Effect of Sishen Pills and its split prescriptions on Tfr/Tfh9/Tfh17 cells in colitis mice / 中国中药杂志

This study aims to investigate the regulatory effect of Sishen Pills(SSP) and its split prescriptions Ershen Pills(EP) and Wuweizi Powder(WP) on T follicular helper(Tfh) cell subset in the dextran sodium sulfate(DSS)-induced colitis mice and the mechanism. A total of 60 male SPF BALB/c mice were used, 10 of which were randomly selected as the normal group. The rest 50 were induced with 3% DSS solution for colitis modeling. After modeling, they were randomized into 5 groups: model group, SSP group, EP group, WP group, and mesalazine group. Body mass, colon mass, colon mass index, colon length, and unit colon mass index in each group were observed. After hematoxylin-eosin(HE) staining, the pathological injury of colon tissue was scored. The expression levels of molecules related to the STAT/SOCS signaling pathway in colon tissues were analyzed by Western blot. Differentiation levels of Tfh cells such as CD4~+CXCR5~+IL-9~+(Tfh9), CD4~+CXCR5~+IL-17~+(Tfh17), and CD4~+CXCR5~+Foxp3~+(Tfr) in peripheral blood of mice were detected by flow cytometry. The results showed each treatment group demonstrated significant increase in body mass and colon length, decrease in colon mass, colon mass index, unit colon mass index, and histopathological score(P<0.05, P<0.01), reduction of the expression of p-STAT3, STAT3, p-STAT6, and STAT6(P<0.05, P<0.01), rise of the expression of SOCS1 and SOCS3(P<0.05, P<0.01), decrease of Tfh9 and Tfh17 cells, and increase of Tfr cells(P<0.05, P<0.01) compared with the model group. These results indicated that SSP and the split EP and WP may alleviate ulcerative colitis by inhibiting the activation of STAT/SOCS signaling pathway and regulating the balance of Tfr/Tfh9/Tfh17 cells.

Changes and significance of T follicular regulatory cells/T follicular helper cells and related cytokines in peripheral blood of children with dust mite allergic asthma / 中华实用儿科临床杂志

Objective:To explore the changes of T follicular regulatory (T FR) cells/T follicular helper (T FH) cells and their related cytokines in peripheral blood of children with dust mite allergic asthma, and their clinical significance. Methods:A total of 25 children with acute dust mite allergic asthma (the asthma group) in Affiliated Hospital of Jiangnan University from January to December 2021 and 16 age- and sex-matched healthy volunteers (the healthy control group) at the same time were enrolled in the retrospective study.The percentages of peripheral T FR cells and T FH cells of the 2 groups were measured by flow cytometry.The plasma levels of cytokines[interleukin (IL)-10, IL-21] of the 2 groups were assessed by the flow cytometric microsphere-based array technology.The specific IgE (sIgE) levels of dust mites in 2 groups were detected by fluorescence enzyme immunoassay.The percentage of eosinophils in peripheral blood detected by blood cell analyzer.Data between groups were compared by t-test, and the correlation among indicators was analyzed by Spearman rank correlation analysis. Results:The asthma group had evident T FR cells/T FH cells immune imbalance.Compared with the healthy control group, the asthma group had a significantly lower T FR cells level[(0.11±0.03)% vs.(0.13±0.03)%], a significantly higher T FH cells level[(5.07±1.75)% vs.(3.80 ± 1.60)%], and a significantly lower ratio of T FR cells /T FH cells(0.02±0.01 vs.0.05±0.03) ( t=2.29, 2.30, 3.71; all P<0.05). Compared with the healthy control group, the asthma group had a significantly higher IL-21 level[(547.85±195.13) ng/L vs.(404.94±110.41) ng/L], and a significantly lower IL-10 level[(10.18±3.49) ng/L vs.(14.79±5.65) ng/L] ( t=2.60, 3.15; all P<0.05). The ratio of T FR cells/T FH cells in asthma group was negatively correlated with sIgE ( r=-0.444 2, P=0.026 1), but not related to the eosinophil percentage ( r=-0.135 2, P=0.519 3). Conclusions:Children with dust mite allergic asthma suffer from T FR cells/T FH cells subset imbalance.The imbalanced T FR cells, T FH cells and their related cytokines IL-10 and IL-21 may play a role in regulating the production of asthma sIgE.

Correlation between T follicular helper cells and eosinophilic gastroenteritis in children / 中华实用儿科临床杂志

Objective:To investigate the role and clinical significance of T follicular helper (Tfh) cells in the pathogenesis of eosinophilic gastroenteritis (EG) in children.Methods:A total of 17 children diagnosed with EG in the Department of Gastroenterology, Children′s Hospital of Nanjing medical University from October 2018 to January 2020 were recruited as EG group.During the same period, 15 children diagnosed with colon polyps were included as control group.Flow cytometry was used to detect Tfh cells and their functional molecules, including inducible costimulatory molecules (ICOS) and programmed cell death protein 1 (PD-1) in peripheral blood of the 2 groups.Results:The median (interquartile range) of Tfh cell frequency in peripheral blood of children with EG group was 7.3 (2.6)%, which was significantly higher than that of controls 2.8 (1.4)% ( P<0.05). There were significant differences in the median (interquartile range) of ICOS [1.5 (1.3)% vs.0.1 (0.2)%] and PD-1 expressions [1.8 (3.2)% vs.0.7 (0.6)%] on Tfh cells between children with EG group and control group (all P<0.05). The frequency of Tfh cells in the peripheral blood of children with EG was positively correlated with the expressions of ICOS ( r=0.746, P<0.05) and PD-1 ( r=0.893, P<0.05), and immunoglobulin E (IgE) level ( r=0.587, P<0.05). Conclusions:The frequency of Tfh cells in peripheral blood of children with EG significantly increases, which are proliferative and overexpressed with ICOS and PD-1.Moreover, the frequency of Tfh cells of EG patients is positively correlated with the level of IgE.The abnormal expression of Tfh cells may play a promoting role in the mechanism of EG.

Relationship between subtypes of T follicular helper cells and unexplained recurrent spontaneous abortion / 上海交通大学学报（医学版）

Objective · To discuss the relationship between unexplained recurrent spontaneous abortion (URSA) and T follicular helper (Tfh) cell subtypes. Methods · Twenty-eight normal early pregnancy women, who had undergone induced abortion, were taken as control, and 28 patients with URSA were enrolled in the abortion group. The mononuclear cells in peripheral blood and decidual tissues were separated in the two groups, and CXCR3+CCR6- Tfh cells, CXCR3-CCR6- Tfh cells, CXCR3-CCR6+ Tfh cells and B cells were tested by flow cytometry. Results · The decidual CXCR3-CCR6- Tfh cells significantly increased in the abortion group compared with control group (P=0.015). And there was a strong association between the decidual CXCR3-CCR6- Tfh cells and B cells in URSA patients (R2=0.779, P=0.025). Conclusion · The up-regulation of decidual CXCR3-CCR6- Tfh cells in early pregnancy women may be related with the occurrence of URSA.

Relationship between subtypes of T follicular helper cells and unexplained recurrent spontaneous abortion / 上海交通大学学报（医学版）

Objective · To discuss the relationship between unexplained recurrent spontaneous abortion (URSA) and T follicular helper (Tfh) cell subtypes. Methods · Twenty-eight normal early pregnancy women, who had undergone induced abortion, were taken as control, and 28 patients with URSA were enrolled in the abortion group. The mononuclear cells in peripheral blood and decidual tissues were separated in the two groups, and CXCR3+CCR6- Tfh cells, CXCR3-CCR6- Tfh cells, CXCR3-CCR6+ Tfh cells and B cells were tested by flow cytometry. Results · The decidual CXCR3-CCR6- Tfh cells significantly increased in the abortion group compared with control group (P=0.015). And there was a strong association between the decidual CXCR3-CCR6- Tfh cells and B cells in URSA patients (R2=0.779, P=0.025). Conclusion · The up-regulation of decidual CXCR3-CCR6- Tfh cells in early pregnancy women may be related with the occurrence of URSA.

Mechanism of T follicular cells in experimental autoimmune encephalomyelitis / 中国免疫学杂志

Objective: To evaluate the mechanism of T follicular helper cells ( Tfh ) in experimental autoimmune encephalomyelitis (EAE) via in vivo experiments. Methods:C57BL/6 mice were randomly divided into four groups,CFA group,EAE group,anti-ICOSL group and control group. Lymphocytes of different time points isolated from draining lymph nodes and spleen were stained for T follicular helper cells surface marker and T cells activation surface marker and analyzed by FACS. Observed parameters include inflammatory infiltration,demyelination in spinal cord and germinal center in spleen. ELISA was used to measure the level of antigen specific antibodies. Results: Mice in anti-ICOSL treated group developed mild disease was with lower clinical scores when compared with the EAE group. HE staining results turned out with alleviated inflammation and Luxol Fast Blue staining( LFB) showed no demyelization in anti-ICOSL treated mice compared with non-treated EAE models. Flow cytometry results revealed that percentages of T follicular helper cells decreased though the whole activated degree T cells was not influenced in anti-ICOSL treated group. Fewer ger minal center was found in both anti-ICOSL group and CFA group with reduced secretion of MOG-specific Ab. Conclusion:T follicular helper cells supported the development of cognate B cells,promoted the formation of germinal center,facilitate pathogenic MOG-specific Ab secretion,thus enhance EAE.

Relationships between the level of chemokine CXCL13 and the distribution of Tfh subsets in peripheral blood of hepatitis B patients and their clinical significance / 中华微生物学和免疫学杂志

Objective To investigate the distribution of T follicular helper(Tfh)cell subsets in hepa-titis B patients at different immune stages and to clarify the relationships between the level of CXCL13 and the distribution of Tfh cell subsets. Methods Flow cytometry analysis was performed to detect the distribution of Tfh cells. Enzyme-linked immunosorbent assay(ELISA)was performed to measure the levels of CXCL13 in ser-um samples collected from hepatitis B patients. RT-PCR and Western blot assay were used to analyze the expres-sion of CXCL13 in HepG2 and HepG2. 2. 1. 5 cells. Results The percentages of Tfh1 cells were significantly up-regulated at the immune activation(IA)stage,while those of Tfh2 cells were significantly raised at the im-mune tolerance(IT)stage. The percentages of Tfh17 cells in patients at the stage of IT were similar to those in patients at the stage of IA,but were higher than those in responders with HBsAg seroconversion(RP)or healthy controls(HC). The expression of CXCL13 was positively correlated with the percentage of Tfh2 cells. More over,hepatitis B virus(HBV)enhanced the expression of CXCL13 at both transcriptional and translational lev-els. Conclusion HBV might up-regulate the percentage of Tfh2 cells through promoting the expression of CXCL13,which resulted in the induction of immune tolerance. Elucidating the functions of Tfh1,Tfh2 and Tfh17 cells and understanding the type conversion mechanism among the three subsets are important for further researches on HBV-induced immunosuppression.

ClinicaI significance of T follicuLar helper cells in rheumatoid arthritis / 国际检验医学杂志

Objective To investigate the percentages of T follicuLar helper cells (Tfh)and interleukin-21(IL-21)in the plasma of patients with rheumatoid arthritis,and the immunological mechanism of Tfh cells in the development of rheumatoid arthritis. Methods According to ACR and DAS28,patients with rheumatoid arthritis were divided into low,moderate and high activity group.The percentages of CD3 + CD4 + CXCR+ 、CD3 + CD4 + ICOS+ and CD3 + CD4 + CXCR+ ICOS+ Tfh cells were detected by Flow Cytometry.While the levels of IL-21 、RF-IgM and anti-CCP in plasma were measured by ELlSA test.The analysis was performed by t-test and Spearman′s correlation analysis.Results The expression of CD3 + CD4 + CXCR+ ICOS+ Tfh cells in PBMCs of rheu-matoid arthritis was significantly higher than that in the normal controls(P <0.05).Meanwhile the results of the three rheumatoid arthritis groups(low,moderate and high activity groups)showed that the expression of Tfh increased accordingly(P <0.05).The expression of Tfh in rheumatoid arthritis was positively related with the levels of IL-21 ,ESR,CRP,RF and anti-CCP respectively. Conclusion The expression of Tfh and IL-21 increases significantly and is closely related to the disease activity in rheumatoid ar-thritis.The results indicate that the abnormality of Tfh may play an important role in the pathogenesis of rheumatoid arthritis.

Alteration of T follicular helper cells in children with persistent immune thrombocytopenic purpura / 中华微生物学和免疫学杂志

Objective To investigate the role of T follicular helper ( Tfh) cells in the immuno-pathogenesis of persistent immune thrombocytopenic purpura ( pITP) .Methods Twenty children with pITP and twenty healthy controls were enrolled in this study .The proportion of CD4+CXCR5+ICOShigh PD-1high T ( cTfh) cells and the expression of ICOSL on CD 19+B cells in peripheral blood of the patients and healthy subjects were analyzed by flow cytometry .The expressions of Bcl-6, c-Maf, IL-21 and ICOSL at mRNA level were detected by real-time PCR.The plasma concentrations of IL-2, IL-6 and IL-21 were determined by ELISA.Results (1) Compared with the healthy controls , the proportions of cTfh cells increased signifi-cantly in patients with pITP [(17.45±9.04) %vs.(7.57±2.57) %, P0.05).(4) The plasma concentration of IL-21 was remarkably elevated in patients with pITP , regardless of DEX treatment (P0.05).Con-clusion The immunopathogenesis of persistent immune thrombocytopenic purpura might be associated with the hyper-activation of Tfh cells caused by excessive expression of ICOSL and IL-21.The persistent high expression of ICOSL and IL-21 might be one of the important factors resulting in the recurrence of pITP in children .

The Current Researches on T Follicular Helper Cells and Associated Diseases.

T follicular helper (Tfh) cell is a new subpopulation of CD4+ T cell family, whose differentiation is affected by Bcl-6, Blimp-1, STAT3, STAT5 and so on, and it could affect or decide the development of other subsets of CD4+ T cells. The important function of Tfh cell is to help B cell mediate humoral immunity, many researches have proved that Tfh cells participate in the development of autoimmune disease, immunodeficient disease, tumor and infectious diseases.

Alteration mechanics of T follicular helper cell in Kawasaki disease / 中华微生物学和免疫学杂志

ObjectiveTo study the alteration mechanism of T follicular helper (Tfh) cells in patients with Kawasaki disease (KD).MethodsTwenty children with KD and the same number of agematched healthy subjects were studied.The proportion of CD4+CXCRS+ICOS+ T in peripheral blood was analyzed by flow cytometry.Real-time PCR was performed to detect the level of Tfh transcriptional factor( Bcl6) and its inhibitor( Blimp-1 ).The plasma concentration of IL-4 and IL-21 were determined by ELISA.ResultsThe proportion of CD4+CXCR5+ICOS+ T in patients with KD was significantly higher than healthy controls [ (2.6±0.6) ％ vs ( 1.8±0.7 ) ％,P＜0.05 ].Transcription levels of Bcl-6 were significantly elevated in patients with KD( P＜0.05),its inhibitor Blimp-1 were found to be down-regulated during acute phase of KD compared with healthy controls (P ＜ 0.05 ).The significant increase of IL-4 and IL-21 plasma concentrations were detected in patients with KD(P＜0.05),in comparison with healthy controls.ConclusionThe over activity of Tfh might be correlated with immune dysregulation in Kawasaki disease.Dysregulation of Bcl-6/Blimp-1,altered microenvironment of IL-4 and IL-21 might be correlated with the abnormal activity of Tfh cells.