18) selectively expands a T-cell population (CD4+CD45RO+PD-1hiICOS+CCR2+CXCR5-) displaying features of recently described T peripheral helper cells. This population highly expressed IL-18R1 and promoted donor-specific antibodies in response to IL-18 in vivo. In patients with delayed graft function, a clinical manifestation of IRI, these cells were Ki-67+IL-18R1+ and could be expanded ex vivo in response to IL-18. CONCLUSIONS: IRI promotes elaboration of IL-18 from endothelial cells to selectively expand alloreactive IL-18R1+ T peripheral helper cells in allograft tissues to promote donor-specific antibody formation.

Effect of Tripterygium Glycosides on Differentiation and Balancing of Th17/Treg Cells in Rats With Experimental Colitis / 胃肠病学

Background: Immune factors play an important role in the pathogenesis of inflammatory bowel disease (IBD). Clinical studies have shown that tripterygium glycosides is effective for the treatment of IBD. Aims: To investigate the effect of tripterygium glycosides on differentiation and balancing of Th17/Treg cells in rats with experimental colitis. Methods: Experimental colitis was induced by TNBS-ethanol method in rats to evaluate the therapeutic effect of tripterygium glycosides. After intragastrically administered with normal saline (model group), tripterygium glycosides or mesalazine, respectively once a day for two weeks, the disease activity index (DAI) was assessed, and the colonic mucosal injury was examined macro- and microscopically. Mononuclear cells of mesenteric lymph nodes were extracted, and the levels of Th17/Treg-related cytokines in the supernatant were detected by ELISA method. The expression of proinflammatory cytokines in colon tissues was detected by immunohistochemistry. Results: The symptoms of experimental colitis were more severe in model group. DAI, gross morphological and histopathological score of colonic mucosal injury were significantly higher in model group than in tripterygium glycosides and mesalazine groups (P0.05). Compared with the model group, the levels of IL-23 and TNF-α in the supernatant of mesenteric lymph nodes mononuclear cells in mesalazine group, and the levels of IL-23, TNF-α and IL-6 in tripterygium glycosides group were significantly reduced (P0.05). In rats treated with mesalazine, the expression of IL-6 in colon tissues was down-regulated significantly (P<0.05). Conclusions: Tripterygium glycosides have the potential to inhibit the differentiation of Th17 cells and promote the differentiation of Treg cells in IBD. Regulating the imbalance of Th17/Treg cells might be one of the mechanisms of its therapeutic effect on IBD.

[Effect of cigarette smoke exposure on the expression of CCR7 and levels of Th1/Th2 cytokines in asthmatic rats].

Objective: To investigate the effect of cigarette smoke exposure on the expression of CC Chemokine receptor 7 (CCR7) and levels of Th1/Th2 cytokines in asthmatic rats. Methods: Forty Wistar rats were randomly divided into four groups: control group, asthma group, smoke exposure group, asthma-smoke exposure group. The asthma group were sensitized with ovalbumin (OVA) and Aluminum hydroxide at day 1, 8 and challenged with OVA at day 15 by atomization for 8 weeks.While control group was sensitized and challenged with normal saline instead of OVA.The smoke exposure group was sensitized and challenged with normal saline instead of OVA followed passive smoking for 8 weeks. The asthma-smoke exposure group was challenged with OVA followed passive smoking. The pathological changes of different groups were observed by HE-staining. CCR7 was semiquantitatively analyzed in lungs by immunohistochemistry.The concentration of CC chemokine ligand (CCL)19, CCL21, interferon (IFN)-γ and interleukin (IL)-4 in peripheral blood and CCL19 and CCL21 in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent (ELESA) assay. Results: In asthma group, smoke exposure group and asthma-smoke exposure group, the various degrees of inflammatory reaction appeared in lung tissue and the asthma-smoke exposure group was with the most significant reaction. In the lung tissues of the rats from asthma group, smoke exposure group and asthma-smoke exposure group, the average optical density (AOD) of CCR7 were significantly higher than those in control group (0.350±0.023, 0.252±0.022, 0.400±0.029 vs 0.180±0.020, all P<0.01). The AOD of CCR7 of asthma-smoke exposure group was much higher than both that in asthma group and in smoke exposure group (both P<0.01). In asthma group, smoke exposure group and asthma-smoke exposure group, the concentrations of both CCL19 and CCL21 in peripheral blood and BALF were significantly higher than that in control group (all P<0.01). The concentrations of both CCL19 and CCL21 in peripheral blood and BALF of asthma-smoke exposure group were significantly higher than the results in asthma group and in smoke exposure group (all P<0.01). The concentrations of IFN-γ in peripheral blood of asthma group and asthma-smoke exposure group were lower than those in control group [(33±3), (17±3) vs (70±4) pg/ml], but asthma-smoke exposure group was much lower than the results in asthma group (all P<0.01). The concentration of IFN-γ in peripheral blood of smoke exposure group[(100±5)pg/ml]was higher than that in control group and asthma-smoke exposure group (both P<0.01). In asthma group, smoke exposure group, asthma-smoke exposure group, the concentrations of IL-4 in peripheral blood were significantly higher than those in control group [(54±4), (42±4), (76±4) vs (30±4) pg/ml, all P<0.01]. The concentrations of IL-4 in peripheral blood of asthma-smoke exposure group was significantly higher than those in asthma group and in smoke exposure group (both P<0.01). Conclusion: Cigarette smoke could enhance the expression of CCR7 and its ligand, and it can also result in exacerbations of asthma by reducing the expression level of IFN-γ (the representative of Th1 cytokine) and increasing the expression level of IL-4 (the representative of Th2 cytokine).

[Pathogenesis of atopic dermatitis]. / Pathogenese des atopischen Ekzems.

Atopic dermatitis (AD) is one of the most common chronic inflammatory diseases and is also one of the most frequent reasons to consult a dermatologist. Over the past few years there has been a rapidly growing understanding of the cellular, molecular and immunological relationships as well as genetic variations, which leads to a better comprehension of the disease. Consequently, there are innovative targeted therapies in clinical studies or already approved for therapy. To make reasonable use of the new targeted therapies a good understanding of the pathogenesis is very important. In the future, stratification of patients with AD and the resulting personalized therapies will gain in importance. This review depicts the up to date state of knowledge on the complex pathogenesis of AD.

The expression and significance of Th17 cell/regulatory T cells in peripheral blood of patients with idiopathic inflammatory myopathy / 中华风湿病学杂志

Objective To explore the expression and their significance of peripheral Th17 cells and regulatory T cells (Tregs) in idiopathic inflammatory myopathy,and analyze the relationship between the expression and clinical indicators,imaging and pathological changes.Methods Clinical data,laboratory tests,imaging and pathological changes of IIM cases (n=85) and healthy controls (n=70) were enrolled.Clinical data included the classification,age,gender,course of the disease;laboratory tests including erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),creatine kinase (CK),creatine kinase isoenzyme-MB (CKMB),lactate dehydrogenase (LDH),hydroxybutyrate dehydrogenase (HBDH).The level of peripheral Th17,Treg cells and clinical indicators,laboratory tests,imaging and pathological changes were analyzed retrospectively.Since the data was disregarded from the normal distribution,the median four quantile method was used for statistical description.Two samples were compared with Mann-Whitney U test,and the correlation between variables was Spearman rank correlation analysis.Results ①) The levels of Th17 cells in the case group was not significantly different from that in the control group [6.18(3.42,13.65) cell/μl vs 7.42(5.02,11.13) cell/μl,P＞0.05],the levels of Treg cells in patients was significantly lower than that in the control group [21.25(12.48,35.67) cell/μl vs 36.95(30.37,47.12) cell/μl,P＜0.05],the ratio of Th17/Treg was also significantly higher than that in the control group [0.31(0.21,0.47) vs 0.18(0.14,0.31),P＜0.05].② Peripheral Treg cells levels were not correlated with ESR,CRP,CK-MB,LDH and HBDH (P＞0.05).Peripheral Treg cells levels were negatively correlated with CRP (r=-0.279,P＜0.05),but no correlated with ESR,CK-MB,LDH and HBDH (P＞0.05).③ According to the involvement of important organs,patients were classified into two groups:organ involvement group and non-organ involvement group.The levels of Treg cells in the organ involvement group was fewer than that in non-organ involvement group [16.54(8.84,27.34) cell/ul vs 24.87(14.44,43.37) cell/ul,P＜0.05],and the ratio of Th17/Treg in the organ involvement group was significantly higher than that in non-organ involvement group [0.41(0.29,0.68) vs 0.29(0.19,0.39),P＜0.05].④) Peripheral Th17 cells levels in patients with skeletal muscle inflammatory edema was significantly higher than that of non-inflammatory edema patients [10.70 (4.11,14.51) cell/μl vs 3.10 (1.27,5.15) cell/μl,Z=-2.460,P＜0.05].⑤ The levels of Th17,Treg cells and ratio of Th17/Treg did not correlate with pathological features of inflammatory infiltration (P＞0.05).Conclusion The absolute number of peripheral Treg cells decreases significantly in IIM,and correlates with CRP.Patients with organ involvement have fewer Treg cells,and there is imbalance between Th17 and Treg.When muscle MRI presents with inflammatory edema,patients may have high level of Th17 cells.Our results suggest that Treg cells may play an important role in the pathogenesis of IIM.

Effect of placental protein 14 on the proliferation and differentiation of B cells / 中华风湿病学杂志

Objective To investigate the effect and mechanism of placental protein 14 on the proliferation and differentiation of B cells.Methods The lymphocyte of human peripheral blood was separated by gradient centrifugation.Flow cytometry was used to detect the proportion of CD4+CXCR5+follicular helper T cells (Tfh cells),CD4+CXCRS+Foxp3+ follicular regulatory T cells (Tfr cells),CD3-CD19+B cells and CD3-CD38+ plasma cells.ELISA method was used to detect the concentration of IL-21,IL-10 and TGF-beta in the supernatant,and the co-culture of cells was performed by Transwell chamber;t test was used for comparison between groups.Results The proportion of Tfh cells and Tfr cells in the control group was (2.52±0.16)％ and (1.26±0.24)％,respectively,and that of the placental protein 14 groups were (0.84±0.09)％ and (4.64±0.68)％,respectively.There was a significant difference between the two groups (t=9.150,P=0.000 8 and t=4.669,P=0.009 5).Pplacental protein 14 could further inhibit the secretion of IL-21 (t=5.086,P=0.007 1),and promote the increase of IL-10 and TGF-β concentration (t=3.599,P=0.022 8 and t=6.651,P=0.002 7).The percentage of B cells and plasma cells in the placental protein 14 group were (4.87±0.20)％ and (5.41±0.54)％,which were significantly different from those in the Tfh cell group (t=4.997,P=0.007 5;t=5.110,P=0.006 9).Conclusion Placental protein 14 can inhibit the proliferation of B cells and differentiate into plasma cells by inhibiting the differentiation of Tfh cells and increasing the proportion of Tfr cells.

[Effects of human mesenchymal stem cells on airway inflammation in allergic asthma mice and the underlying mechanism].

Objectives: To investigate the effects of human umbilical cord mesenchymal stem cells (hUC-MSCs) on airway inflammation in an ovalbumin (OVA) induced asthma mouse model and the underlying mechanism. Methods: Twenty-four BALB/c mice were randomly divided into four equal groups: normal control group, OVA-induced asthmatic model group, hUC-MSCs treated group (50 µl of hUC-MSCs was transplanted into the trachea of asthmatic mice ) and hUC-MSCs control group (50 µl of hUC-MSCs was transplanted into the trachea of control mice). Human umbilical cord mesenchymal stem cells from umbilical cord of healthy new born babies were used as the source of hUC-MSCs for this study. The asthmatic conditions of the airways and the lungs were assessed by examining: (1) histopathological changes of the airways and the lungs; (2) expression of cytokines IL-6 and TGF-ß mRNA by real-time PCR; (3) total leukocytes and mast cell count in bronchoalveolar lavage fluid (BALF) and number of IL-17-expressing CD4(+) cells (Th17 cells) in the lung tissue using flow cytometry. Results: Typical histopathological changes of asthma were confirmed in the asthmatic model group. These changes included intensive inflammatory cell infiltration around the airways and patchy airway occlusion by hyperviscous mucus. The number of total leukocytes and mast cells in BALF were significantly increased in the asthmatic mice when compared with the control group (P<0.05). Mice in the asthmatic model group had significantly higher percentage of Th17 cells in lung tissues when compared with the control group (2.90% vs 0.76%, P<0.05). In contrast, in the asthmatic mice treated with hUC-MSCs, the inflammatory cell infiltration was significantly reduced compared with asthmatic mice, as observed by significantly lower leukocytes and mast cells in BALF (P<0.05) and significant reduction in the percentage of Th17 cells in the lung of OVA-challenged mice following hUC-MSCs treatment (percentage of Th17 cells: 0.24% vs 2.90%, P<0.05). The expression of mRNA for IL-6 and TGF-ß was significantly suppressed in the hUC-MSCs treatment group (0.23 vs 2.30 and 0.56 vs 6.60, both P<0.01). No asthmatic pathological changes in both normal and hUC-MSCs control groups were observed. Conclusions: hUC-MSCs significantly inhibit the airway inflammation in OVA-induced asthmatic mice. This inhibition is associated with the suppression of Th17 cells and the down-regulation of inflammatory factors such as IL-6 and TGF-ß in the lungs.

Expression mechanism of helper T cells in children with aplastic anemia pathogenesis / 中国基层医药

Objective To investigate the expression mechanism and significane of helper T cells in children with aplastic anemia(AA).Methods 53 children of AA were divided into heavy AA group(SAA group,21 cases),light AA group(MAA group,i7 cases),AA remission group (CR group,15 cases) by the disease,the other taken the non-aplastic anemia blood platelet disorders caused by reduced or anemia in children (AL group) of 16 cases,20 healthy children were selected as the NC group.The peripheral blood CD4+,CD4+ CD225+,CD4+ CD25+ CD127low,CD4+ IFN-γ(Thl cells),CD4+ IL-4 + (Th2 cells) in each group were detected by flow cytometry.ELISA test was used to detect peripheral blood levels of transforming growth factor-β (TGF-β),interferon-γ (IFN-γ) and interleukin-4 (IL-4).Results The peripheral blood CD4+,CD4+ CD25+,CD4+ CD25+ CD127low,TGF-beta 1 levels in the SAA group were (26.59 ± 4.37) ％,(3.44 ± 0.29) ％,(3.13 ± 1.16) ％,(14.59 ± 3.10) ng/mL,respectively,which in the MAA group were (32.67 ± 3.19) ％,(5.42 ± 0.28) ％,(4.29 ± 1.21) ％,(22.98 ± 3.38) ng/mL,respectively,which in the CR group were (33.13 ±3.24)％,(5.23 ±0.26)％,(4.36 ± 1.33)％,(23.19 ± 3.91) ng/mL,respectively,which in the AL group were (37.98 ± 4.01)％,(6.89 ± 0.28)％,(4.99 ± 1.42)％,(34.46 ± 5.23) ng/mL,respectively,which in the NC group were (38.66 ± 3.41) ％,(7.01 ± 0.38) ％,(5.10 ± 1.52) ％,(35.17 ±5.i4) ng/mL,respectively,the differences among the four groups were statistically significant(F =23.72,25.49,15.24,24.52,all P ＜ 0.05).Peripheral blood Th1,Th2 and Th1/Th2 levels in the SAA group were (13.04 ± 3.01)％,(3.44 ±0.29)％,(1.99 ± 1.17),respectively,which in the MAA group were (11.01 ±2.89)％,(6.28 ±2.99)％,(1.75 ± 0.97),respectively,which in the CR group were (10.38 ± 3.27)％,(6.41 ± 3.18)％,(1.62 ±1.03),respectively,which in the AL group were (8.033 ±.42) ％,(6.35 ± 3.08) ％,(1.26 ± 1.11),respectively,which in the NC group were (8.41 ± 3.84) ％,(6.23 ± 3.44) ％,(1.34 ± 1.12),the differences among the four groups were statistically significant (F =35.92,42.43,22.24,all P ＜ 0.05).Peripheral blood IFN-gamma and IL-4levels in SAA group were (13.04 ± 2.58) pg/mL,(17.22 ± 3.88) pg/mL,respectively,which in MAA group were (10.11 ± 2.22) pg/mL,(17.24 ± 4.21) pg/mL,respectively,which in the CR group were (9.88 ± 2.16) pg/mL,(17.01 ±4.00)pg/mL,respectively,which in the AL group were (8.01 ± 1.68)pg/mL,(16.63 ± 3.58)pg/mL,respectively,which in the NC group were (38.66 ± 3.41) pg/mL,(16.743 ±.81) pg/mL,the differences among the four groups were statistically significant (F =24.17,3.39,all P ＜ 0.05).Conclusion Treg cells decreased,the inhibition extent of TGF-β1 on Th1 cells and IFN-γsecretion decreased,which leading to Th1/Th2 shifted to Th1,leading to hematopoietic marrow failure may be one of the pathogenesis of children sufferred from AA.