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Animal study on CD4^(+) CD25^(+) regulatory t cells for treating female mouse with recurrent spontaneous abortion

Discussion of Clinical Cases 2015年第4期2卷 1-5页

作者： Biao Duan Lu Wang Weiwei Huang Shouhong Wang Haiyan Du Reproductive Center of Department of Obstetrics and Gynecology The Third Affiliated Hospital of Inner Mongolia Medical UniversityBaoTouInner MongoliaChina

Objective:to explore immunotherapy effectiveness of the CD4^(+)CD25^(+) regulatory t cells for treating female mouse with recurrent spontaneous abortion(RSA)by animal experiments.Methods:Mononuclear lymphocytes were isolated from the blood(instead of cord blood)of new-born baby of KunMing Bai mouse or BALB/c male mouse with normal birth ability(as unrelated third party blood source)by density gradient centrifuga-tion method.the CD4^(+)CD25^(+) regulatory t cells were selected by magnetic-activated cell sorting from mononuclear cells of cord blood cells.CBA/J female mouse copulated with DBA/2J male mouse was utilized as RSA animal model.Pregnant RSA mice were injected different types of lymphocytes through tail vein.Independent sample t-test was used to analyze the data from each group.Results:the proportion of CD4^(+)CD25^(+)t cells in CD4^(+)t cells was(17.49±0.60)%in CD4^(+)CD25^(+) regulatory t cells injection group,which was statistical significant higher than that of mononuclear lymphocyte injection group(14.68±0.83)%,sterile PBS group(9.54±0.85)%or no injection group(9.28±0.68)%(pt-value was 4.754,13.242 and 15.621,respec-tively).the Foxp3 relative protein expression level of CD4^(+)CD25^(+) regulatory t cells injected group was 5.85±0.45,which was also significant higher than that of mononuclear lymphocyte injection(2.86±0.54),sterile PBS group(1.08±0.16)or no injection group(1.00±0.00)(pt-value was 7.276,17.227 and 18.635,respectively).Finally,two times of CD4^(+)CD25^(+)t cell injected group at the 4 th and 8 th day had well effect for RSA mouse,and embryo sorption rate was(4.92±0.08)%,which significant lower than that of two times of mononuclear lymphocyte injected group(13.07±0.06)%,sterile PBS group(23.11±0.12)%,or no injection group(25.47±0.11)%(pt-value was-2.603,-4.012 and-4.700,respectively).Conclusions:Pregnant mouse with RSA injected CD4^(+)CD25^(+)t cells several times for immunotherapy can get better effec-tiveness than that of pregnant mouse injected traditional mononuclear cel

关键词： regulatory t cells Recurrent spontaneous abortion Immune therapy Immune tolerance

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CD4^(+)CD25^(+)but not CD4^(+)Foxp3^(+) t cells as a regulatory subset in primary biliary cirrhosis

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Cellular & Molecular Immunology 2010年第6期7卷 485-490页

作者： Dandan Wang Huayong Zhang Jun Liang Zhifeng Gu Qiang Zhou Xiangshan Fan Yayi Hou Lingyun Sun Department of Immunology and Rheumatology The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjingChina Department of Pathology The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjingChina Immunology Laboratory Nanjing University Medical SchoolNanjingJiangsuChina

Increasing evidence indicates a role for regulatory t cells(tregs)in the immune response and in autoimmune diseases,but the role of tregs and cytokines in autoimmune hepatic diseases remains largely unclear and controversial,especially in patients with primary biliary cirrhosis(PBC).this study was undertaken to investigate tregs and different cytokines in the liver and peripheral blood of PBC patients.We found that these patients demonstrated a reduction of CD4^(+)CD25^(+) t cells but elevated CD4^(+)Foxp3^(+) t cells in peripheral blood mononuclear cells(PBMCs)and CD41 t cells.the percentage of CD4^(+)CD25^(+) t cells in PBMCs was negatively correlated with elevated plasma interferon(IFN)-c levels.A liver-specific analysis showed that the frequency of Foxp31 tregs,transforming growth factor(tGF)-b1 and IFN-c were increased in PBC patients.Our findings suggest that an imbalance between CD4^(+)CD25^(+) tregs and cytotoxic cytokines plays a crucial role in the pathogenesis of PBC while the role of Foxp3 needs further investigation.

关键词： cytokines forkhead box P3 primary biliary cirrhosis regulatory t cells

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regulatory t cells protect the liver from drug-induced immune-mediated injury

Regulatory T cells protect the liver from drug-induced immun...

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第八届全国免疫学学术大会

作者： Xuefu Wang Rui Sun Zhigang tian Institute of immunology School of Life Sciences University of Science and Technology of China

Background and Aim:Immune response is regarded to mediate the liver injury after initial response induced by overdose acetaminophen(APAP).However, the role of regulatory t(treg) cells in the following immune response is still poorly understood.Methods:the liver injury was assessed by alanine aminotransferase level and tissue staining.the levels of cytokines were measured by enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction.the lymphocyte populations were analyzed by flow cytometry.Results:Herein,we demonstrated that treg cells ameliorated the production of pro-inflammatory cytokines and attenuated the immune-exaggerated liver injury.the pro-inflammatory cytokines increased, accompa-nied with increasing CD44hiCD62L-CD4+t cells in the liver and enhanced liver injury after APAP treatment.Depletion of CD4+t cells obviously reduced the production of pro-inflammatory cytokines and ameliorated liver injury.Importantly, the percentage and absolute number of treg cells was reduced after APAP treatment.However, depletion of treg cells increased pro-inflammatory cytokines and exacerbated liver injury.Whereas, the adaptive transfer of treg cells reduced pro-inflammatory cytokines and the ALt levels.Conclusion:All of the results showed that the pro-inflammatory cytokines produced by th1 cells mediating APAP-induced liver injury were negatively regulated by treg cells.Our research suggests that hepatic treg cells maintain a local suppressive microenvironment that prevents drug-induced immune-exacerbated liver injury.the tolerance mechanism provides pivotal thoughts in the prevention and treatment of drug induced liver injury.

关键词： acetaminophen regulatory t cells Interferon-γ

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regulatory t cells及其表面标志物FoxP3在AA大鼠肺功能降低中的表达

Regulatory T cells及其表面标志物FoxP3在AA大鼠肺功能降低中的...

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中华中医药学会第十六届全国风湿病学术大会

作者：万磊刘健刘磊冯云霞程园园黄传兵汪元湖北中医药大学安徽中医学院第一附属医院安徽中医学院

目的:探讨调节性t细胞(treg)和其表面标志物叉状头转录因子(FoxP3)在RA肺病变中的发生机制。方法:采用弗氏完全佐剂复制佐剂关节炎模型(AA),用动物肺功能仪检测大鼠肺功能,采用流式细胞术检测大鼠外周血CD4+CD25+Foxp3+treg的表达,并采... 详细信息

目的:探讨调节性t细胞(treg)和其表面标志物叉状头转录因子(FoxP3)在RA肺病变中的发生机制。方法:采用弗氏完全佐剂复制佐剂关节炎模型(AA),用动物肺功能仪检测大鼠肺功能,采用流式细胞术检测大鼠外周血CD4+CD25+Foxp3+treg的表达,并采用Rt-PCR、免疫组化、免疫印迹检测大鼠肺组织Foxp3 mRNA和Foxp3 protein表达。结果:与正常组大鼠比较,AA大鼠组织肿胀度和关节炎指数升高,AA大鼠肺功能参数降低,外周血CD4+CD25+treg、CD4+CD25+FoxP3+treg表达降低,肺组织FoxP3 mRNA和Foxp3 protein表达水平亦降低(Ptreg表达降低,treg调控作用减弱,导致免疫平衡失调,免疫复合物在关节和肺组织沉积,进一步加重炎症反应,导致关节和肺组织损害,最终出现肺功能降低。

关键词： regulatory t cells FoxP3 Adjuvant arthritis Pulmonary function

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regulatory t cells及其表面标志物FoxP3在AA大鼠肺功能降低中的表达

Regulatory T cells及其表面标志物FoxP3在AA大鼠肺功能降低中的...

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全国第十届中西医结合风湿病学术会议

作者：万磊刘健刘磊冯云霞程园园黄传兵汪元湖北中医药大学安徽中医学院第一附属医院风湿免疫科安徽中医学院

目的探讨调节性t细胞(treg)和其表面标志物叉状头转录因子(FoxP3)在RA肺病变中的发生机制。方法采用弗氏完全佐剂复制佐剂关节炎模型(AA),用动物肺功能仪检测大鼠肺功能,采用流式细胞术检测大鼠外周血CD4CD25Foxp3treg的表达,并采用Rt-... 详细信息

目的探讨调节性t细胞(treg)和其表面标志物叉状头转录因子(FoxP3)在RA肺病变中的发生机制。方法采用弗氏完全佐剂复制佐剂关节炎模型(AA),用动物肺功能仪检测大鼠肺功能,采用流式细胞术检测大鼠外周血CD4CD25Foxp3treg的表达,并采用Rt-PCR、免疫组化、免疫印迹检测大鼠肺组织Foxp3 mRNA和Foxp3 protein表达。结果与正常组大鼠比较,AA大鼠组织肿胀度和关节炎指数升高,AA大鼠肺功能参数降低,外周血CD4CD25teg、CD4CD25FoxPtreg表达降低,肺组织FoxP3mRNA和Foxp3 protein表达水平亦降低(Ptreg表达降低,treg调控作用减弱,导致免疫平衡失调,免疫复合物在关节和肺组织沉积,进一步加重炎症反应,导致关节和肺组织损害,最终出现肺功能降低。

关键词： regulatory t cells FoxP3 Adjuvant arthritis Pulmonary function

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regulatory t cells Suppress Pathogenic CD8(+)t cells in Primary Biliary Cirrhosis Mice Model

Regulatory T cells Suppress Pathogenic CD8(+)T cells in Prim...

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第九届全国免疫学学术大会

作者：王寅虎杨微廉哲雄中国科学技术大学

Primary Biliary Cirrhosis(PBC)is a female dominant progressive autoimmune liver disease characterized by portal inflammation and immune-mediated destruction of intrahepatic bile ducts.Mice with t cell-restricted expression of a dominant negative form of transforming growth factorβreceptor typeⅡ(dntβRⅡ)spontaneously developed periductular aggregates in the liver.the precise relationship between the pathogenesis of PBC and regulatory t cells is poorly defined.Based on dntpRII mice,it was determined that the frequency of tregs didn’t change in lymph organs from dntpRII;Foxp3mice,compared with wild type(Foxp3)mice.CD4Foxp3tregs from dntβRⅡmice up-regulated suppressive functional molecule,such as CD25,CtLA-4,GItR,ICOS,CD69,and highly expressed th1 response or liver inflammation specific chemokine receptor CXCR3,meanwhile down-regulated CD62L(L-selectin),showed extremely activated phenotype.Besides,lacking of CD4t cells in dntβRⅡ:CD4mice aggravated PBC-like symptom compared with dntpRII mice.this data showed that in inflammatory milieu,regulatory t cells were over-activated.In the absence of CD4t cells,without the assistant from conventional CD4t cells and suppression from regulatory t cells,pathogenic t cells caused more severe liver inflammation.Moreover,Wt,dntpRII mice derived splenic tregs and in vitro tGF-P induced tregs(ti-tregs)showed obvious suppressive activity toward CD8t cells from dntβRⅡmice.Our research provided evidence for the protective role of regulatory t cells in this PBC mice model.After parabioted with wild type mice,dntβPⅡ:CD4mice showed almost no portal inflammation in liver and showed alleviated biliary disease.

关键词： Primary Biliary Cirrhosis regulatory t cells CD8 t cells Bone marrow transplantation Parabiosis

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IL-22 expression is correlated with FOXP3+ regulatory t cells infiltration and poor prognosis in patients with breast cancer

IL-22 expression is correlated with FOXP3+ regulatory T cell...

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第十三届全国免疫学学术大会

作者： Xiangmei Zhang Baoen Shan Yunjiang Liu Fourth Hospital of Hebei Medical University

Purpose: Interleukin-22(IL-22) is involved in the modulation of tissue responses during inflammation and the interaction between IL-22 production and FOXP3 regulatory t cells(tregs) for maintain the immune homeostasis has been identified. However, little is known about the role of IL-22 in breast cancer and the association between IL-22 and FOXP3 expression.Methods: tissues were obtained from 152 cases of breast invasive ductal carcinoma and IL-22 and FOXP3 expression were detected by immunohistochemistry. the association of IL-22 expression with FOXP3 tregs infiltration were evaluated. Results: Among 152 samples, 104(68.4%) cases exhibited IL-22 expression and 77(54.1%) cases exhibited high FOXP3 tregs infiltration. Both the increased IL-22 expression and elevated infiltration of FOXP3 tregs was positively associated with hormone receptor status, HER-2 status and molecular subtypes. Meanwhile, IL-22 expression and FOXP3 tregs infiltrations showed a positive correlation(r=0.250, P=0.0019). Among different molecular subtypes, HER2-enriched and basal-like types displayed both the higher IL-22 expression and FOXP3 tregs infiltration, and the tendency of FOXP3 tregs infiltration was in accordingly with IL-22 expression. Expression of IL-22 represented worse overall survival(OS), furthermore, IL-22 expression and FOXP3 tregs infiltration showed synergistic prognostic significance, patients with IL-22 positive expression together with high FOXP3 tregs infiltration displayed the much poorer clinical outcomes both in OS and relapse-free survival(RFS).Conclusions: Our data indicates that IL-22 expression and FOXP3 trges infiltration may work synergistically in breast cancer, and significantly correlated with patient prognosis, suggesting possible therapy targets and biomarkers in the future.

关键词： interleukin-22 breast cancer FOXP3 regulatory t cells HER-2 prognosis

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Analysis of regulatory t cells frequency in peripheral blood and tumor tissues in papillary thyroid carcinoma with and without Hashimoto’s thyroiditis

Analysis of regulatory T cells frequency in peripheral blood...

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2014第六届全国甲状腺肿瘤学术大会

作者： Xinwei Yun Youzhong Liu Ming Gao Yang Yu Xiaolong Li Department of Head ＆ Neck tumor Key Laboratory of Cancer Prevention and Therapy Tianjin Medical University Cancer Institute ＆ Hospital National Clinical Research Center of Cancer

Purpose: regulatory t cells（treg） suppress the immune reaction. the aim of the present study was to investigate the clinicopathologic significance and roles of treg in Papillary thyroid carcinoma（PtC） patients with and without Hashimoto’s thyroiditis. Methods: Flow cytometry was used to detect percentage of CD4+CD25+CD127low/-treg among CD4+ t cells in peripheral blood. Foxp3+ treg were detected by immunohistochemistry in the tumor tissues. Results: the percentage of CD4+CD25+CD127low/- treg among CD4+ t cells was significantly higher in PtC patients than that in thyroid nodular goiter（tNG） patients. there were large numbers of tumor-infiltrating Foxp3+ treg in primary PtC and metastasis lymph nodes tissues, however, there was no Foxp3 expression in the tNG tissues. Higher percentage of treg both in peripheral blood and tumor tissues was associated with extrathyroidal extension and lymph nodes metastasis. the percentage of CD4+CD25+CD127low/- treg among CD4+ t cells in peripheral blood of PtC patients with Hashimoto’s thyroiditis（Ht） was significantly lower, whereas the infiltration of Foxp3+ treg in tissues of PtC with Ht tended to be increased. Conclusions: We concluded that the prevalence of treg increased in peripheral blood as well as in the tumor tissues of PtC patients compared with that of tNG patients. the high prevalence of treg was associated with aggressiveness. there may be a compensatory expansion of treg at the sites of inflammation in tissues of PtC with Ht contributing the immune response suppression.

关键词： Papillary thyroid carcinoma regulatory t cells tumor escape Hashimoto’s thyroiditis tumor microenvironment

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Imbalanced signal transduction in FoxP3+ regulatory t cells

Imbalanced signal transduction in FoxP3+ regulatory T cells

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第十届全国免疫学学术大会

作者： Dapeng Yan Julia Farache Diane Mathis Christophe Benoist Department of Immunology School of Basic Medical Sciences Fudan University Division of Immunology Department of Microbiology and Immunobiology Harvard Medical School

regulatory t(treg) cells characterized by the transcription factor FoxP3 have a fundamental role in the establishment and maintenance of peripheral tolerance, and have transcriptional and functional phenotypes that distinguish them from conventional CD4+ t cells(tconv). there have been reports that these two CD4+ t cell subsets differ in the signaling downstream of t cell receptor-triggered activation, with different requirements for some signaling factors intermediates and lower responsiveness in treg cells. Seeking a more comprehensive view, we found that treg cells have a broadly dampened activation of several pathways and signaling nodes, with low phosphorylation of CD3ζ, SLP76, Erk1/2, AKt, or S6 and lower calcium influx, upon tCR-mediated activation. Examination of StAt responses to cytokines highlighted striking variations in treg vs tconv responses to interferons, IL2 or IL6, but no general treg signaling defect. treg/tconv differences in responses were not due to different tCR levels, but much of the difference could be attributed to lower responsiveness of cells with a CD44 or CD62 L cells, which form a greater proportion of the treg pool. testing putative candidate regulators, the treg/tconv differential could not be explained by treg over-expression of the signaling modulator CD5, the co-inhibitors PD- and CtLA4, or the regulatory phosphatase DUSP4. However, gene expression profiling in Dusp4-deficient mice showed that DUSP4 enhances the expression of a segment of the canonical treg transcriptional signature, a module which overlaps in part with the tCR-dependent treg gene set. taken together, our findings show treg cell signaling to be tuned down, plausibly because of their intrinsically higher reactivity to self, but comparatively more attuned to cytokines or other inter-cellular signals.

关键词： signal transduction regulatory t cells

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Complement component 3 deficiency prolongs MHC-Ⅱ disparate allograft survival by increasing the population of CD4+ CD25+ regulatory t cells

Complement component 3 deficiency prolongs MHC-Ⅱ disparate ...

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第十一届全国免疫学学术大会

作者： Zheng Quanyou Zhang Keqin Xu Guilian Department of Nephrology Southwest HospitalThird Military Medical University Department of Immunology Third Military Medical University

Objective: Recent reports suggest that both the complement system and alloreactive t cells contribute to allograft rejection. However, their underlying relationship and mechanism of action is poorly understood. Herein, we investigate the role of complement component 3(C3) in a single MHC- Ⅱ molecule mismatched murine model of allograft rejection using C3 deficient mice(C3) as skin graft donors or recipients.Methods: the allografts survival were monitored and histological changes of skin grafts were also assessed. Gene expression coding inflammatory cytokines and t cell proliferation in vitro were tested. the expansion of CD4CD25Foxp3 regulatory t cells and th1/17 cells were analyzed by Flow cytometry and immune histological staining. At last, the DC capacity was examined by co-cluture them with allo-reactive CD4t cells for up to 9 days.Results: Compared with C3/ B6 allografts, C3 B6 grafts dramatically prolonged survival in MHC-Ⅱ molecule mismatched H-2bm12 B6 recipients. However, no difference was observed when H-2bm12 B6 skin grafts were donated to C3/ B6 and C3 B6 recipients. Compared with C3/ allografts, both th17 cell infiltration and th1/th17 associated cytokine mR NA levels were clearly reduced in C3 allografts. Moreover, C3 allografts caused an impaired mixed lymphocyte reaction and attenuated th1/th17 responses, but increased CD4CD25Foxp3 regulatory t(treg) cell expression markedly in local intragraft and H-2bm12 recipients. Depletion of treg cells by anti-CD25 monoclonal antibody(m Ab) negated the survival advantages conferred by C3 deficiency. Additionally, C3 dendritic cells(DC) had more advantage in upregulating Foxp3 gene expression and treg cell induction compared to C3/ DC.Conclusion: Our results indicate for the first time that local C3 deficiency can prolong MHC-Ⅱ molecule mismatched skin allograft survival and confirm the association of local C3 deficiency with CD4 CD25 treg cell population expansion, which may then lead to impaired th1/th17 responses in recipients with C3 allografts.

关键词： Complement component 3 regulatory t cells Allograft rejection

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