分泌性中耳炎患者外周血Th1Th2细胞及T淋巴细胞亚群水平的表达及临床意义

目的:探讨分泌性中耳炎(SOM)患者外周血T辅助细胞1(Th1)、T辅助细胞2(Th2)细胞因子及T淋巴细胞亚群水平的表达及其临床意义。方法:选取我院于2015年1月至2018年1月期间收治的SOM患者135例记为SOM组,根据病程将患者分为急性组(病程14d,49例)、亚急性组(病程14-30d,53例)、慢性组(病程30d,33例)。另外选择同期于我院进行体检的100例健康者为对照组。分别对比SOM组和对照组受试者、不同病程SOM患者外周血Th1细胞因子[干扰素(INF-γ)、白细胞介素-2(IL-2)]、Th2细胞因子[白细胞介素-4(IL-4)、白细胞介素-10(IL-10)]以及T淋巴细胞亚群[CD3~+、CD4~+、CD8~+、CD4~+/CD8~+]水平。采用Pearson相关性分析INF-γ、IL-2、IL-4、IL-10与CD3~+、CD4~+、CD8~+、CD4~+/CD8~+的相关性。结果:SOM组患者外周血INF-γ、IL-2、IL-4、IL-10水平高于对照组(P0.05);急性组患者外周血INF-γ、IL-2、IL-4、IL-10水平低于亚急性组、慢性组,亚急性组患者外周血INF-γ、IL-2、IL-4、IL-10水平低于慢性组(P0.05)。SOM组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平低于对照组,CD8~+水平高于对照组(P0.05);急性组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平高于亚急性组、慢性组,CD8~+水平低于亚急性组、慢性组,亚急性组患者外周血CD3~+、CD4~+、CD4~+/CD8~+水平高于慢性组,CD8~+水平低于慢性组(P0.05)。Pearson相关性分析结果显示,SOM患者外周血INF-γ、IL-4与CD8~+呈正相关(P0.05),IL-4与CD3~+、CD4~+呈负相关(P0.05)。结论:SOM患者外周血Th1Th2细胞因子、T淋巴细胞亚群水平均表现异常,且其水平与疾病发生和发展存在一定联系,通过监测Th1Th2细胞因子、T淋巴细胞亚群有助于评估SOM患者病情。     

SLE带状疱疹患者外周血CD4~+CD28~+和CD4~+CD25~+FoxP3~+调节性T细胞的表达及相关性研究

目的:检测系统性红斑狼疮(systemic lupus erythematosus,SLE)合并带状疱疹患者外周血CD4~+CD28~+和CD4~+CD25~+Fox P3~+调节性T细胞的表达及相关性,探讨其在SLE合并带状疱疹发病中的临床意义。方法:采用流式细胞术检测30例SLE患者、30例SLE合并带状疱疹患者及30例健康对照者外周血中CD4~+/CD8~+T淋巴细胞亚群表面CD28的表达及CD4~+CD25~+Fox P3~+Treg细胞的表达水平,并分析SLE合并带状疱疹患者外周血CD4~+CD28~+和CD4~+CD25~+Fox P3~+调节性T细胞表达的相关性。结果:SLE合并带状疱疹组患者急性期外周血CD4~+T淋巴细胞比率、绝对计数显著降低,CD4~+、CD8~+T淋巴细胞表面的CD28表达下调,CD4~+CD25~+Fox P3~+Treg细胞水平显著高于SLE组及健康对照组,SLE合并带状疱疹组患者外周血CD4~+CD25~+Fox P3~+Treg水平与CD4~+CD28~+水平成负相关(P均0.05)。结论:SLE合并带状疱疹患者CD4~+、CD8~+T细胞活化异常,CD4~+CD25~+Fox P3~+Treg细胞可能参与抑制了T细胞的活化。     

小细胞肺癌患者外周血淋巴细胞亚群分析

目的 探究化疗对小细胞肺癌(small cell lung cancer,SCLC)患者免疫功能的影响。 方法 选择2013年1月到2018年12月我院收治的95例小细胞肺癌患者为研究对象。患者第一周期、第二周期化疗前采用流式细胞术检测患者外周血淋巴细胞亚群水平,分别按照不同疗效及不同化疗方案对患者外周血淋巴细胞亚群进行比较。 结果 (1)化疗后,95例患者CD3+、CD4+、CD8+细胞平均值增加,CD19+、γδT细胞平均值减少,差异均有统计学意义(均P+、CD8+细胞平均值增加,CD19+细胞减少,差异有统计学意义(均P0.05)。(3)依托泊苷联合顺铂(EP)方案组化疗后患者CD3+、CD8+细胞平均值增多,CD19+细胞减少,差异均有统计学意义(均P0.05)。 结论 化疗可以调节小细胞肺癌患者的免疫功能,增强细胞免疫,降低体液免疫,其中EC方案对患者细胞免疫的增强作用较为显著。     

肺癌患者外周血淋巴细胞亚群水平的表达及临床意义

目的:探讨肺癌患者外周血淋巴细胞亚群水平的表达及临床意义。方法:选择2016年3月~2017年3月期间我院收治的88例肺癌患者作为研究组,选择同期于我院进行健康体检的88例受检者作为对照组。两组研究对象均通过流式细胞仪检测外周血T淋巴细胞亚群水平。观察对比两组研究对象外周血T淋巴细胞亚群的表达水平,以及研究组不同临床病理特征患者外周血T淋巴细胞亚群的表达水平。结果:研究组CD4~+/CD8~+、CD4~+、CD3~+表达水平均低于对照组,CD8~+表达水平高于对照组,差异具有统计学意义(P0.05)。研究组Ⅲ期与Ⅳ期患者CD4~+/CD8~+、CD4~+、CD3~+表达水平均低于Ⅰ期与Ⅱ期,Ⅲ期与Ⅳ期患者CD8~+表达水平高于Ⅰ期与Ⅱ期(P0.05);小细胞肺癌患者CD4~+/CD8~+、CD4~+、CD3~+表达水平均低于非小细胞肺癌患者,CD8~+表达水平高于非小细胞肺癌患者,差异具有统计学意义(P0.05)。不同性别、年龄、肿瘤分化程度肺癌患者CD4~+/CD8~+、CD8~+、CD4~+、CD3~+水平无统计学差异(P0.05)。结论:肺癌患者外周血淋巴细胞亚群水平的表达呈现异常状态,且表达水平与疾病的分期和病理分型有关。     

CD4+ T细胞亚群在支气管哮喘病理机制中作用的新认识

CD4+T细胞对病原微生物可产生有效免疫应答,依据其产生细胞因子不同分为辅助Ⅰ型T细胞(Th1)和辅助Ⅱ型T细胞(Th2),目前其他亚型已被鉴定,包括Th17、调节性T细胞(Treg),滤泡型T细胞和Th9细胞等.调节性T细胞既抑制Th1,也抑制Th2发育.最近研究报道,在过敏性疾病和哮喘患者中Treg细胞数量减少,并认为是通过下调Th1和Th2细胞以外的其他亚类细胞而抑制过敏和哮喘发作的.在这篇综述中,我们将进一步探讨Th1、Th2、Th17、Th9以及Treg在过敏性和哮喘疾病中的作用.     

可溶性Jagged-1/Fc嵌合蛋白诱导淋巴结细胞向CD4+CD25+ T细胞分化

直接用可溶性Jagged-1/Fc嵌合蛋白(Jagged-1/Fc)在体外诱导小鼠淋巴结细胞向CD4 CD25 T细胞分化.通过荧光标记单克隆抗体染色结合流式细胞术,观察不同剂量Jagged-1/Fc在不同时间对淋巴结细胞向CD4 CD25 T细胞分化的影响,观察Jagged-1/Fc诱导T细胞内细胞因子的变化;藉ELISA法检测Jagged-1/Fc诱导分化的T细胞分泌TGF-β1、IL-4和IL-10的水平.结果显示,超过500.0μg/L剂量的Jagged-1/Fc使CD4 CD25 T细胞百分比明显增高,诱导时间需要4～6天,抗Jagged-1单抗能抵消Jagged-1/Fc的诱导作用,用DAPT阻断Notch信号通路的活化也能抑制Jagged-1/Fc的诱导作用,Jagged-1/Fc诱导分化的T细胞培养上清中IL-4和IL-10的水平明显增高,TGF-β1无明显变化,胞内IL-4,IL-10,IL-2和TNF-α的水平也呈增高趋势.上述结果表明,可溶性Jagged-1/Fc嵌合蛋白在体外可诱导小鼠淋巴结细胞向CD4 CD25 调节性T细胞分化.     

非霍奇金淋巴瘤患者外周血中CD4+CD25+ 调节性T 细胞 亚群变化初探

目的:检测非霍奇金淋巴瘤(non-Hodgkin’s lymphoma,NHL)患者外周血中CD4+CD25+调节性T细胞(CD4+CD25+regulatoryT cell,Treg)的改变,探讨Treg与NHL的相关性。方法:病例组(n=60)为本院收治的初诊NHL患者,对照组(n=60)为本院健康体检者,用流式细胞技术联合标记CD4、CD25检测对照组及病例组化疗前、化疗后的外周血中CD4+CD25+调节性T细胞的分布特点。结果:(1)病例组化疗前外周血中CD4+细胞比例显著低于对照组(P<0.05),CD4+CD25+调节性T细胞比例显著高于对照组(P<0.05);(2)病例组化疗后,CD4+细胞比例明显高于化疗前(P<0.05),CD4+CD25+调节性T细胞比例明显低于化疗前(P<0.05);(3)病例组化疗后CD4+细胞比例与对照组无显著差异(P>0.05),而CD4+CD25+调节性T细胞比例显著高于对照组(P<0.05)。结论:非霍奇金淋巴瘤患者外周血中CD4+CD25+调节性T细胞比例升高,存在机体免疫抑制,化疗可降低CD4+CD25+调节性T细胞比例。     

中药复方连黄对小鼠T淋巴细胞亚群CD4~+、CD8~+的影响

目的探讨中药复方连黄对小鼠T淋巴细胞亚群CD4+、CD8+的影响.方法 T淋巴细胞亚群测定采用单克降抗体直接免疫荧光技术,通过流式细胞仪测定.结果经统计学分析,与对照组比较,中药复方连黄能不同程度地使CD4+、CD4+/CD8+升高,而使CD8+下降.结果表明,中药复方连黄对细胞免疫功能具有调节作用.     

银屑病和SLE患者T细胞亚群与淋巴细胞CD2分子的相关性研究

目的：研究银屑病和系统性红斑狼疮（systemic lulups erythematosus,SLE）患者T淋巴细胞CD2、CD4＋、CD8＋以及CD4＋/CD8＋的变化及其相关性。方法：应用流式细胞仪检测我院收治的62例银屑病和31例SLE患者T淋巴细胞CD2、CD4＋、CD8＋及CD4＋/CD8＋分子的表达水平并统计学分析其相关性。结果：银屑病患者T淋巴细胞CD4＋/CD8＋的比值与正常对照组比较显著增高,而SLE患者与正常对照组比较显著降低。银屑病患者CD4＋百分率与正常对照组比较显著增高,SLE患者CD8＋与正常对照组比较显著增高。银屑病患者CD2与CD4＋之间呈正相关（γ=0.309,P〈0.05）,CD4＋与CD4＋/CD8＋之间呈显著正相关（γ=0.536,P〈0.05）,SLE患者CD2仅与CD4＋之间呈正相关（γ=0.378,P〈0.05）。结论：T淋巴细胞免疫功能紊乱可能在银屑病的发病机制中发挥了重要作用,CD2可能使银屑病的T淋巴细胞紊乱致炎症加重。SLE患者T细胞亚群CD4＋/CD8＋比值下降,CD2与CD4呈正相关亦可能使SLE的T淋巴细胞加速信号传导紊乱,CD8＋百分率增高可能在SLE的发病机制中起重要作用。     

慢性阻塞性肺疾病患者外周血CD4+CD25+CD127(Low/-)调节性T细胞的检测及临床意义

目的:研究CD4+CD25+CD127(Low/-)节性T细胞在慢性阻塞性肺疾病(COPD)急性发作期外周血中的比例改变及其临床意义.方法:以25例COPD急性发作期患者外周血为研究组,20名正常人外周血作为对照组,采用三色直接荧光素标记法和多参数流式细胞仪检测外周血CD4+CD25+CD127(Low/-)调节性T细胞的比例,同时检测外周血C-反应蛋白(CRP)、血沉(ESR)、免疫球蛋白(Ig)等水平.结果:COPD急性发作期患者外周血CD4+CD25+CD127(Low/-)调节性T细胞占外周血CD4+淋巴细胞的比例明显低于健康对照组(P＜0.01).而COPD急性发作期患者外周血CRP、ESR、Ig等水平明显高于健康对照组(P＜0.05).COPD患者外周血调节T细胞下降与CRP和IgG升高成负相关.结论:COPD患者外周血CD4+CD25+CD127(Low/-)调节性T细胞在CD4+T淋巴细胞的比例明显减少,调节性T细胞等免疫调节因素可能在COPD的发病机制发挥重要作用.     

In vitro expansion of CD4+CD25highFOXP3+CD127low/- regulatory T cells from peripheral blood lymphocytes of healthy Mycobacterium tuberculosis-infected humans

CD4+CD25highFOXP3+ regulatory T (Treg) cells have recently been found at elevated levels in the peripheral blood of tuberculosis patients, compared to Mycobacterium tuberculosis latently infected (LTBI) healthy individuals and non-infected controls. Here, we show that CD4+CD25highFOXP3+ T lymphocytes can be expanded in vitro from peripheral blood mononuclear cells (PBMC) of LTBI individuals, but not of uninfected controls by incubating them with BCG in the presence of TGF-beta. These expanded cells from the PBMC of LTBI subjects expressed CTLA-4, GITR and OX-40, but were CD127low/- and have therefore the phenotype of Treg cells. In addition, they inhibited in a dose-dependant manner the proliferation of freshly isolated mononuclear cells in response to polyclonal stimulation, indicating that they are functional Treg lymphocytes. In contrast, incubation of the PBMC with BCG alone preferentially induced activated CD4+ T cells, expressing CD25 and/or CD69 and secreting IFN-gamma. These results show that CD4+CD25highFOXP3+ Treg cells can be expanded or induced in the peripheral blood of LTBI individuals in conditions known to predispose to progression towards active tuberculosis and may therefore play an important role in the pathogenesis of the disease.     

Human CD4+CD25+ regulatory T cells inhibit the differentiation of osteoclasts from peripheral blood mononuclear cells

Regulatory T cell (Treg) is a subset of CD4+ T lymphocytes expressing CD25 with immunosuppressive activity. However the function of Tregs onto osteoclastogenesis remains unknown. We investigated the effect and regulatory mechanism of Treg focusing on osteoclastogenesis from PBMCs. Tregs were isolated from PBMCs by magnetic cell sorting-column and analyzed by flow cytometry. RT-PCR was performed to identify Foxp3 mRNA. Using PBMCs and Tregs coculture system, we could find that Tregs inhibited osteoclasts differentiation from PBMCs and reduced the resorbed areas on pit assay (p <0.01). This suppression of osteoclast differentiation was cytokine-dependent, not cell-to-cell direct contact proved by Transwell system. Tregs-induced osteoclast differentiation was blocked by anti-TGF-beta or anti-IL-4 antibody treatment. These results suggest that Tregs inhibit osteoclast differentiation from PBMCs in a cytokine-dependent manner, not by cell-to-cell contact manner and that TGF-beta and IL-4 may be the key cytokines for this suppressive function of Tregs.     

CD4+CD25+ 调节性T 细胞与冠状动脉粥样硬化病变的研究进展

CD4+CD25+调节性T细胞是一个具有独特免疫调节功能的T细胞亚群,人体主要通过CD4+CD25+调节性T细胞以免疫负向调节的方式来抑制自身反应性T细胞的作用,减少免疫性疾病的发生,从而维持机体内环境的稳定,维持免疫耐受。CD4+CD25+Treg已被证实其与肿瘤、感染、自身免疫病、移植免疫等多种疾病的发生、发展及转归均相关。随着社会的进步和人民生活水平的提高冠状动脉粥样硬化性病变作为一种慢性病变,其发病率越来越高,已经成为严重危害人类健康的常见病,近年来越来越多的证据表明炎症及免疫反应机制在冠状动脉粥样硬化性心脏病的发生、发展及预后过程中具有重要的作用。而CD4+CD25+调节性T细胞在冠状动脉粥样硬化性病变中所起的作用也受到越来越多的关注。本文就CD4+CD25+调节性T细胞与冠状动脉粥样硬化病变之间的关联做一综述。     

益生菌联合布地奈德对支气管哮喘患儿的疗效及对外周血CD4+和CD8+细胞水平的影响

研究益生菌联合布地奈德对支气管哮喘患儿的疗效及对外周血CD4⁺和CD8⁺细胞水平的影响, 为该类患者的治疗提供参考。

选择2019年6月至2020年5月我院收治的92例支气管哮喘患儿为研究对象, 按照随机数字表法分为观察组(益生菌联合布地奈德治疗)和对照组(布地奈德治疗)各46例, 比较两组患儿疗效、临床症状消失时间、治疗前及治疗4周后T淋巴细胞(CD4⁺、CD8⁺)和肺功能[用力肺活量(FVC)、第1秒用力呼气容积(FEV1)、最大呼气峰流速(PEF)]水平。

治疗4周后, 两组患儿总有效率比较差异无统计学意义(93.48%vs 89.13%, χ²=0.137, P=0.711)。观察组患儿咳嗽消失时间、气促消失时间、哮鸣音消失时间、憋喘消失时间均显著低于对照组(均P < 0.05)。治疗4周后, 观察组患儿CD4⁺细胞水平显著低于治疗前及同期对照组, CD8⁺细胞、肺功能(FVC、FEV1、PEF)水平显著高于治疗前及同期对照组(均P < 0.05)。

益生菌联合布地奈德对支气管哮喘的疗效显著, 能够促进患儿临床症状恢复, 改善患儿肺功能, 下调外周血CD4⁺细胞水平, 上调CD8⁺细胞水平。

     

Characterization of a murine thymic CD4+ T cell subset-TCRαβ+ 3G11-6C10- CD4+ CD8-thymocytes

The presence of a relatively mature CD4⁺ CD8⁻ (SP) T cell subset in mouse thymus has been demonstrated. Composing of 10% of total CD4SP thymocytes, this subset is defined by the absence of 3G11 and 6C10 expression with a phenotype of CD69^+/−, HSA^med/lo and heterogeneous for Qa-2 expression. The proliferation capability of TCRαβ⁺ 3Gl l⁻ 6C10⁻ CD4⁺ CD8⁻ thymocytes was high while using Con A stimulus. And Con A stimulation could result in secretion of IL4, IL-10, IL-6 and a little amount of IFNγ. IL-2 was barely detectable. This is distinct from typical Th0 type cytokines. The cells of this subset were NK1.1 negative, but strongly expressed GATA-3 mRNA. The results suggest that the CD4⁺ subset of 3G11⁻ 6C10⁻ NK1.1⁻ phenotype possesses immunocompetent cells with functions characteristic of Th2-like cytokines, which may indicate the cells at transitional status from Th0 to Th2, with a propensity to Th2. Project supported by the National Natural Science Foundation of China (Grant No. 39730410).     

Peripheral blood CD4+CD25+CD127low regulatory T cells are significantly increased by tocilizumab treatment in patients with rheumatoid arthritis: increase in regulatory T cells correlates with clinical response

IntroductionTocilizumab (TCZ), an anti-interleukin-6 receptor antibody, is clinically effective against rheumatoid arthritis (RA), and several reports have indicated how TCZ influences a number of mechanisms underlying RA pathogenesis. However, it is still unclear whether TCZ affects inflammatory cells in peripheral blood and whether any such changes are associated with clinical response. We evaluated associations between proportions of subsets of peripheral immune cells and clinical response in patients with RA treated with TCZ.MethodsThirty-nine consecutive patients with RA who started to receive TCZ as their first biologic between March 2010 and April 2012 were enrolled. The proportions of several subsets of peripheral cells with their levels of expression of differentiation markers, activation markers and costimulatory molecules were measured sequentially from baseline to week 52 by flow cytometry analysis.ResultsClinical Disease Activity Index (CDAI) remission was achieved in 53.8% of patients at week 52 of TCZ therapy. The proportions of CD4⁺CD25⁺CD127^low regulatory T cells (T_reg) and HLA-DR⁺ activated T_reg cells significantly increased with TCZ therapy (P < 0.001 and P < 0.001, respectively), whereas proportions of CD3⁺CD4⁺CXCR3⁻CCR6⁺CD161⁺ T helper 17 cells did not change over the 52 weeks. The proportions of CD20⁺CD27⁺ memory B cells, HLA-DR⁺CD14⁺ and CD69⁺CD14⁺ activated monocytes, and CD16⁺CD14⁺ monocytes significantly decreased (P < 0.001, P < 0.001, P < 0.001 and P < 0.001, respectively). Among them, only the change in T_reg cells was inversely correlated with the change in CDAI score (ρ = −0.40, P = 0.011). The most dynamic increase in T_reg cells was observed in the CDAI remission group (P < 0.001).ConclusionThis study demonstrates that TCZ affected proportions of circulating immune cells in patients with RA. The proportion of T_reg cells among CD4⁺ cells correlated well with clinical response.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-015-0526-4) contains supplementary material, which is available to authorized users.     

Expansion of CD4+CD25+ helper T cells without regulatory function in smoking and COPD

Background

Regulatory T cells have been implicated in the pathogenesis of COPD by the increased expression of CD25 on helper T cells along with enhanced intracellular expression of FoxP3 and low/absent CD127 expression on the cell surface.

Method

Regulatory T cells were investigated in BALF from nine COPD subjects and compared to fourteen smokers with normal lung function and nine never-smokers.

Results

In smokers with normal lung function, the expression of CD25⁺CD4⁺ was increased, whereas the proportions of FoxP3⁺ and CD127⁺ were unchanged compared to never-smokers. Among CD4⁺ cells expressing high levels of CD25, the proportion of FoxP3⁺ cells was decreased and the percentage of CD127⁺ was increased in smokers with normal lung function. CD4⁺CD25⁺ cells with low/absent CD127 expression were increased in smokers with normal lung function, but not in COPD, when compared to never smokers.

Conclusion

The reduction of FoxP3 expression in BALF from smokers with normal lung function indicates that the increase in CD25 expression is not associated with the expansion of regulatory T cells. Instead, the high CD127 and low FoxP3 expressions implicate a predominantly non-regulatory CD25⁺ helper T-cell population in smokers and stable COPD. Therefore, we suggest a smoking-induced expansion of predominantly activated airway helper T cells that seem to persist after COPD development.