宫颈癌患者外周血CD4~+CD25~+调节性T淋巴细胞与白细胞介素-17水平及其相关性研究

目的探讨外周血CD4~+CD25~+调节性T淋巴细胞(Tr细胞)和白细胞介素-17(IL-17)水平在宫颈癌患者中的变化及其相关性。方法选取2014年1月至2015年3月该院确诊宫颈癌患者60例(宫颈癌组)和查体的健康女性60例(健康对照组),采用流式细胞仪检测外周血Tr细胞数量及占CD4~+细胞百分比,采用酶联免疫吸附试验(ELISA)检测血清IL-17水平。结果宫颈癌组外周血Foxp3~+Tr、Tr、CD4~+T淋巴细胞百分比,Foxp3~+Tr/Tr、Tr/CD4~+T淋巴细胞比值及血清IL-17水平均较健康对照组明显升高,而IL-17/Tr比值明显下降,差异有统计学意义(P0.05)。宫颈癌组患者外周血CD4~+CD25~+Tr细胞水平与血清IL-17水平呈正相关(r=0.768,P0.05)。结论宫颈癌患者存在细胞免疫功能紊乱,IL-17与Tr细胞比值失衡可能在宫颈癌发病机制中起一定的作用。     

慢性乙型肝炎患者外周血CD8+CD25+FoxP3+调节性T细胞比例的变化及临床意义

目的 探讨外周血CD8+CD25+FoxP3+调节性T细胞(Treg)比例在慢性乙型肝炎(CHB)患者中的变化及临床意义.方法 选取2018年3-11月在滨海县第二人民医院传染科诊治的无症状乙型肝炎病毒(HBV)携带者28例为携带组,CHB患者28例为CHB组.同时选取28例年龄、性别匹配的健康体检者作为对照组.采用流式细胞仪分析外周血CD8+CD25+FoxP3+Treg比例.比较各组肝功能相关指标、HBV-DNA、外周血CD8+CD25+FoxP3+Treg比例、外周血单个核细胞(PBMC)中叉头状转录因子P3(FoxP3)mR-NA水平,以及血清白细胞介素(IL)-10、IL-35、转化生长因子(TGF)-β1水平.分析CHB患者外周血CD8+CD25+FoxP3+Treg比例与肝功能相关指标、HBV-DNA、PBMC中FoxP3 mRNA水平,以及血清IL-10、TGF-β1、IL-35水平的相关性.结果 与对照组、携带组相比,CHB组总胆红素(TBIL)、直接胆红素、丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶水平均明显升高,差异有统计学意义(P<0.05).与对照组相比,携带组、CHB组外周血CD8+CD25+FoxP3+Treg比例、PBMC中FoxP3 mRNA水平及血清IL-10、TGF-β1、IL-35水平均明显升高,且C HB组均高于携带组,差异有统计学意义(P<0.05).C HB患者外周血CD8+CD25+FoxP3+Treg比例与PBMC中FoxP3 mRNA水平,血清IL-10、TGF-β1及IL-35水平均呈正相关(r=0.568、0.537、0.377、0.484,P<0.05);CHB患者外周血CD8+CD25+FoxP3+Treg比例与TBIL、HBV-DNA、ALT水平呈正相关(r=0.536、0.570、0.443,P<0.05).结论 外周血CD8+CD25+FoxP3+Treg比例升高可能在C HB患者疾病发展过程中发挥了重要作用.     

食管癌患者外周血CD3+T细胞分泌细胞因子的水平变化及其临床意义

目的探讨食管癌患者外周血CD3~+T细胞分泌细胞因子的水平变化及其临床意义。方法选取2016年1月至2018年1月在西安交通大学第二附属医院接受治疗的28例食管癌患者为研究组,28例健康体检者为健康对照组。比较两组患者的外周血T细胞、外周血CD3~+T细胞分泌因子白细胞介素(IL)-2、IL-4、IL-10、IL-12表达水平、外周血CD3~+T细胞分泌因子肿瘤坏死因子(TNF)-α和干扰素(IFN)-γ表达水平。结果研究组CD3~+/CD4~+、CD4~+/CD8~+表达水平低于对照组,研究组CD3~+/CD8~+、CD3~+表达水平高于对照组,差异有统计学意义(P0.05)。研究组IL-2、IL-10、IL-12表达水平低于对照组,研究组IL-4表达水平高于对照组,差异有统计学意义(P0.05)。研究组TNF-α表达水平高于对照组,研究组IFN-γ表达水平低于对照组,差异有统计学意义(P0.05)。结论对于食管癌患者,食管癌患者外周血CD3~+T细胞分泌细胞因子CD4~+/CD8~+、CD3~+/CD4~+表达水平显著增高,说明患者的免疫耐受力与免疫逃逸存在较明显的关系。     

IL-6、IL-8、CD64和CD11b检测对新生儿感染性疾病的诊断价值

目的探讨白细胞介素-6(IL-6)、IL-8、CD64和CD11b在新生儿感染性疾病早期诊断中的价值。方法选取2013年3月至2014年3月上海市第一人民医院儿科和产科收治的新生儿,分为感染组(50例)、非感染对照组(30例)、健康对照组(50例);采用定量夹心酶联免疫吸附试验(ELISA)测定血清IL-6和IL-8水平,采用全血三色荧光标记法流式细胞术检测外周血中性粒细胞表面CD64和CD11b分子表达水平,比较分析3组新生儿的IL-6、IL-8、CD64和CD11b水平及阳性率。结果对于血清IL-6、IL-8两项检测指标,感染组分别为(187.3±25.1)ng/L、(1.05±0.32)pg/mL,非感染对照组分别为(60.9±5.2)ng/L、(0.35±0.12)pg/mL,健康对照组分别为(53.2±9.3)ng/L、(0.30±0.05)pg/mL;感染组分别高于非感染对照组和健康对照组,差异均有统计学意义(P0.05)。并且感染组中性粒细胞表面CD64、CD11b分子表达水平分别高于非感染对照组和健康对照组,差异均有统计学意义(P0.05)。结论临床检测新生儿血清IL-6、IL-8水平及外周血中性粒细胞表面CD64和CD11b分子的表达水平,可更加及时、准确、灵敏地诊断新生儿早期感染性疾病。     

胎盘CD68、PD-1、IL-10与妊娠并HCV感染关系及对母婴结局的影响

目的探究胎盘CD68、程序性死亡受体-1(PD-1)、白细胞介素10(IL-10)与妊娠并丙型肝炎病毒(HCV)感染关系及对母婴结局的影响。方法选取本院2016年10月至2019年10月妊娠并HCV感染患者116例作为观察组,同期非HCV感染妊娠孕妇58例作为对照组。比较两组、不同HCV病毒载量患者胎盘CD68、PD-1阳性细胞数及IL-10水平,分析三者与HCV病毒载量的关系;并对比两组间、观察组胎盘不同CD68、PD-1、IL-10表达患者母婴结局。结果观察组胎盘CD68、PD-1阳性细胞数低于对照组,IL-10高于对照组,差异有统计学意义(U=7.258、6.346、15.669,P<0.05);胎盘CD68、PD-1高表达患者产妇新生儿不良结局发生率低于低表达患者,IL-10高表达患者产妇新生儿不良结局发生率高于低表达患者,差异有统计学意义(c2=7.107、9.175、7.306,P<0.05);胎盘CD68、PD-1阳性细胞数与HCV病毒载量呈负相关,IL-10水平与HCV病毒载量呈正相关(r=-0.741、-0.644、0.561,P<0.05)。结论胎盘CD68、PD-1表达下降、IL-10表达升高可能参与胎盘抗病毒免疫,从而对母婴结局产生不良影响。     

大鼠肾移植急性排斥反应调节性T细胞及其基因表达的机制研究

目的观察肾移植大鼠外周血中CD4+CD25+调节性T细胞及其表面标志物Foxp3、ABCC4、e IF6基因表达,结合白细胞介素-2(IL-2)和肿瘤坏死因子-γ(INF-γ)水平及肝、肾功能变化进行相关分析,探讨其在移植免疫排斥机制中的作用及意义。方法监测急排组与非急排组大鼠肝、肾功能;ELISA方法检测血浆中IL-2和INF-γ的表达水平;流式细胞仪检测CD4+、CD8+、CD4+CD25+Treg细胞亚群;荧光PCR检测CD4+CD25+Treg细胞Foxp3、ABCC4及e IF6 m RNA表达。结果与术前相比,急排组术后第7天血清肌酐、尿素氮均明显升高(P<0.05);急排组术后第7天IL-2及IFN-γ均升高,差异有统计学意义(P<0.05);急排组术后CD3+、CD4+、CD8+水平明显升高,差异有统计学意义(P<0.05);与非急排组相比,急排组术后CD4+CD25+Treg细胞表达降低,差异有统计学意义(P<0.05)。与非急排组相比,急排组术后Foxp3+基因表达量明显降低,ABCC4、e IF6基因表达量升高,差异有统计学意义(P<0.05)。结论 CD4+CD25+调节性T细胞在肾移植排斥反应中起到重要作用,CD4+CD25+Treg细胞Foxp3、ABCC4及e IF6 m RNA参与急性排斥反应的发生,检测其变化可辅助诊断急性排斥反应的发生。     

类风湿性关节炎患者CD8~+ CD122~+调节性T淋巴细胞和白细胞介素-10的水平变化及意义

目的探讨类风湿性关节炎(RA)患者外周血中CD8+CD122+调节性T淋巴细胞(Tregs)及细胞因子白细胞介素-10(IL-10)的水平变化及其临床意义。方法收集62例RA患者与26例健康体检者,采用流式细胞仪测定研究对象外周血中的CD8+CD122+Tregs水平,双抗夹心酶联免疫吸附试验测定血清中IL-10浓度。结果RA活动期和稳定期患者外周血中的CD8+CD122+Tregs和IL-10的含量明显低于健康对照组,差异有统计学意义(P<0.05)。RA活动期患者上述指标含量均显著低于稳定期患者,差异均有统计学意义(P<0.05)。同时,RA患者血清中的IL-10的浓度与外周血中的CD8+CD122+Tregs细胞水平情况呈正相关(rs=0.297,P<0.05)。结论 RA患者外周血中具有免疫负调控功能的CD8+CD122+Tregs和IL-10水平较低,可能与类风湿性关节炎的发生相关。     

淋巴结核患者外周血CD4~+CD25~(high)FoxP3~+调节性T淋巴细胞以及血浆IFN-γ和IL-10水平及其临床意义

目的研究淋巴结核患者外周血CD4+CD25highFoxP3+调节性T淋巴细胞(Treg)和干扰素γ(IFN-γ)、白细胞介素10(IL-10)水平的变化及临床意义。方法采用流式细胞仪对100例淋巴结核患者及30名正常人外周血CD4+CD25highFoxP3+Treg和IFN-γ、IL-10水平进行检测。结果淋巴结核患者组与正常对照组比较,外周血CD4+CD25highFoxP3+Treg和IL-10水平均升高(P0.05),而IFN-γ水平则降低(P0.05)。在干酪样型、增殖型和混合型3种类型的淋巴结核病之间各指标差异均无统计学意义(P0.05)。结论淋巴结核患者细胞免疫功能明显异常。CD4+CD25highFoxP3+Treg和IFN-γ、IL-10在淋巴结核的发病过程中发挥了重要作用。     

再生障碍性贫血患者外周血CD3 CD4 CD8的表达及造血相关因子水平的研究

目的研究再生障碍性贫血患者外周血中T细胞亚群及白细胞介素-2(IL-2)、干扰素-r(IFN-r)、肿瘤坏死因子(TNF-a)、白细胞介素-8(IL-8)、IL-12等造血细胞因子水平,旨在探讨再障患者的免疫状态。方法25例再障患者和15例正常对照者造血相关因子采用ELISA法。CD3、CD4、CD5荧光标记均用直接免疫法以流式细胞仪检测。结果SAA组患者CD3、CD4、C08细胞平均值分别为(69.41±6.30)%(、31.88±7.07)%、(35.17±6.26)%,与对照组比较,有显著性差异(分别为P<0.0l,P<0.01)。SAP,组IL-2、IFN-T、IFN-a、IL-8、IL-12中,除TNF-a外,其余均显著高于对照组(P<0.01),其中IFN-T、IL-8显著高于CM组(P<0.05);CAA组IL-2、IFN-T、IL-12高于对照组(分别为P<0.05,P<0.05,P(0.01),TNF-a、IL-8与对照组比较差异无显著性(P>0.05)。CAI组及SM组IL-12(P70)均显著高于正常对照组,差异有显著性(均为P<0.01),SAP,组IL-12(P70)水平高于CIA组,但无显著性差异(P>0.05)。IL-12(P70)与CD8 细胞%、IFN-T、IL-2呈正相关,与CD,细胞%呈负相关。结论大部分再障患者存在细胞免疫异常,以T淋巴细胞亚群失调及多种造血负调控因子异常增高为特征,与AA的发病有密切关系。     

移植患者CD4+CD25+CD127low调节性T细胞、TGF-β1及IL-10的表达

目的观察肝肾移植患者外周血中调节性T细胞(Treg细胞)的百分比与转化生长因子β1(TGF-β1)及白细胞介素10(IL-10)水平的表达情况及其相关性。方法15例移植患者的外周静脉血,采用流式细胞术检测CD4^+CD25^+CD127^low Treg细胞的百分比,并以20例体检健康者为对照组;用酶联免疫吸附试验法检测移植患者血清中细胞因子TGF-β1、IL-10的水平,分析这2种细胞因子水平与CD4^+CD25^+CD127^low Treg细胞百分比的关系及移植前后TGF-β1、IL-10水平的差异。结果与对照组比较,移植患者移植前CD4^+CD25^+CD127^low Treg细胞占CD4+T淋巴细胞的百分比明显升高,差异有统计学意义(P<0.05);肝肾移植后CD4^+CD25^+CD127^low Treg细胞百分比下降为0。肝肾移植前CD4^+CD25^+CD127^low Treg细胞百分比与TGF-β1、IL-10的水平呈正相关(P<0.05)。TGF-β1、IL-10的水平移植后低于移植前,差异无统计学意义(P>0.05)。结论CD4^+CD25^+CD127^low Treg的百分比与TGF-β1、IL-10的水平存在一定相关性;移植后CD4^+CD25^+CD127^low Tre细胞百分比下降为0,可能与应用大量免疫抑制剂有关。     

Tyrosine phosphorylation of CD6 by stimulation of CD3: augmentation by the CD4 and CD2 coreceptors

When T cells are activated via the T cell receptor (TCR) complex a number of cellular substrates, including some cell surface proteins, become phosphorylated on tyrosine (Tyr) residues. Phosphorylation of cytoplasmic Tyr renders these cell surface receptors competent to interact with proteins that link cell surface receptors to protein in the intracellular signaling pathways. Here we show that Tyr residues in the cytoplasmic domain of CD6 become phosphorylated upon T cell activation via the TCR complex. Tyr phosphorylation was observed when the T cells were activated by crosslinking CD3 or by cocrosslinking CD3 with CD2 or CD4, but not when the cells were stimulated by crosslinking CD2, CD4, or CD28 alone. Unlike other Tyr kinase substrates, such as the phospholipase C gamma 1-associated pp35/36 protein, whose level of Tyr phosphorylation is highest when T cells are activated by cocrosslinking CD3 with CD2, the levels of CD6 Tyr phosphorylation are highest when T cells were activated by cocrosslinking CD3 with CD4.     

CD137 enhances cytotoxicity of CD3CD56 cells and their capacities to induce CD4 Th1 responses

CD137 (4-1BB) is a TNFR superfamily member that mediates the costimulatory signal resulting in T cells and NK cells proliferation and cytokines production, but the effects of CD137 signaling on CD3⁺CD56⁺ cell subpopulation have not been well-documented. The aim of this study was to investigate the effects of CD137 signaling on regulation of CD3⁺CD56⁺ cell function. Anti-CD137 mAb or mouse IgG1 isotype control was added to CIK cell culture to determine the effects of proliferation and anti-tumor effects on CD3⁺CD56⁺ cells. We observed that anti-CD137 mAb could dramatically promote proliferation of CIK cells. And CD137–CIK cells and CD3⁺CD56⁺ cell subpopulation within them possessed higher ability to kill tumor cell line A549. The SCID mice engrafted with A549 cells and treated with CD137–CIK cells have prolonged survival. Further studies revealed that the percentages of CD3⁺CD56⁺ cells were elevated significantly in CD137–CIK cells. The expression of NKG2D was up-regulated on CD3⁺CD56⁺ cells from CD137–CIK cells. The expression of IFN-γ, IL-2 and TNF-α increased significantly whereas the production of TGF-β₁, IL-4 and IL-10 decreased in CD3⁺CD56⁺ cells from CD137–CIK cells. In addition, anti-CD137 mAb can elevate the capacity of CD3⁺CD56⁺ cells to induce CD4⁺ Th1 responses. We further showed that the anti-CD137 mAb also had the same effects on CD3⁺CD56⁺ cells expanded from the PBMCs of patients with NSCLC. We concluded that CD137 signaling could enhance the abilities of CIK cells to kill tumor cells in vitro and in vivo via increasing the proportion of CD3⁺CD56⁺ cells and their cytotoxicity. Furthermore, CD137 signaling can elevate the capacity of CD3⁺CD56⁺ cells to induce CD4⁺ Th1 responses which may enhance their anti-tumor activity indirectly. Taken together, our studies could be considered as valuable in CIK cells-based cancer immunotherapy.     

CD4+CD25+和CD8+调节性T细胞的作用机制

调节性T细胞（Treg）主要在机体免疫系统中发挥负向调节作用,既能抑制不恰当的免疫反应,又能限定免疫应答的范围、程度及作用时间,对CD4^＋和CD8^＋效应性T淋巴细胞的增殖起抑制作用,因此在移植物抗宿主病、自身免疫病、过敏性疾病等的发病机制和临床治疗中有潜在的应用价值.本文重点介绍CD4^＋CD25^＋Treg和CD8^＋Treg的作用机制,并简述调节性T细胞研究面临的挑战与展望.     

Differential increase of CD34, KDR/CD34, CD133/CD34 and CD117/CD34 positive cells in peripheral blood of patients with acute myocardial infarction

Objective Circulating progenitor cells (CPC) may contribute to cardiac regeneration and neovascularization after acute myocardial infarction (AMI). For potential therapeutic use, understanding the endogenous mechanisms after ischemia is inevitable. We investigated the absolute number, but also the subset composition of CD34+ CPC after AMI. Methods CD34+, KDR+/ CD34+, CD133+/CD34+ and CD117+/CD34+ CPC were analyzed by FACS in peripheral blood of 10 patients with acute MI (59±5 yrs, m/f=8/2) at day of AMI (day 0) and days 1–5. For comparison patients with stable coronary artery disease (CAD, n=12, 66±2 yrs, m/f=10/2) and young healthy volunteers (n=7, 26±2 yrs, m/f=3/4) were studied. Results CD34 and KDR/CD34, CD133/CD34, CD117/CD34 were increased day 3 and 4 after AMI. KDR+ fraction within CD34+ population remained unchanged (58.3±7.8% vs 55.3±10.6%), whereas CD133+ (64.9±3.1% vs 43.5±5.9%, P=0.006) and CD117+ fractions (71.7±5.6% vs 50.1±5.5%, P=0.02) were elevated. In CAD, all CPC and fractions were similar as AMI day 0. Healthy volunteers had more CD34+ than CAD and AMI day 0. Double positive CPC were also higher, but fractions were unchanged vs CAD with more KDR/CD34 in trend (72.8±10.6% vs 50.5±5.6%, P=0.058). After AMI both absolute numbers of CD34+ and their subset composition change, suggesting selective mobilization of CPC. Increased CPC after AMI never reach numbers of young healthy volunteers.     

CD28／CD152：CD80／CD86分子在强直性脊柱炎患者外周血淋巴细胞上的表达

目的探讨共刺激分子CD28／CD152：CD80／CD86在强直性脊柱炎（AS）患者外周血淋巴细胞上的表达及与免疫功能的关系。方法流式细胞仪检测AS患者及健康体检者T细胞表面标志CD3、CD4、CD8、CD19的表达及CD28／CD152和CD80／CD86在外周血T、B淋巴细胞上的表达水平；采用速率散射比浊法、酶联免疫吸附试验（ELISA）测定血清中免疫球蛋白IgG、IgA、IgM及C-反应蛋白（CRP）和白细胞介素-6（IL-6）水平。结果CD3^＋CD4^＋T细胞及CD4^＋／CD8^＋比值明显高于健康对照组（P〈0．05），而CD3^＋CD8^＋T细胞则明显低于健康对照组（P〈0．05）；CD4^＋和CD8^_T细胞上CD28和CD19^＋B细胞上CD80、CD86的表达均较健康对照组增高（P〈0．05），而CD152在CD4^＋T淋巴细胞上的表达也比健康对照组显著增加，但在CD8%＋T淋巴细胞上的表达却显著降低（P〈0.05）；血清IgG、IgA及CRP、IL-6水平均较健康对照组显著增高（P〈0．05）。结论AS患者外周血淋巴细胞CD28／CD152：CD80／CD86分子的异常表达与其免疫功能紊乱有密切关系。     

哮喘患儿CD8+CD28+、CD8+CD28-T淋巴细胞检测及其临床意义

目的通过对哮喘患儿CD4、CD8和CD28的联合检测,探讨哮喘患儿淋巴细胞免疫功能状态及其临床意义.方法采用流式细胞术检测哮喘患儿外周血的总T细胞(CD3+)、辅助/诱导T淋巴细胞(CD4+)、抑制/细胞毒T淋巴细胞(CD8+)、细胞毒T细胞(CD8+CD28+)、抑制T细胞(CD8+CD28-).结果哮喘患儿组与对照组比较;CD3+、CD4+、CD8+CD28-细胞均低于对照组(P＜0.01,P＜0.05),CD8+CD28+细胞增高(P＜0.05),CD4+/CD8+比值、CD8+与对照组比较均无显著性差异(P＞0.05).结论哮喘患儿存在T淋巴细胞亚群免疫功能紊乱,而CD8+CD28+、CD8+CD28-T细胞失衡可能是导致机体免疫功能紊乱的主要因素.     

Contribution of CD4+, CD8+CD28+, and CD8+CD28- T cells to CD3+ lymphocyte homeostasis during the natural course of HIV-1 infection.

The relationship between the number of circulating CD4+ T cells and the presence of particular CD8+ T cell subsets was analyzed by flow cytometry on PBL from asymptomatic HIV-1-infected patients whose specimens were collected every 2 mo for a total period of 32 mo. Only slight variations were detected in the absolute number of lymphocytes and percentage of CD3+ lymphocytes, whereas both CD4+ and CD8+ T cell subsets showed wide intrapatient variation. Variations in the number of CD8+CD28+ cells paralleled those of the CD4+ T cell subset in each patient tested, while the presence of CD8+CD28- T cells correlated inversely with CD4+ and CD8+CD28+ T cells. These data show that changes in the number of circulating CD4+-and CD8+CD28+ T cells are strongly related to the presence of CD8+CD28- T cells in these patients. Insight into the significance of CD8+CD28- T cell expansion will allow us to understand the mechanisms and significance of the HIV-1- driven change in CD4+CD8+ T cell homeostasis and the basic immunopathology of HIV disease.