Unbalanced expression of aryl hydrocarbon receptor in peripheral blood CCR6+CD4+ and CD4+CD25+T cells of rheumatoid arthritis / Expressão não equilibrada do receptor de hidrocarboneto arílico nos linfócitos T CCR6+ CD4+ e CD4+ CD25+ do sangue periférico na artrite reumatoide

ABSTRACT Objective: The goal of this study was to analyze the role of aryl hydrocarbon receptor in peripheral blood CCR6+CD4+ and CD4+CD25+T cells of patients with rheumatoid arthritis. Methods: Flow cytometry was applied to determine the proportion of AhR positive cells in CCR6+CD4+T, CD4+CD25+T and peripheral blood peripheral mononuclear cells from each subject. AhR mRNA and CYP1A1 mRNA relative expression levels were tested by real-time PCR. Results: The percentage of AhR positive cells in peripheral blood mononuclear cells was higher in RA group than that in healthy cases [(35.23 ± 10.71)% vs. (18.83 ± 7.32)%, p < 0.01]. The expression levels of AhR and CYP1A1 were both increased in patients with RA while compared to controls [(3.71 ± 1.63) vs. (2.00 ± 1.27), p = 0.002; (2.62 ± 2.08) vs. (0.62 ± 0.29), p < 0.01, respectively]. In RA patients, the percentage of AhR positive cells in CD4+CD25+T cells was significantly lower than that from controls [17.90 (6.10 ± 80.10)% vs. (52.49 ± 19.18)%, p < 0.01]; In healthy controls, the percentage of AhR positive cells in CD4+CD25+T cells was significantly higher than that in CCR6+CD4+T cells, and was also significantly higher than that in PBMCs [(52.49 ± 19.18)% vs. (23.18 ± 5.62)% vs. (18.06 ± 7.80)%, X 2 = 24.03, p < 0.01]; in RA patients, the percentage of AhR positive cells in CCR6+CD4+T cells was significantly increased than that in CD4+CD25+T cells and PBMCs [(46.02 ± 14.68)% vs. 17.90 (6.10 ± 80.10)% vs. (34.22 ± 10.33)%, X 2 = 38.29, p < 0.01]; Nevertheless, no statistically significant relationship was found between clinical data and AhR positive cells in CCR6+CD4+T and CD4+CD25+T cells. Conclusion: AhR may participate in the pathological progress of RA by controlling the differentiation of Th17 and Treg cells in peripheral blood.

RESUMO Objetivo: Analisar o papel do receptor de hidrocarboneto arílico (AhR) nos linfócitos T CCR6+ CD4+ e CD4+ CD25+ no sangue periférico de pacientes com artrite reumatoide (AR). Métodos: Foi aplicada citometria de fluxo para determinar a proporção de células AhR positivas em linfócitos CCR6+ CD4+ e CD4+ CD25+ do sangue periférico e células mononucleares periféricas de cada indivíduo. Os níveis de expressão relativa de ácido ribonucleico mensageiro (do inglês ribonucleic acid, RNAm,) de AhR e RNAm de enzima de primeiro estágio essencial para o AhR (CYP1A1) foram testados por reação em cadeia de polimerase (do inglês polymerase chain reaction, PCR,) em tempo real. Resultados: A percentagem de células AhR positivas nas células mononucleares do sangue periférico foi maior no grupo com AR do que nos indivíduos saudáveis [(35,23 ± 10,71)% vs. (18,83 ± 7,32)%, (p < 0,01)]. Os níveis de expressão de AhR e CYP1A1 estavam aumentados em pacientes com AR quando comparados com os controles [(3,71 ± 1,63) vs. (2,00 ± 1,27), p = 0,002; (2,62 ± 2,08) vs. (0,62 ± 0,29), p < 0,01, respectivamente]. Em pacientes com AR, a percentagem de células AhR positivas nos linfócitos T CD4+ CD25+ foi significativamente inferior à dos controles [17,90 (6,10 ± 80,10)]% vs. (52,49 ± 19,18)%, p < 0,01]; em controles saudáveis, a percentagem de células AhR positivas nos linfócitos T CD4+ CD25+ foi significativamente mais elevada do que nos linfócitos T CCR6+ CD4+ e também foi significativamente maior do que nas células mononucleares do sangue periférico (do inglês peripheral blood mononuclear cells, PBMC,) [(52,49 ± 19,18)% vs. (23,18 ± 5,62)% vs. (18,06 ± 7,80)%, X 2 = 24,03, p < 0,01]; em pacientes com AR, a percentagem de células AHR positivas nos linfócitos T CCR6+ CD4+ era significativamente maior em comparação com os linfócitos T CD4+ CD25+ e PBMC (46,02 ± 14,68)% vs. [17,90 (6,10 ± 80.10)]% vs. (34,22 ± 10,33)%, X2 = 38,29, p < 0,01]; no entanto, não foi encontrada correlação estatisticamente significativa entre os dados clínicos e células AhR positivas em linfócitos T CCR6+ CD4+ e CD4+ CD25+. Conclusão: O Ahr pode participar do progresso patológico da AR ao controlar a diferenciação de linfócitos Th17 e Treg no sangue periférico.

The effect of TGP assisted treatment of SLE on the expression of CD4+CD25+T in patient peripheral blood / 重庆医学

Objective to investigate the effect of TGP assisted treatment of SLE on the expression of CD4+CD25+ T in patient peripheral blood .Methods flow cytometry was used to detect the peripheral blood CD4+ CD25+ T cells in healthy group ,routine group and TGP group .Results The expression rate of CD4+CD25+ T cells in SLE patients was (6 .15 ± 1 .21)% ,and that of the healthy controls was (12 .30 ± 1 .78)% .The expression rate of CD4+CD25+ T cells in active SLE patients and healthy controls were significantly different (t=22 .03 ,P<0 .05) .In the routine group ,the expression rate of peripheral blood CD4+CD25+ T cells before and after treatment were significantly different (t= 12 .30 ,P<0 .05);in the TGP group ,the expression rate of peripheral blood CD4+ CD25+ T cells before and after treatment were significantly different ,too (t=16 .68 ,P<0 .05) .The expression rate of periph‐eral blood CD4+CD25+ T cells in routine group and TGP group were (9 .34 ± 1 .37)% and (11 .49 ± 1 .14)% respectively ,and the difference was statistically significant (t=6 .46 ,P<0 .05) .Conclusion The expression rate of CD4+ CD25+ T cells in active SLE patients was significantly lower than that of healthy controls ;the expression rate of CD4+CD25+ T cells in SLE patients increased significantly after treatment .The TGP treatment may work on CD4+CD25+ T cells .

Effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro / 中国免疫学杂志

Objective:To explore the promoting effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro and construct a stable culture system in vitro for CD 4+CD25+regulatory T cells from human umbilical cord blood.To compare the inhibiting effects between induced proliferated CD 4+CD25+Tregs and naturally isolated CD 4+CD25+Tregs on PBMCs functional activity.Methods:CD4+CD25-T cells and CD4+CD25+T cells were isolated from human umbilical cord blood mononuclear cells by magnetic activated cell sorting ( MACS) system and then expanded in vitro.Four different concentration levels of IL-7 combined with proper concentration of IL-2 were added as inducer and the efficiency and optimal concentration of IL-7 on inducing,CD4+CD25-T cells were analyzed via 4 different methods.Flow cytometry method was used to detect the changes of CD 4+CD25-T cells.The inhibitory effect of expanded CD 4+CD25+T cells on peripheral blood mononuclear cells (PBMCs) was tested by MTS.The expressions of Foxp3,IL-10 and TGF-βgenes in CD4+CD25+T cells were test by RT-PCR.Results:The CD4+CD25+T cells from each groups were expanded significantly after three weeks of culture.The results indicated that use of IL-7 combined with IL-2 resulted in the highest cell expansion comparing to the other groups.It was shown by the inhibitory test that the expanded CD 4+CD25+regulatory T cells could inhibit the proliferation of PBMCs ,but IL-7 induced CD4+CD25+regulatory T cells exerted weaker suppressor activity than natural regulatory T cells .Only IL-7 (4 ng/ml) and IL-2 (2 000 U/ml) induced CD4+CD25+regulatory T cells showed the strongest killing activity.Conclusion:We successfully expand CD4+CD25+regulatory T cells in vitro.The protocol is established in which the use of mAbCD 3/CD28 combined with IL-7 and IL-2 resulted in the highest cell expansion ,and intensely expressed cell phenotype of CD 4 and CD25.

Isolation, identification and functional characteristics of murine B10 cells / 中华微生物学和免疫学杂志

Objective To investigate the phenotypes and percentages of B 10 cells in different tis-sues from wild-type mice and to identify their biological functions .Methods The percentages of B10 cells derived from different tissues of mice and their responses to lipopolysaccharide ( LPS) stimulation were ana-lyzed by flow cytometry .Magnetic-activated cell sorting ( MACS ) and fluorescence-activated cell sorting (FACS) were used to purify B10 cells, CD4+CD25-T cells and Treg cells.CD4+CD25-T cells and Treg cells labeled by CFSE were co-cultured with or without B10 cells, and then their proliferation were evaluated after 72 h.Results (1) A subset of CD19+CD5+CD1dhigh regulatory B cells was identified in spleen , pe-ripheral blood and lymph nodes from wild-type mice , of which the highest frequency was detected in spleen (3.95%±0.79%, P<0.05).The isolated B cells from different tissues were stimulated by LPS , PMA, ionomycin and monensin (L+PIM) in vitro to express IL-10.Among them, splenic CD19+IL-10+B cells showed the highest expression of IL-10 (P<0.05).(2) Prolonged LPS stimulation (48 h) to CD5+CD1dhigh B cells enhanced the expressions of IL-10 (P<0.01).(3) CD19+CD5+CD1dhigh B cells inhibited the prolif-eration of CD4+CD25-T cells in vitro in a dose-dependent manner (P<0.01), but increased the secretion of IL-10 by CD4+T cells (P<0.01) and the proliferation of Treg cells in vitro (P<0.01).Conclusion Com-pared with other tissues , the percentage of B10 cell subset in spleen is the highest in wild-type mouse , and B10 cells subset can be activated through Toll-like receptor ( TLR ) signaling pathway .The responses of CD4+CD25-T cells and Treg cells in co-culture with B10 cells are regulated by B 10 cell subset through an increased IL-10 production .B10 cells might be a useful cell population for the treatment of inflammatory au-toimmune diseases.

Role of glucagon-like peptide-1 analogue liraglutide played in the proliferation of CD4+ CD25-T cells in normal people and type 1 diabetic patients in vitro / 中华内分泌代谢杂志

Objective To study the role of glucagon-like peptide-1 (GLP-1) analogue liraglutide played in the proliferation of CD4+CD25 T cells in normal people and newly-onset type 1 diabetic patients,and to evaluate the possible immune regulatory role of liraglutide in the therapy of type 1 diabetes.Methods CD4+ CD25-T cells of 10 normal people and 10 newly-onset type 1 diabetic patients were separated from peripheral blood by MACS immunomagnetic beads and stimulated by Human T-Activator CD3/CD28 Dynabeads to proliferate.CFSE labeling technique was used to evaluate the proliferation of CD4+ CD25-T cells by flow cytometry.Liraglutide of different concentrations(0,25,50,and 100 nmol/ml) was added to the proliferation system,then the proliferation of CD4+CD25-T cell was measured.Results (1) Liraglutide suppressed the proliferation of CD4+ CD25-T cells from either normal people or type 1 diahetic patients with dose-dependent manner (P ＜ 0.05).(2) Under the different concentrationsofliraglutide,the proliferation ofCD4+CD25 T cells from diabetic patients was mueh more robust than that of normal people (P＜0.01).(3) The inhibitory effects of liraglutide on CD4+ CD25-T cells proliferation in normal people and diabetic patients were similar (P＞0.05).Conclusion The proliferation of CD4+ CD25 T cells in type 1 diabetic patients was more robust than normal people,which indicated cellular immune dysfunction in type 1diabetes.Liraglutide inhibits the proliferation of CD4+ CD25-T cells of type 1 diabetic patients in vitro.The immunosuppression effect of liraglutide may have potential value in the treatment of type 1 diabetes.

Indoleamine 2,3-dioxygenase and immune tolerance in tumor / 国际肿瘤学杂志

Clonal suppression induced by CD4+ CD25+ regulatory T cells is one of the principal factors to evoke the immune tolerance in tumor. Through the activation of CD4+ CD25+ regulatory T cells, the indoleamine 2,3-dioxygenase (IDO) reduces the immune response in tumor micro-environment of various systems , and induces the formation of host immune tolerance. IDO inhibitor 1-MT is expected to become a new target for cancer treatment.

The inhibitory effect of gadolinium chloride on the regulatory T cells of mice with hepatic granuloma caused by Schistosoma japonicum / 中国人兽共患病学报

To determine whether the targeting inhibition of Kupffer cellsfunction mediated by gadolinium chloride (GdCl_3) could interfere with the CD4~+CD25~+ regulatory T cells of mice at the granuloma stage of schistsomiasis, female C57BL/6 mice of 6-8 weeks old were divided randomly into 3 groups, i.e. control group. group infected with cercariae of Schistsoma japonicum and group of infection plus GdCl3,. GdCl3 in a dosage of 15 mg/kg was introduced into mice through penile vein twice per week. The number of CD4~+CD25~+ T cells was determined using flow cytometry and the number of cells with Fox p3 was detected by using immunohistochemical methods. For detection of cytokines, such as IL-4, IL-5, IL-10, TGF-β1, ,IFN-γ in mouse sera, a DuoSer ELISA development kit was used, It was found that the number of CD4~+CD25~+ T cells and level of IL-10 in Schistosomiasis granuloma stage were decreased in the S.japonicum cercariae infected mice injected with GdCl_3 in comparison with the infection group. The percentages of CD4~+CD25~+ T cells of infection group and infection plus GdCl_3 group were 13.8%, 9.3% and 6.4% respectively, while the levels of IL-10 of these 3 groups of rats were 41.4 pg/mL, 22.6 pg/mL and 11.5% respectively. In addition, treatment with GdCl_3 could down-regulate the expression of Fox p3 and reduce the inflammatory reactions in Schistosomiasis granuloma. It is evident that the targeting inhibition of Kupffer cellsfunction mediated by GdCI_3 interfere with the production of the regulatory T cells and reduce the inflammatory responses in Schistsomiasis granuloma.

The Binding Properties of Glycosylated and Non-Glycosylated Tim-3 Molecules on CD4+CD25+ T Cells

BACKGROUND: T cell immunoglobulin and mucin domain containing 3 protein (Tim-3) expressed on terminally differentiated Th1 cells plays a suppressive role in Th1-mediated immune responses. Recently, it has been shown that N-glycosylation affects the binding activity of the Tim-3-Ig fusion protein to its ligand, galectin-9, but the binding properties of non-glycosylated Tim-3 on CD4+CD25+ T cells has not been fully examined. In this study, we produced recombinant Tim-3-Ig fusion proteins in different cellular sources and its N-glycosylation mutant forms to evaluate their binding activities to CD4+CD25+ T cells. METHODS: We isolated and cloned Tim-3 cDNA from BALB/C mouse splenocytes. Then, we constructed a mammalian expression vector and a prokaryotic expression vector for the Tim-3-Ig fusion protein. Using a site directed mutagenesis method, plasmid vectors for Tim-3-Ig N-glycosylation mutant expression were produced. The recombinant protein was purified by protein A sepharose column chromatography. The binding activity of Tim-3-Ig fusion protein to CD4+CD25+ T cells was analyzed using flow cytometry. RESULTS: We found that the nonglycosylated Tim-3-Ig fusion proteins expressed in bacteria bound to CD4+CD25+ T cells similarly to the glycosylated Tim-3-Ig protein produced in CHO cells. Further, three N-glycosylation mutant forms (N53Q, N100Q, N53/100Q) of Tim-3-Ig showed similar binding activities to those of wild type glycosylated Tim-3-Ig. CONCLUSION: Our results suggest that N-glycosylation of Tim-3 may not affect its binding activity to ligands expressed on CD4+CD25+ T cells.

Rosiglitazone upregulates Foxp3 mRNA expression of CD4+T cells in adults with latent autoimmune diabetes / 中华微生物学和免疫学杂志

Objective To investigate the effect of rosiglitazone on the CD4+regulatory T cells in the patients with latent autoimmune diabetes in adults(LADA).Methods The CIM+T cells from IADA patients were isolated with anti-CD4-dynal magnetic beads.The expression of Foxp3 mRNA,along with peroxisome proliferators activator receptors gamma(PPARγ)mRNA and TGF-131 mRNA was determined.The effect of rosiglitazone on CD4+T cells was measured,after treated with rosiglitazone for 48 h.Cell viability was assessed by Mtit assay.The proliferation was assayed with 3 H-TdR.Two-color staining(anti-CD4,anti-CD25)flow cytometric analysis was employed to measure the percentage of CD4+CD25+T cells of Deriph eral blood.Resuits PPARγmRNA was expressed in peripheral CD4+T lymphocytes.RosiglitazoBe inhibited phytohemagglutinin(PHA)-induced human CD4+T cell proliferation in dose dependence.The percentage of CD4+CD25+T cells showed no significant change after the peripheral blood culture with 1 μmol/Land 10μmot/L rosiglitazone.10 μmol/L of rosiglitazone induced Foxp3 mRNA expression in vitro (3.27fold,P<0.05),whereas TGF-β1 mRNA expression did not change.Furthermore,only 1 μmol/L of rosiglitazone could promote Foxp3 mRNA expression if adding IL-2(10 U/m1)in cultures(3.48 fold.P<O.05).Conclusion Rosiglitazone promotes Foxp3 mRNA expression in CD4+T cells in vivo and in vitro,improving the defective of the regulatory T cells.

Regulatory T Cells and Allogeneic Transplantation / 소아과

Allogeneic organ or hematopoietic stem cell transplantation(HSCT) is the treatment of choice for end-stage organ diseases or various hematologic disorders. The induction of alloantigen specific T cell tolerance and its maintenance are critical for preventing immune responses, including graft rejection or graft-versus-host disease(GVHD) in allogeneic transplantation. CD4+ T cells are classified as immune functions : Th1 CD4+ cells for cellular immunity, Th2 for humoral immunity, Th3 for suppressive effect against activated T cells, Tr1 for regulation of immune response. Some CD4+ regulatory T cells have a major role in controlling immune response to alloantigen. A minor population of CD4+ T cells, which co-express the interleukin-2 receptor(IL-2R) alpha-chain(CD25), is crucial for the control of autoreactive T cells and for peripheral tolerance in allogeneic transplantation. CD4+CD25+ regulatory T cells express high level of cytotoxic T lymphocyte associated antigen 4(CTLA 4), as cell-contact mechanism, and secret immunomodulating cytokines, as IL-10 or TGF-beta, as independent cell contact. High expression of CTLA 4 on CD4+CD25+ T cells may contribute to deliver suppressive signals into T cells via CD28. Immature dendritic cells have low expression of major histocompatibility(MHC) and co-stimulatory signals, and few secretion of IL-12. CD4+CD25+ T cells are developed by immature myeloid dendritic cells, which are controlled under vitamin D3 or IL-10. In kidney transplantation, graft survival in recipients with donor specific transfusion(DST) showed longer than without DST. DST may induce antigen specific CD4+CD25+ T cells, and these cells play a role for central or peripheral tolerance against immune cells. In allogeneic HSCT, donor CD4+CD25+ T cells suppress lethal GVHD in MHC mismatched pairs, and also may possess graft-versus-leukemia effect in early infusion with HSC. We need more study for cytotoxic effect of CD4+CD25+ T cells, which have Fas-FasL interaction, although these cells in cancer patients suppress autoreactive T cells against tumor. Recently, many trials have investigated treatment for intractable disease using various types of cells, including mesenchymal stem cells, dendritic cells. We have to consider ex vivo expansion of CD4+CD25+ T cells for induction of immune tolerance in allogeneic transplantation. Now, we have to study or understand immunoregulatory cells for allogeneic transplantation or immune control.

Inhibition of Graft Versus Host Disease Using CD4+ CD25+ T Cells Induced with Interleukin-2 in Mismatched Allogeneic Murine Hematopoietic Stem Cell Transplantation

BACKGROUND: In kidney transplantation, donor specific transfusion may induce tolerance as a result of some immune regulatory cells against the graft. In organ transplantation, the immune state arises from a relationship between the immunocompromised graft and the immunocompetent host. However, a reverse immunological situation exists between the graft and the host in hematopoietic stem cell transplantation (HSCT). In addition, early IL-2 injections after an allogeneic murine HSCT have been shown to prevent lethal graft versus host disease (GVHD) due to CD4+ cells. We investigated the induction of the regulatory CD4+CD25+ cells after a transfusion of irradiated recipient cells with IL-2 into a donor. METHODS: The splenocytes (SP) were obtained from 6 week-old BALB/c mice (H-2(d)) and irradiated as a single cell suspension. The donor mice (C3H/ He, H-2(k)) received 5x10(6) irradiated SP, and 5,000 IU IL-2 injected intraperitoneally on the day prior to HSCT. The CD4+CD25+ cell populations in SP treated C3H/He were analyzed. In order to determine the in vivo effect of CD4+CD25+ cells, the lethally irradiated BALB/c were transplanted with 1x10(7) donor BM and5x10(6) CD4+CD25+ cells. The other recipient mice received either 1x10(7) donor BM with 5x10(6) CD4+ CD25- cells or the untreated SP. The survival and GVHD was assessed daily by a clinical scoring system. RESULTS: In the MLR assay, BALB/c SP was used as a stimulator with C3H/He SP, as a responder, with or without treatment. The inhibition of proliferation was 30.0 13% compared to the control. In addition, the MLR with either the CD4+CD25+ or CD4+CD25- cells, which were isolated by MidiMacs, from the C3H/He SP treated with the recipient SP and IL-2 was evaluated. The donor SP treated with the recipient cells and IL-2 contained more CD4+CD25+ cells (5.4+/-1.5%) than the untreated mice SP (1.4+/-0.3%)(P<0.01). There was a profound inhibition in the CD4+CD25+ cells (61.1+/-6.1%), but a marked proliferation in the CD4+CD25- cells (129.8+/-65.2%). Mice in the CD4+CD25+ group showed low GVHD scores and a slow progression from the post-HSCT day 4 to day 9, but those in the control and CD4+CD25- groups had a high score and rapid progression (P<0.001). The probability of survival was 83.3% in the CD4+CD25+ group until post-HSC day 35 and all mice in the control and CD4+CD25- groups died on post-HSCT day 8 or 9 (P=0.0105). CONCLUSION: Donor graft engineering with irradiated recipient SP and IL-2 (recipient specific transfusion) can induce abundant regulatory CD4+CD25+ cells to prevent GVHD.

Study on the blood CD4~+CD25~+T cells of rats with experimental autoimmune encephalomyelitis / 临床神经病学杂志

Objective To study the number changes of blood CD4+CD25+T cells and its significance in rats with experimental autoimmune encephalomyelitis (EAE). Methods Wistar rats were induced by guinea pig spinal cord homogenate (GPSCH) to establish EAE models.The number of peripheral blood CD4+CD25+T cells in EAE was detected by flow cytometry and compared with normal rats.The behavior and the pathological changes of brain and spine cord of rats were observed to ascertain EAE model successful.Results The achievement ratio of EAE model was 48.9%.The number of peripheral blood CD4+CD25+T cells of EAE rats( 5.29?4.00) was significantly lower than that in the control group( 12.61?2.24, P

Ovalbumin immune tolerance factors induce OVA specific peripheral immune tolerance in naive BALB/c mice / 中国免疫学杂志

Objective:To investigate the immunoregulation effect of ovalbumin immune tolerance factors (OVA ITFs).Methods:Components that were smaller than 3 kD were isolated from the splenic lymphocytes lysates of OVA tolerance mice or naive mice,respectively,named as OVA ITFs or OVA ITFs conttrol.Naive BALB/c mice were divided into 5 groups,group A,B,C,D were injected i.v.by OVA ITFs,OVA ITFs control,splenic lymphocytes from OVA tolerant mice or PBS,respectively,group E as the blank control.The percentages of CD4+CD25+ T cell subpopulation from spleens before and after adoptive transfer were measured with flow cytometer;OVA specific lymphocyte responses were assessed by MTT assay.The levels of IL-10 and TGF-?1 in the culture supernatants were tested by ELISA kits.Results:For OVA ITFs or splenic lymphocytes from OVA tolerant mice,the percentages of CD4+CD25+ T cell subpopulation from spleens after adoptive transfer were raised significantly compared with that before adoptive transfer (15.32%?1.03% and 15.35%?0.62% vs 9.97%?1.38%,P

The effect of cyclosporin A on CD4~+CD25~+T cells in TCR-transgenic DO11.10 mice immunized with OVA / 中国免疫学杂志

Objective:To study the effect of CsA on CD4+CD25+T cells from DO11.10 mice after immunization with OVA.Methods:The amount of spleen CD4+CD25+T cells and Foxp3+T cells was detected by means of flow cytometry (FCM).Results:After immunization with OVA,the ratios of CD4+CD25+T cells to CD4+T cells and Foxp3+T cells to CD4+CD25+T cells elevated in spleen,but CsA significantly decreased the percentage of CD4+CD25+T cells and CD4+CD25+Foxp3+T cells under either naive state or immune reaction state.Conclusion:CsA inhibits immune responses,while it also suppresses immune tolerance.

Effects of glucocorticoid on the expression of CD80 and CD4~+CD25~+T cells in peripheral blood lymphocyte of patients with multiple sclerosis / 临床神经病学杂志

Objective To investigate the effects of glucocorticoid(GC) on the expression of CD80 and CD4+CD25+T cells in peripheral blood lymphocyte of patients with multiple sclerosis (MS). Methods With flow cytometer,the positive rates of CD80 and CD4+CD25+T cells in peripheral blood lymphocyte were detected in 21 acute MS patients pre and post treatment separately,and compared with the nomal control group.The secols of Expanded Disablility Status Scale(EDSS) were compared pre and post treatment in MS patients.Results The positive rate of CD80[(5.031?1.782)%]in the MS patients pre treatment was obviously less than that in control group's [(6.436?2.035)%](P

CD_(25) Expression in Peripheral Blood T Lymphocytes from Patients with Chronic Hepatitis B and Its Significance / 中华医院感染学杂志

OBJECTIVE To investigate CD25 expression in peripheral blood T lymphocytes from patients with chronic hepatitis B,and explore its significance in the pathogenesis of chronic hepatitis B.METHODS To detect CD25 expression in peripheral blood T lymphocytes of patients with chronic hepatitis B by means of flow cytometry.CD25 expression was observed in chronic hepatitis B patients.In the meantime,CD25 expression in T cells from severe chronic hepatitis or acute hepatitis B patients and asymptomatic carriers of HBV was also observed.RESULTS Varying degrees of CD25 were expressed in T cells from hepatitis B patients.The expression in CD3+ T and CD8+T cells was higher than that in CD4+T cells.CD25 expression in CD4+T was lower.The average of CD25 expression in CD3+T cells from patients with chronic hepatitis B,acute hepatitis B,and chronic severe hepatitis B and asymptomatic carriers of HBV was(2.92?0.13)%,(0.51?0.36)%,(1.60?0.07)%,and(0.95?0.23)%,respectively.The average of CD25 expression in CD4+T cells from patients with chronic hepatitis B,acute hepatitis B and chronic severe hepatitis B and asymptomatic carriers of HBV was(2.58?0.50)%,(0.34?0.07)%,(1.45?0.02)%,and(0.83?0.13)%,respectively.CD3+T and CD4+T CD25 expression in patients with chronic hepatitis and,severe chronic hepatitis B was increased compared with that of acute hepatitis B patients and asymptomatic carriers of HBV.Compared with chronic severe hepatitis B,the expression of chronic hepatitis B was higher.CONCLUSIONS CD4+ CD25+T cells in chronic hepatitis B virus infection are increased compared with acute hepatitis,CD4+ CD25+T cells may be related to immune tolerance.

Mechanisms underlying the induction of IL-2 secretion by PDB plus ionomycin in CD4~+CD25~+ T cells from cord blood and adult peripheral blood / 中国病理生理杂志

AIM: To confirm that CD4~+CD25~+ regulatory T cells don't have an instinctive defection in IL-2 secretion, and to have an insight into the maturation state of CD4~+CD25~+ T cells in cord blood. METHODS: CD4~+CD25~+ and CD4~+CD25~- T cells were purified from cord blood of term infants (CB) and adult peripheral blood (PB) by autoMACS, and stimulated with PDB plus ionomycin. After 45 hours of culture, cells were detected for expression of CD69 and CD25 by flow cytometry, and the supernatants were measured for 7 kinds of cytokines by Luminex. RESULTS: CD4~+CD25~+ T cells from both CB and PB proliferated comparably with CD4~+CD25~- T cells when stimulated with PDB plus ionomycin. After 45 hours of culture, however, the CD4~+CD25~+ T cells underwent a tendency of cell death. Expression of CD25 was further upregulated when CD25~+ cells were activated. Under stimulation of PDB plus ionomycin, both CD4~+CD25~+ and CD4~+CD25~- T cells in PB secreted high levels of IFN-?, IL-2 and TNF-?, with CD25~+ cells secreted much higher level of IL-5, IL-4 and IL-10 than those in CD25~- cells; CD4~+CD25~+ and CD4~+CD25~- T cells in CB also secreted high level of IL-2 and TNF-? but much lower level of IFN-? than those in PB, and no secretion of IL-5, IL-4 and IL-10 was observed. CONCLUSION: CD4~+CD25~+ regulatory T cells don't have an instinctive defection in IL-2 secretion, otherwise there may be a different TCR signaling pattern in CD4~+CD25~+ T cells from traditional T cells. The CD4~+CD25~+ T cells in cord blood have not fully matured in function.