T细胞异常活化在系统性红斑狼疮中的分子免疫机制

系统性红斑狼疮(SLE)是一种系统性自身免疫性疾病,其发病机制错综复杂。T细胞参与SLE自身免疫的启动和整个过程,且直接促进SLE受累器官的病理改变。其活化异常引起的细胞因子产生失衡、自身反应性T细胞持续存在、T细胞凋亡增加等,在SLE发病机制中发挥关键的作用。本文就近年来发现的SLE患者中T细胞活化在细胞膜信号,胞内激酶和转录因子等各个水平的异常及其对SLE发生发展的影响加以综述。     

The inhibition of human T cell proliferation by the caspase inhibitor z-VAD-FMK is mediated through oxidative stress

The caspase inhibitor benzyloxycarbony (Cbz)-l-Val-Ala-Asp (OMe)-fluoromethylketone (z-VAD-FMK) has recently been shown to inhibit T cell proliferation without blocking caspase-8 and caspase-3 activation in primary T cells. We showed in this study that z-VAD-FMK treatment leads to a decrease in intracellular glutathione (GSH) with a concomitant increase in reactive oxygen species (ROS) levels in activated T cells. The inhibition of anti-CD3-mediated T cell proliferation induced by z-VAD-FMK was abolished by the presence of low molecular weight thiols such as GSH, N-acetylcysteine (NAC) and l-cysteine, whereas d-cysteine which cannot be metabolised to GSH has no effect. These results suggest that the depletion of intracellular GSH is the underlying cause of z-VAD-FMK-mediated inhibition of T cell activation and proliferation. The presence of exogenous GSH also attenuated the inhibition of anti-CD3-induced CD25 and CD69 expression mediated by z-VAD-FMK. However, none of the low molecular weight thiols were able to restore the caspase-inhibitory properties of z-VAD-FMK in activated T cells where caspase-8 and caspase-3 remain activated and processed into their respective subunits in the presence of the caspase inhibitor. This suggests that the inhibition of T cell proliferation can be uncoupled from the caspase-inhibitory properties of z-VAD-FMK. Taken together, the immunosuppressive effects in primary T cells mediated by z-VAD-FMK are due to oxidative stress via the depletion of GSH.     

不同体外诱导物对T细胞疫苗诱导免疫耐受作用的影响

目的 :探讨不同的疫苗细胞体外诱导物对 T细胞疫苗 ( TVC)诱导免疫耐受作用的影响 ,方法 :用 L OU /C大鼠脾细胞免疫 BN大鼠 ,取免疫过的 BN大鼠脾细胞进行淋巴细胞转化实验 ,计算刺激指数 ( SI) ,比较 Con A和PHA两种诱导物对 T细胞的诱导增殖作用 ;用被 L OU/C大鼠抗原活化的 BN大鼠脾细胞作为疫苗细胞 ,在体外经Con A诱导增殖后灭活制备 TCV1、经 PH诱导增殖后灭活制备 TCV2 ,用制备的 T细胞疫苗分别免疫正常的 BN大鼠 ,于免疫后进行单向混合淋巴细胞反应 ( ML R) ,计算抑制率。结果 :淋巴细胞转化实验 :Con A的 SI为 2 1.4 2 ,显著高于 PHA的 SI( 4 .93) ( P<0 .0 1) ;ML R:TCV1免疫组抑制率为 87.75 % ,TCV2免疫组抑制率为 34.6 6 % ,与 TCV1免疫组比较 P<0 .0 1。结论 :用 Con A诱导的疫苗细胞制备的 T细胞疫苗 ,可以显著抑制被免疫大鼠脾细胞的免疫应答能力 ,体外诱导物的诱导增殖作用越大 ,所制备的 T细胞疫苗诱导免疫耐受的能力越强     

老年原发免疫性血小板减少症患者治疗前后T淋巴细胞亚群的临床研究

目的 检测老年原发免疫性血小板减少症(ITP)患者治疗前后T淋巴细胞亚群的动态变化, 探讨其在ITP发生发展中的作用。方法 采用流式细胞术测定ITP患者治疗前后及正常对照组外周血T淋巴细胞亚群的水平。结果 ITP患者治疗前后T淋巴细胞绝对值、CD3⁺、CD4⁺、CD8⁺、CD4⁺CD25⁺T淋巴细胞比例及CD4⁺/CD8⁺比值分别为(0.83±0.16)vs(1.74±0.36)、(71.71±1.07)% vs(72.69±1.35)%、(41.78±0.71)% vs(42.46±1.20)%、(29.67±0.97)% vs(28.56±1.75)%、(8.76±0.56)% vs(9.39±1.26)%、(1.42±0.07)vs(1.49±0.13), CD8⁺T淋巴细胞比例治疗后显著降低, 其余均显著升高, 差异有统计学意义(P<0.05)。结论 T淋巴细胞亚群的异常改变, 破坏自身免疫, 与病情相关, 可指导临床治疗, 并作为评估预后的参考指标。     

硫喷妥钠麻醉对小鼠T淋巴细胞亚群的影响

观察静脉麻醉药硫喷妥钠对小鼠Ｔ淋巴细胞亚群的影响,Ｔ淋巴细胞亚群测定应用单克隆抗体直接免疫荧光技术,由流式细胞仪测定,结果显示硫喷妥钠麻醉后２０ｈ小鼠外周ＣＤ４细胞,ＣＤ４／ＣＤ８比值显著下降,同时测定血中白细胞总数,实验组与对照组统计学处理无显著差异。表明硫喷妥钠麻醉可引起免疫抑制。     

奥沙利铂联合Exosome诱导的效应性T细胞抑制HepG2细胞增殖的研究

目的观察奥沙利铂(OXA)联合负载HepG2细胞裂解物的Exosome诱导的效应性T细胞对肝癌HepG2细胞系增殖的抑制作用。方法自健康人外周血中提取单个核细胞和T淋巴细胞,并以GM-CSF+IL-4的培养方案获得树突状细胞(DC)。将反复冻融后产生的HepG2细胞裂解物体外致敏DC,收集培养上清后以超高速离心法分离得到Exosome,电镜下观察其形态特征。用致敏DC及其分泌的Exosome刺激T淋巴细胞的增殖和活化。以ELISA试验检测不同刺激环境下T细胞分泌IL-1β、IL-2、IL-4、IL-6、IL-10、IFN-γ和TNF-α的能力。以细胞杀伤实验(MTT法)检测不同因素刺激后的T细胞、OXA(2.5、5.0、10.0mg.L-1)以及两者联合应用对HepG2细胞生长的抑制效应。结果与未致敏DC及其分泌的Exosome相比,HepG2细胞冻融裂解物致敏的DC及其分泌的Exosome能显著促进T细胞的增殖,并使其活化和分泌大量的IL-1β、IL-2、IL-6、IFN-γ和TFN-α等Th1型细胞因子。活化后的T细胞、OXA以及两者联合应用对HepG2细胞体外增殖均有抑制作用,而活化T细胞联合10mg.L-1OXA对HepG2细胞的杀伤率最高,但与联合5 mg.L-1OXA的效应相比,差异无统计学意义(P0.05)。结论负载肿瘤细胞抗原的DC所分泌的Exosome可在体外刺激T细胞,使其活化为具有抗肿瘤效应的细胞毒性T细胞,该细胞联合低浓度OXA能显著抑制肝癌细胞的体外增殖,并降低OXA的用量。     

调节性T细胞与炎性肠病研究进展

炎性肠病(Inflammatory bowel disease,IBD)为非特异性炎症性肠病,主要包括溃疡性结肠炎(Ul-cerative colitis,UC)和克罗恩病(Crohn’s disease,CD)。其病因和发病机制尚未完全明确。已知肠道粘膜免疫系统异常反应所导致的炎症反应在IBD中发挥着重要作用。笔者对调节性T细胞在肠道免疫系统中的作用与IBD的发病机制相关性的研究进展做一综述,为临床治疗IBD的新药研究提供可能性的途径。     

Prostaglandin E2 signaling through E prostanoid receptor 2 impairs proliferative response of double negative regulatory T cells

Limited data are available on the mechanisms that constrain the function of regulatory populations of T cells. Prostaglandin E2 (PGE2) is an endogenous membrane phospholipid metabolite that has important immunomodulatory effects on T cell function. Our previous microarray data indicated that E prostanoid receptor 2 (EP2), a receptor for PGE2, is expressed by regulatory αβTCR⁺ CD4⁻ CD8⁻ NK1.1⁻ double negative T (DN Treg) cell clones but not by their non-regulatory natural mutants. Hence, the hypothesis that PGE2 may influence DN Treg cell proliferation and/or regulatory function was tested in this study. Our data indicate that PGE2 acts via the EP2 receptor on DN Treg cells to inhibit their proliferation, an effect reproduced by the EP2-specific agonist butaprost and abrogated by the EP2 antagonist AH6809. In contrast, PGE2 did not affect the ability of DN Treg cells to kill syngeneic CD8⁺ T cells activated by allogeneic stimulation. Together, these findings suggest a role for PGE2 in limiting the expansion of DN Treg cells.     

Effects of lipopolysaccharide on T lymphocyte cell subsets and cytokine secretion in mesenteric lymph nodes of mice: Histological and molecular study

Lipololysaccharides (LPS) can disrupt the gut barrier. How dose LPS affects the immune performance of mesenteric lymph nodes? The results showed the hematological parameters significantly changed after LPS treatment. The length of intestinal villus was shortened and the depth of crypts was deepened, especially on the ileum. After LPS treatment 6 h, 12 h, the number of CD3⁺ T cells and CD4/CD8 in the mesenteric lymph nodes of ileum were reduced significantly; the levels of IFN-γ, TNF-ɑ and IL-2 were significantly decreased, and the levels of IL-6 and IL-10 were significantly increased in the ileum. The content of sIgA in the ileum was significantly decreased after LPS treatment 3 h, 6 h and was increased after LPS treatment 12 h. LPS through mesenteric lymph nodes, which induces the immune function reduced and the ileum injured obviously after treatment 6 h. Furthermore, the performance of intestinal immune performance was the lowest after LPS treatment 6 h.     

MNNG亚慢性暴露致大鼠食管损伤及炎性反应机制研究

目的 观察MNNG亚慢性暴露对大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)、白介素1β(IL-1β)含量及食管组织结构的影响.方法 65只SPF级雄性Wistar大鼠随机分为生理盐水对照组、80、120、160和200 μg/ml MNNG染毒组,每组13只.对照组大鼠采用等量生理盐水灌胃,实验组大...     

自然流产与T淋巴细胞及IL-2、IL-10的关系

目的 分析自然流产者外周血T淋巴细胞亚群(CD3 T、CD4 T、CD8 T细胞)及血清白细胞介素2(IL-2)、白细胞介素10(IL-10)的水平变化,探讨其在自然流产发病中的作用.方法 应用免疫荧光标记技术和流式细胞仪检测42例自然流产患者外周血T淋巴细胞亚群,ELISA法定量检测血清IL-2、IL-10,化学发光法检测血孕酮水平,并进行相关分析,同时与健康早孕组比较.结果 (1)自然流产者外周血CD3 T、CD4 T、CD8 T细胞水平明显高于健康早孕组(P＜0.05).细胞因子IL-2水平在自然流产组较健康早孕组明显升高,而IL-10和孕酮水平明显下降.(2)孕酮水平与细胞因子IL-2及CD4 T淋巴细胞呈显著负相关(P＜0.05),与IL-10呈正相关(P＜0.05).结论 细胞免疫异常可能与孕酮的减少相关,并可能是导致自然流产的重要原因之一.     

T细胞依赖性抗体反应检测方法研究进展

T细胞依赖性抗体反应(TDAR)是检测免疫功能的主要试验技术。根据使用抗原及抗原特异性抗体检测方法的不同,TDAR方法包括绵羊红细胞(SRBC)作为抗原的空斑形成细胞(PFC)方法、SRBC作为抗原的ELISA方法和钥孔戚血蓝素(KLH)作为抗原的ELISA方法。KLH作为抗原比SRBC稳定、易标准化且容易获得,ELISA检测方法操作简便、样本可保存,便于整合到临床前毒理学研究中。本文综述了TDAR检测方法对免疫毒性的预测作用及其研究进展。     

IL-2-PE40对免疫活性T细胞的影响

目的研究ＩＬ－２－ＰＥ４０对免疫活性Ｔ细胞的影响。方法采用ＣｏｎＡ刺激的淋巴细胞增殖试验、混合淋巴细胞培养（ＭＬＣ）及细胞毒试验。结果ＩＬ－２－ＰＥ４０对ＣｏｎＡ诱导的小鼠脾细胞有十分强的细胞毒性，能选择性地抑制ＭＬＣ中抗原活化的Ｔ细胞活性，保留未活化Ｔ细胞对ＣｏｎＡ诱导的丝裂原反应，在培养ｄ３加入ＩＬ－２－ＰＥ４０比培养开始时加入对ＭＬＣ抑制作用强。结论ＩＬ－２－ＰＥ４０能够高度选择性抑制免疫活性Ｔ细胞，是ＩＬ－２Ｒ靶向治疗中具有潜力的一种免疫抑制剂。     

轻度镇静对多发伤患者炎症反应的影响研究

目的 探讨ICU多发伤患者应用右美托咪定进行轻度镇静对炎症反应的影响.方法 选取在本院创伤外科ICU治疗的多发伤患者156例.采用随机数值表法将患者随机分为轻度持续镇静组(n=78)与常规镇静组(n=78).持续镇静组患者给予右美托咪定进行镇静治疗,常规镇静组患者随机接受咪达唑仑或丙泊酚治疗.收集患者临床资料,评估不同时间患者炎性指标的变化.结果 持续组患者机械通气时间显著低于常规组患者[(4.9±1.8)d比(8.8±2.0)d,P＜ 0.01],ICU住院时间显著短于常规组患者f(10.8±3.5)d比(16.8+4.2)d,P＜0.01].持续组患者治疗48 h后,患者血清IL-6、CRP与TNF-α水平显著低于常规组,差异均有统计学意义(均P＜0.05).而此时持续组患者血清IL-IO水平显著高于常规组(P＜ 0.05).患者治疗48 h后,血清CRP的药时曲线下表面积(AUG)最高,对患者发生脓毒症有一定预测价值.持续组与常规组病死率比较差异无统计学意义[8(10.3％)比11(14.1％),x2=0.539,P=0.462].结论 应用右美托咪定进行轻度持续镇静,可以降低炎症反应和炎症因子释放,改善患者预后,缩短ICU住院时间.     

乌司他丁减轻血流感染患者全身炎性反应的临床观察

目的 观察乌司他丁对减轻血流感染患者全身炎性反应的临床疗效.方法 选择重症监护病房（ICU）经病原学确诊为血流感染的患者27例,按随机原则分为试验组14例和对照组13例,2组给予相同的常规治疗,试验组在常规治疗的基础上加用乌司他丁.2组治疗时间均为7d.观察2组治疗前后急性生理学与慢性健康状况评分系统Ⅱ（APACHEⅡ）评分及C-反应蛋白（CRP）、降钙素原（PCT）水平.结果 2组治疗后APACHEⅡ评分、CRP、PCT水平较治疗前显著降低,且试验组低于对照组,差异有统计学意义（P〈0.05）.结论 乌司他丁能有效抑制血流感染患者的全身炎性反应,改善患者预后.     

间质干细胞对系统性红斑狼疮患者外周血T细胞亚群的影响

目的 观察间质干细胞（MSC）对系统性红斑狼疮（SLE）患者外周血T细胞亚群的影响。方法 从健康志愿者骨髓中分离培养MSC,从SLE患者外周血中分离单个核细胞（PBMC）,将MSC与混合淋巴细胞反应体系共培养。二乙酰羧基荧光素一琥珀酰亚胺酯（CFSE）标记反应细胞群结合流式细胞术特异性分析反应细胞各细胞亚群。结果 与MSC共培养后,SLE患者外周血T细胞增殖受抑制,各T细胞亚群比例降低,其中Th2亚群明显降低（P〈0．05）。结论 MSC对SLE患者体内Th1／Th2失衡状态有一定的纠正作用。     

T细胞接种对环磷酰胺诱导的NOD鼠淋巴细胞亚群的影响

目的　探讨 T细胞接种 (TCV)对 1型糖尿病预防作用的机制。方法　 4周龄未发病的雌性 NOD鼠用 18周龄新近发病的 NOD鼠脾细胞制得的 T细胞疫苗接种 ,检测 TCV对环磷酰胺诱导的 NOD鼠淋巴细胞亚群的变化情况。结果　 TCV可以诱导宿主脾脏 CD8+ T细胞百分比升高、CD4+ / CD8+比值和 IL- 2 R+细胞下降 ,以及胸腺 CD4+、CD8+单阳性细胞百分比升高。结论　 TCV可以诱导宿主产生自身免疫低反应性 ,这种作用可能与宿主脾脏和胸腺淋巴细胞亚群的变化有关。     

初治肺结核患者T淋巴细胞亚群和Th细胞的变化及与预后的关系

目的：探讨初治肺结核患者T淋巴细胞亚群和Th细胞的变化及与预后的关系。方法选取2011年1月～2013年6月在本院结核科门诊治疗的肺结核患者60例为肺结核组,按初治疗效进一步将肺结核组分为好转组与未好转组,选取健康未患肺结核30例为对照组。测定外周血T细胞亚群、Th细胞及IFN-γ、IL-4。结果治疗前,未好转组CD4^＋、CD4^＋/CD8^＋、Th1/Th2、Th1表达较对照组与好转组显著降低,Th2表达较对照组与好转组显著升高（P〈0.05）;治疗前,好转组CD4^＋、Th1/Th2、Th1表达较对照组低,Th2表达较对照组高（P〈0.05）;治疗后,好转组CD4^＋、CD4^＋/CD8^＋、Th1/Th2、Th1显著高于未好转组及治疗前,Th2显著低于未好转组及治疗前,差异有统计学意义（P〈0.05）;与治疗前比较,未好转组治疗后CD4^＋、CD8^＋、CD4^＋/CD8^＋、Th1/Th2、Th1、Th2差异无统计学意义（P〉0.05）。结论初治肺结核患者 CD4^＋、CD4^＋/CD8^＋、Th1/Th2、Th1较正常人降低,Th2较正常人升高;肺结核治疗初期,CD4^＋、CD4^＋/CD8^＋、Th1/Th2、Th1偏高,Th2偏低的患者预后较好;治疗有效后CD4^＋、CD4^＋/CD8^＋、Th1/Th2、Th1升高,Th2下降。     

Fasudil regulates T cell responses through polarization of BV-2 cells in mice experimental autoimmune encephalomyelitis

Aim:

Fasudil, a selective Rho kinase (ROCK) inhibitor, has been shown to alleviate the severity of experimental autoimmune encephalomyelitis (EAE) via attenuating demyelination and neuroinflammation. The aim of this study was to investigate the effects of fasudil on interactions between macrophages/microglia and T cells in a mice EAE model.

Methods:

Mouse BV-2 microglia were treated with IFN-γ and fasudil. Cell viability was detected with MTT assay. BV-2 microglia polarization was analyzed using flow cytometry. Cytokines and other proteins were detected with ELISA and Western blotting, respectively. Mice were immunized with MOG_35–55 to induce EAE, and then treated with fasudil (40 mg/kg, ip) every other day from d 3 to d 27 pi. Encephalomyelitic T cells were prepared from the spleen of mice immunized with MOG_35–55 on d 9 pi.

Results:

Treatment of mouse BV-2 microglia with fasudil (15 μg/mL) induced significant phenotype polarization and functional plasticity, shifting M1 to M2 polarization. When co-cultured with the encephalomyelitic T cells, fasudil-treated BV-2 microglia significantly inhibited the proliferation of antigen-reactive T cells, and down-regulated IL-17-expressing CD4⁺ T cells and IL-17 production. Furthermore, fasudil-treated BV-2 microglia significantly up-regulated CD4⁺CD25^high and CD4⁺IL-10⁺ regulatory T cells (Tregs) and IL-10 production, suggesting that the encephalomyelitic T cells had converted to Tregs. In EAE mice, fasudil administration significantly decreased both CD11b⁺iNOS⁺ and CD11b⁺TNF-α⁺ M1 microglia, and increased CD11b⁺IL-10⁺ M2 microglia.

Conclusion:

Fasudil polarizes BV-2 microglia into M2 cells, which convert the encephalomyelitic T cells into Tregs in the mice EAE model.     

急性肾炎患儿治疗前后B细胞和T细胞亚群的检测

目的　探讨急性肾炎患儿治疗前后外周血 B细胞和 T细胞亚群含量。方法　应用单克隆抗体技术测定 30例小儿急性肾炎外周血 B细胞和 T细胞亚群 ,并以 35名健康人作对照。结果　急性肾炎患儿外周血 B细胞数显著地高于正常对照组 (P<0 .0 1) ,CD3、CD4、CD4/CD8显著地低于正常人对照组 (P<0 .0 1或 P<0 .0 0 1)。结论　急性肾炎患儿为一种自身免疫调节异常的疾病。