慢乙肝急性发作后T细胞亚群变化及胸腺肽α1的调节作用

将32例HBc-IgM阳性慢性乙型肝炎(CHB)患者,分为HBc-IgM阳性对照组(常规保肝治疗),HBc-IgM阳性实验组(保肝及胸腺肽治疗)及HBc-IgM阴性对照组.治疗前后检测各组外周血CD 3、CD 4、CD 8T淋巴细胞百分比及CD 4/CD 8比值.结果与HBc-IgM阴性组比较,HBc-IgM阳性组CD 4 T细胞百分比升高;与HBc-IgM阴性组、HBc-IgM阳性实验组治疗前及HBc-IgM阳性对照组治疗后比较,HBc-IgM阳性实验组治疗后CD 4、CD 3 T细胞百分比及CD 4/CD 8 T细胞比值升高,CD 8T细胞百分比降低(P均<0.05).表明外周血CD 4T淋巴细胞在CHB急性发作后随着HBc-IgM转换而进行性下降,胸腺肽α1可逆转CD 4 T淋巴细胞的下降趋势.     

外周血T淋巴细胞亚群变化评估儿童慢性乙型肝炎预后转归

目的了解对干扰素有应答的儿童慢性乙型肝炎治疗前后与正常健康儿童外周血CD3+、CD3+CD4+及CD3+CD8+T淋巴细胞数量变化的差别,探讨儿童慢性乙型肝炎患者外周血T淋巴细胞亚群数量变化与临床预后的关系。方法对干扰素有应答的32例儿童慢性乙型肝炎患者抗病毒治疗前后及30例健康儿童进行外周血CD3+、CD3+CD4+及CD3+CD8+T淋巴细胞数量检测。结果儿童慢性乙型肝炎组(治疗前)外周血CD3+、CD3+CD4+及CD3+CD8+T淋巴细胞数量明显低于健康儿童组(P<0.01);治疗后患儿外周血CD3+、CD3+CD4+及CD3+CD8+T淋巴细胞数量明显高于治疗前(P<0.01)。结论儿童慢性乙型肝炎外周血CD3+、CD3+CD4+及CD3+CD8+T淋巴细胞数量变化可判断预后转归。     

高剂量乙肝疫苗对慢性乙型肝炎病毒携带者细胞免疫功能的影响

目的：观察高剂量乙肝疫苗对慢性乙型肝炎病毒（HBV）携带者细胞免疫功能的影响。方法：将82例慢性HBV携带者皮下注射乙肝疫苗60μg，每2周1次，连用3个月，使用流式细胞仪检测其治疗前后外周血T淋巴细胞亚群，PCR法检测HBV DNA定量，ELISA法检测γ-干扰素（IFN-γ）水平的改变。结果：慢性HBV携带者外周血T淋巴细胞CD3^＋基本正常，CD4^＋下降，CD8^＋上升；治疗组患者CD4^＋上升、CD8^＋下降、CD4^＋／CD8^＋上升，治疗前后比较差异有显著性意义（P〈0．05）；IFN-γ水平在治疗后明显升高，与治疗前比较差异有显著性意义（P〈0．05）。结论：高剂量乙肝疫苗能够提高慢性HBV携带者细胞免疫功能。     

不同临床类型HBV感染者外周血T淋巴细胞亚群的变化

目的研究HBV感染者外周血T淋巴细胞亚群的变化及其临床意义.方法在33例慢性乙型肝炎轻度、50例中度、32例重度、64例肝硬化、51例慢性乙型肝炎肝衰竭和30例健康人,采用特异性荧光抗体标记法和流式细胞仪检测外周血T淋巴细胞亚群.结果慢性乙型肝炎重度、乙型肝炎肝硬化和慢性乙型肝炎肝衰竭患者与正常对照组比,CD3^＋、CD4^＋、CD8^＋细胞数和CD4^＋/CD8^＋比值显著下降（P〈0.05）,以慢性乙型肝炎肝衰竭下降最显著;HBeAg阴性患者外周血CD3^＋、CD4^＋和CD8^＋细胞数较HBeAg阳性患者显著减少（P〈0.05）;HBV DNA载量与患者外周血CD3^＋、CD4^＋、CD8^＋细胞数和CD4^＋/CD8^＋比值均无明显相关性.结论严重的HBV感染者存在不同程度的细胞免疫功能低下及免疫调节紊乱,细胞免疫功能下降程度与病毒复制水平无明显相关.     

严重肝病患者并发感染性休克外周血T淋巴细胞亚群的分析

目的探讨并发感染性休克的肝病患者外周血T淋巴细胞亚群的变化。方法在42例并发感染性休克的慢性乙型肝炎患者,取外周血,采用特异性荧光抗体标记、BD流式细胞仪检测其T淋巴细胞亚群。结果慢性乙型肝炎并发感染性休克患者T淋巴细胞亚群表现为CD3 、CD4 、CD8 T淋巴细胞及CD4 /CD8 T比值明显下降(P<0.01),与无并发症组比较,CD3 、CD4 T淋巴细胞均明显下降(P<0.05),但CD8 T淋巴细胞无明显变化(P>0.05),导致CD4 /CD8 比值明显降低(P<0.05)。结论慢性乙型肝炎并发感染性休克患者T淋巴细胞亚群比无并发症组更紊乱,免疫功能更低下。     

安达芬对慢性乙型肝炎患者细胞免疫的影响

为探讨国产基因重组α-2b干扰素(安达芬)在调节慢性乙型肝炎患者细胞免疫功能的作用,以及细胞免疫功能与临床疗效的关系,我们观察了应用安达芬治疗的慢性乙型肝炎患者外周血T淋巴细胞亚群(CD3~+、CD4~+、CD8~+)、血清可溶性白细胞介素2受体(SIL-2R)、肿瘤坏死因子α(TNF-α)、乙型肝炎病毒标志物(HBVM),乙型肝炎病毒DNA(HBV DNA)治疗前后的动态变化。     

乙型肝炎患者外周血T淋巴细胞亚群的变化

目的研究乙型肝炎患者外周血T淋巴细胞亚群变化及其临床意义.方法对140例乙型肝炎患者和64例健康者,采用特异性荧光抗体标记、流式细胞仪检测T淋巴细胞亚群的改变.结果与正常对照组比较,慢性肝炎、肝炎肝硬变、慢性重型肝炎患者外周血CD3+、CD4+、CD8+及CD4+/CD8+比值均有显著性下降(P＜0.05),以肝炎肝硬变下降最显著,急性乙型肝炎T细胞亚群数量和比值变化不明显(P＞0.05).HBV DNA(+)与HBV DNA(-)患者外周血T细胞亚群数量和比值变化比较无显著性差异(P＞0.05).结论乙型肝炎病毒感染后机体T淋巴细胞功能低下主要见于慢性病毒感染状态,其下降程度与病毒复制水平无明显关系.     

慢性HBV感染及其相关肝病患者外周血T细胞亚群的变化

目的观察不同临床类型HBV感染者外周血T淋巴细胞亚群的差异,探讨HBV对人体T细胞免疫的影响及其可能机制,不同类型慢性HBV感染者免疫失衡的规律。方法用流式细胞仪技术检测患者外周血T细胞亚群。慢性HBV感染者318例,其中HBV携带者8例,慢性乙型肝炎231例,肝炎后肝硬化61例,原发性肝癌18例,观察患者的T淋巴细胞亚群、HBV DNA等。同时收集22名健康志愿者的新鲜血检测T淋巴细胞亚群。结果不同临床类型HBV感染者外周血CD3＋T、CD4＋T、CD8＋T细胞百分数低于正常对照组,差别均有统计学意义（P〈0.05或P〈0.01）,CD4＋/CD8＋比正常对照组低,但差异均无统计学意义（P〉0.05）。结论慢性HBV感染随着病情进展,由慢性肝炎→肝硬化→肝癌,直至细胞免疫功能逐渐衰退。慢性HBV感染的不同阶段的细胞免疫紊乱各具特点,就不同类型患者应采用不同的免疫调节治疗手段。     

恩替卡韦分散片对HBeAg阴性慢性乙型肝炎患者T细胞亚群的影响

目的探讨HBeAg阴性慢性乙型肝炎(CHB)患者接受恩替卡韦抗病毒治疗前后细胞免疫功能的变化。方法纳入HBeAg阴性CHB患者62例,接受恩替卡韦抗病毒治疗24周。通过流式细胞术检测患者治疗前后外周血T淋巴细胞亚群的变化。结果在治疗24周结束时,患者肝功能ALT、HBV DNA水平较治疗前明显下降,差异有统计学意义(P0.05);患者外周血CD3~+、CD4~+和CD8~+细胞计数较治疗前升高,CD4~+/CD8~+比值也显著高于治疗前,差异有统计学意义(P0.05)。结论恩替卡韦可能通过抑制HBV复制、降低CHB患者体内病毒载量,间接引起机体细胞免疫功能的改善。     

聚乙二醇干扰素对慢性丙型肝炎患者淋巴细胞亚群的影响

目的：研究聚乙二醇干扰素α-2b治疗慢性丙型肝炎（CHC）的疗效与机体免疫状态的关系。方法：采用流式细胞仪检测聚乙二醇干扰素α-2b治疗前后CHC患者T淋巴细胞亚群CD3＋、CD4＋、CD8＋T淋巴细胞及NK（自然杀伤细胞）、CTL（细胞毒性T淋巴细胞）的变化。结果：应答组患者血清CD3＋、CD4＋T淋巴细胞、NK细胞、CD4＋/CD8＋比值的上升程度及CD8＋、CTL细胞的下降程度明显高于无应答组,而持续应答组又相应优于复发组。结论：聚乙二醇干扰素α-2b治疗后患者的细胞免疫状态明显增强,检测患者T淋巴细胞亚群、NK、CTL细胞的变化,可以预测患者干扰素治疗的疗效。     

Effect of liniment levamisole on cellular immune functions of patients with chronic hepatitis B

AIM: To explore the effects of liniment levamisole on cellular immune functions of patients with chronic hepatitis B. METHODS: The levels of T lymphocyte subsets and mlL-2R in peripheral blood mononuclear cells (PBMCs) were measured by biotin-streptavidin (BSA) technique in patients with chronic hepatitis B before and after the treatment with liniment levamisole. RESULTS: After one course of treatment with liniment levamisole, the levels of CD3+, CD4+, and the ratio of CD4+/CD8+ increased as compared to those before the treatment but the level of CD8+ decreased. The total expression level of mIL-2R in PBMCs increased before and after the treatment with liniment levamisole. CONCLUSION: Liniment levamisole may reinforce cellular immune functions of patients with chronic hepatitis B.     

乙型肝炎病毒感染临床转归与宿主细胞免疫的相关性研究

目的 比较慢性乙型肝炎患者、HBV携带者、HBV既往无症状感染者之间T细胞亚群和HBV特异性CD4+ T细胞应答强度的差异,分析宿主的细胞免疫状态对HBV感染后临床转归的影响,探讨乙型肝炎的发病机制,为慢性乙型肝炎的治疗提供新的线索.方法 选取2004年2-10月在北京协和医院肝炎门诊就诊的慢性乙型肝炎患者30例、HBV携带者22例、HBV既往无症状感染者9例以及正常对照11例,使用流式细胞仪检测其T细胞亚群,使用酶联免疫斑点法检测病毒特异性CD4+T细胞应答强度,分析其差异及临床意义.结果 慢性乙型肝炎组CD4+T细胞计数显著低于HBV既往无症状感染组和正常对照组;慢性乙型肝炎组、HBV携带者组、HBV既往无症状感染组病毒特异性CD4+ T细胞应答强度分别为(156±105)、(56±68)、(229±114)SFC/106PBMC(每106个外周血单个核细胞中斑点形成细胞的个数);慢性乙型肝炎组明显高于HBV携带者组(P＜0.01),而低于HBV既往无症状感染组(P＜0.05).结论 慢性乙型肝炎患者、HBV携带者、HBV既往无症状感染者病毒特异性T细胞应答强度存在差异,这种差异可能是造成HBV感染后不同临床转归的主要因素之一.     

原发性肝癌和乙型肝炎肝硬化患者外周血淋巴细胞凋亡率、T细胞亚群和NK细胞的水平变化和临床意义

目的通过检测乙型肝炎肝硬化和合并乙型肝炎病毒感染的原发性肝细胞癌患者的外周血的粒细胞、单核细胞、NK细胞、T细胞及其亚群和淋巴细胞及其凋亡率,探讨两者在细胞免疫方面的差异.方法用Ficoll Hypaque离心法分离外周血单个核细胞(PBMNC),流式细胞仪检测外周血T细胞及其亚群、NK细胞和淋巴细胞、单核细胞和粒细胞,AnnexinV/FITCKit检测凋亡细胞.结果乙型肝炎肝硬化患者的外周血单核细胞、粒细胞、淋巴细胞、CD3-CD16 CD56 NK细胞、CD3 T细胞、CD3 CD4 T细胞、CD3 CD4 T细胞和CD4 CD8 T细胞比值,均与正常对照组无显著性差异(P＞0.05);但淋巴细胞凋亡率明显低于对照组(P＜0.01).原发性肝癌外周血CD3-CD16 CD56 NK细胞、单核细胞和CD4 /CD8 T细胞比值与肝硬化组和正常对照组比较,均无显著性差异(P＞0.05),而粒细胞明显升高(P＜0.05);CD3 T细胞、CD3 CD4 T细胞和CD3 CD8 T细胞均较另两组明显减少(P＜0.05),淋巴细胞及其凋亡率均明显低于另两组(P＜0.01).结论乙型肝炎肝硬化患者的外周血细胞免疫只发生不显著的变化,但淋巴细胞的凋亡率明显降低.原发性肝癌外周血的细胞免疫和淋巴细胞凋亡率均明显低下.     

Peripheral blood T,B, and NK cells in relation to histological hepatitis activity and fibrosis stage in chronic hepatitis C

BACKGROUND/AIMS: Many data on the pathogenesis of chronic hepatitis C have pointed to host's immune system disorders and a high variety of virus. However, there are no known criteria that could prognose the course of chronic hepatitis C infection. The analysis of T and B lymphocyte subpopulations in the peripheral blood was undertaken in patients with chronic hepatitis C of more than 6 months of duration. METHODOLOGY: Fluorescein isothiocyanate or phycoerythryne conjugated monoclonal antibodies for CD3+, CD4+, CD8+, CD19+, CD3++ HLA DR+, CD16++ CD56+ were used. The correlation between histological hepatitis activity and fibrosis (according Scheuer's scale) and the distribution of lymphocytes in the peripheral blood was sought. RESULTS: All patients with chronic hepatitis showed statistically significant increase in active lymphocytes CD3++ HLA DR+ and CD16++ CD56+ NK cells in peripheral blood. We observed the correlation between these cells and histological hepatitis activity and fibrosis. There was no correlation between the value of CD3+ and CD8+ cells and the stage of liver failure. In the early stage of chronic hepatitis C we noted decrease CD4+ cells with increase B cells CD19+. CD4+/CD8+ ratio was maintained as slightly decreased in chronic hepatitis C in favor of lymphocytes CD8+. CONCLUSIONS: The results show the correlation between peripheral blood value of activated T cell (HLA DR+) and NK cells with histological activity and fibrosis in chronic hepatitis C. Lymphocyte T (CD4+, CD8+) and B (CD19+) did not correlate with grade and stage of hepatitis C.     

The in vitro effect of interferon-alpha 2a on CD95 expression of T cells in hepatitis B

BACKGROUND/AIMS: In chronic hepatitis B, apoptotic rate of peripheral cytotoxic T cells may be related with hepatocyte injury. We aimed to investigate Fas (CD95) expression of peripheral cytotoxic T cells and to show the in vitro effect of interferon-alpha 2a on Fas expression and apoptosis. METHODOLOGY: The study group consisted of 17 patients with chronic hepatitis B and control group consisted of 10 healthy subjects. Apoptotic cells were identified by flow-cytometric assay using Annexin V and propidium iodide. We used monoclonal antibodies stained by direct immunofluorescent method to show surface molecules. CH-11 monoclonal antibodies were used as anti-Fas antibodies. RESULTS: Basal expressions of CD95 and CD8+ CD95+ in the peripheral mononuclear cells were higher in chronic hepatitis B patients than controls. In vitro treatment with interferon-alpha 2a increased the percentage of CD8+ 95+ peripheral mononuclear blood cells in controls; this effect was less remarkable in patients with chronic B hepatitis, and the difference was not statistically significant. The number of apoptotic cytotoxic T cells also increased in 5 subjects of the study group and 3 subjects of controls; the percentage of CD95+ cells increased in 7 of 17 patients and 8 of 10 controls. The percentage of CD8+ 95+ cells and the apoptotic rate of CD8+ cells were not different between study group and controls after combined treatment with interferon-alpha 2a and monoclonal stimulating anti-Fas antibodies. CONCLUSIONS: We suggest that interferon-alpha 2a does not induce Fas-dependent apoptosis in CD8+ peripheral T cells of the patients with chronic hepatitis B, and CD95 molecules on peripheral cytotoxic T cells might be defective.     

慢性乙型肝炎患者外周血CD8~+T淋巴细胞亚群T淋巴细胞受体β链互补决定区3谱型研究

目的 研究慢性乙型肝炎(CHB)患者外周血CD8+T淋巴细胞亚群T淋巴细胞受体(TCR)β链Ⅴ区(BV)中互补决定区3(CDR3)谱型特点.方法 采集8例CHB患者(ALT>2倍正常值上限)的抗凝血,分离外周血单个核细胞,磁珠分选CD8~+T淋巴细胞亚群,提取RNA并应用逆转录-聚合酶链反应扩增TCR BV中CDR3基因的24个家族,采用免疫指纹技术进行TCRBV各家族基因扫描和谱型分析.不同细胞亚群间数据比较采用配对t检验.结果 8例患者外周血淋巴细胞TCR BV家族CDR3谱型发生明显偏移,表现为单克隆性,寡克隆性及偏峰性克隆增生.8例患者CD8~+T淋巴细胞亚群发生TCR BV CDR3谱型偏移家族总个数高于去除CD8~+T淋巴细胞群的外周血单个核细胞[(10.6±4.7)个比(4.1±3.1)个,t=6.619,P<0.01)];比较单、寡克隆增生2种形式偏移的家族,CD8+T淋巴细胞亚群也高于代表CD4+T淋巴细胞的去除CD8~+T淋巴细胞群的外周血单个核细胞[(8.8±4.5)个比(3.9±2.8)个,t=5.706,P<0.01].对其中3例患者磁珠分选前后TCR BV谱型的比较发现,分选后CD8+T淋巴细胞亚群发生TCR BV谱型偏移家族数均高于分选前.结论 通过淋巴细胞亚群分选方式分析TCR BV家族CDR3谱型变化可以减少不同淋巴细胞亚群之间的峰型重叠干扰;应用这一方法分析外周血CD8~+T淋巴细胞亚群克隆增生程度有助于了解CHB患者炎症的发生机制.     

乙肝病人T细胞亚群、mIL-2R的变化及意义

目的检测乙型病毒性肝炎病人外周血T细胞亚群和膜白介素-2受体(mIL—2R)表达水平并探讨其在乙肝发病机制中的作用。方法采用生物素-链霉亲和素法对188例乙肝病人外周血单个核细胞(PBMC)进行T细胞亚群及植物血凝素(PHA)诱导前后mIL—2R水平测定。结果乙肝病人T淋巴细胞亚群中CD3~+、CD4~+降低,CD8~+升高,CD4~+/CD8~+比值降低,外周血单个核细胞mIL—2R表达水平在PHA诱导前后均降低,与正常对照相比,差异有显著意义(P<0.01);另外,乙肝病人PBMC中HBV—DNA(+)病人与PBMC内HBV—DNA(-)病人相比,mIL—2R表达水平亦存在较大差异(P<0.01)。结论乙肝病人体内存在明显的细胞免疫功能紊乱,T细胞活化障碍,并与肝病的慢性化相关。     

Immune formulation-assisted conventional therapy on anti-infective effectness of multidrug-resistant Mycobacterium tuberculosis infection mice

Objective:To study the effect of immune formulation-assisted conventional therapy on antiinfective ability of multidrug-resistant Mycobacterium tuberculous infection mice.Methods:BALB/c mice were used as experimental animals,multidrug-resistant Mycobacterium tuberculosis infection models were built,randomly divided into model group,moxifloxacin group,thymopentin group and combined treatment group and given corresponding drug intervention,and then colony numbers in the spleen and lung,T lymphocyte subset contents and programmed death-1(PD-1) expression levels in peripheral blood were detected.Results:Colony numbers in lung and spleen of moxifloxacin group and thymopentin group were significantly lower than those of model group and colony numbers in lung and spleen of combined treatment group were significantly lower than those of moxifloxacin group and thymopentin group:contents of CD3~+CD4~+T cells,Thl and Thl7 in peripheral blood of moxifloxacin group and thymopentin group were higher than dtose of model group,and contents of CD3~+CD8~+T cells.Th2 and Treg were lower than those of model group;contents of CD3~+CD4~+T cells.Th 1 and Th 17 in peripheral blood of combined treatment group were higher than those of moxifloxacin group and thymopentin group,and contents of CD3~+CD8~+T cells.Th2 and Treg were lower than those of moxifloxacin group and thymopentin group:PD-I expression levels on T lymphocyte,B lymphocyte and monocyte surface in peripheral blood of moxifloxacin group and thymopentin group were lower than those of model group,and PD-I expression levels on T lymphocyte.B lymphocyte and monocyte surface in peripheral blood of combined treatment group were lower than those of moxifloxacin group and thymopentin group.Conclusions:Immune formulation thymopentin can enhance the anti-infective ability of multidrug-resistant Mycobacterium tuberculosis infection mice,decrease bacterial load in lung and spleen,and enhance immune function.     

苦参碱对溃疡性结肠炎患者T细胞亚群的影响

[目的]探讨苦参碱注射液对溃疡性结肠炎（UC）患者外周血T细胞免疫的影响.[方法]采用免疫组化（SAP）法检测30例UC患者用苦参碱注射液治疗1月前、后和10例健康志愿者外周血T细胞亚群.[结果]30例UC患者中治愈7例,好转12例,无变化11例;UC患者CD8^＋Ts细胞明显减少;用苦参碱治疗后UC患者外周血CD3^＋、CD8^＋Ts细胞较治疗前明显增高,有效组CD8^＋Ts细胞显著高于无效组（P＜0.05）.[结论]UC患者抑制性T细胞减低,免疫亢进;用苦参碱注射液治疗有较好疗效,外周血T细胞,尤其CD8^＋Ts细胞水平明显增高.