丙型肝炎患者外周血T淋巴细胞亚群的变化

丙型肝炎患者外周血Ｔ淋巴细胞亚群的变化李灼亮，郑茉莉，谢庆，涂荫国，黄振国空军广州医院传染病科广州５１０６０２一般认为［１～４］慢性乙型肝炎患者外周血Ｔ淋巴细胞亚群的变化，可反映患者全身免疫状态，ＣＤ８＋细胞显著升高，ＣＤ４＋／ＣＤ８＋比值下降，可能...     

梅毒病人外周血T细胞亚群特点的探讨

为了解被梅毒螺旋体感染后病人的细胞免疫功能的变异情况，对４０例梅毒病人进行了Ｔ细胞亚群的检测。先用ＴＲＵＳＴ筛选出阳性者，以再生ＴＰ，ＰＡ确诊试验的阳性者为对象，与正常组进行比较。用ｔ检验进行统计学处理。阳性组，Ⅱ组ＣＤ３，ＣＤＥ４无显著差异，ＣＤ８显著高于对照组，ＣＤ４／ＣＤ８比值显著低于对照组。     

结缔组织疾病患者外周血T细胞亚群测定

<正> 本文采用人T细胞单克隆抗体OKT系列的直接血球测定法,检测了类风湿性关节炎(RA)及全身性红斑狼疮(SLE)患者共45例,以了解该病治疗前后,T细胞亚群的变化。     

带状疱疹患者外周血T淋巴细胞亚群分析

带状疱疹患者外周血Ｔ淋巴细胞亚群分析李大宁，余红上海第二医科大学附属新华医院皮肤科上海２０００９２带状疤疹是一种由水痘一带状疱疹病毒（ＶＺＶ）感染引起的疾病，机体对ＶＺＶ的特异性免疫目前认为主要是细胞免疫。本文采用流式细胞仪（ＦＣＭ）技术测定带状疤疹...     

乳癌患者外周血及肿瘤局部单个核细胞亚群分析

对13例女性乳癌患者外周血MNC(半个核细胞)与15名正常女性比较时发现,乳癌患者血中Leul~+细胞及DR~+细胞较低(P<0.05,P<0.01),Leu2a~+细胞较高(P<0.01),Leu3a/Leu2a比值下降(P<0.001),从数量上反映患者机体细胞免疫功能紊乱,并发现可能存在的Leu3a~+、Leu2a~+双标记细胞增多的现象,也可能与免疫抑制有关。DR~+细胞的减少由何引起有待于进一步研究。配对资料分析表明:癌周间质MNC亚群与血中MNC各亚群水平相近(Leu7、Leullb细胞除外),而癌间间质则显示较低的MNC水平。     

外周血T淋巴细胞亚群检测在丙型肝炎干扰素治疗中的作用

外周血Ｔ淋巴细胞亚群检测在丙型肝炎干扰素治疗中的作用孔丽刘金星曹治宸李兵顺（河北医科大学第三医院传染科，石家庄０５００５１）作者简介：孔丽，女，３３岁，主治医师，主要从事病毒性肝炎研究；刘金星，男，５３岁，主治医师，主要从事病毒性肝炎研究干扰素（ＩＦ...     

海洛因依赖者外周血T细胞亚群观察

数年前著者曾观察过海洛因依赖者外周血中CD3 、CD4 、CD8 细胞及CD4 /CD8 比值,现报告如下。1　对象及方法30例调查对象来自昆明4所康复医院或戒毒所。年龄14～40岁(多为年青者),其中男23例,女7例。全部系静脉注射“4号海洛因”(纯度>85%)成瘾者,平均日用量0.4～1.9g,平均吸毒时间19.6个月,符合海洛因依赖之条件;HIV血清学检测阴性反应,并排除慢性肝病、免疫性及内分泌性等疾病。对照组为青年健康者,男20例,女15例。CD系列MCAb试剂系武汉生物制品研究所产品。方法为IFA。资料经t检验。2　结果海洛因依赖者CD 细胞的表达及其与正…     

吗啡依赖小鼠胸腺,外周血T淋巴细胞亚群动态研究

使用流式细胞仪（ＦＣＭ）动态观察吗啡依赖小鼠在摄入吗啡成瘾期间及吗啡消除后中枢和外周血Ｔ淋巴细胞亚群的变化情况。结果表明：①摄入吗啡后２４ｈ始，胸腺中ＣＤ４和ＣＤ８Ｔ淋巴细胞降低，血液中ＣＤ４降低，ＣＤ８升高，两者比值倒置，连续摄药至ｄ５时最甚。②停止摄入吗啡７２ｈ后，胸腺重量增高，胸腺中ＣＤ４和ＣＤ８Ｔ淋巴细胞即恢复至正常；但吗啡消除１ｗｋ后，血液中的ＣＤ４和ＣＤ８Ｔ淋巴细胞始终未恢复正常。上述结果提示中枢和外周的淋巴细胞亚群对吗啡的反应敏感性存在差异，还提示吗啡在体内对中枢未分化成熟Ｔ淋巴细胞的可逆性损伤或抑制作用和对成熟Ｔ细胞的损伤或抑制更为严重，且有不可逆的作用     

白血病患者外周血T淋巴细胞亚群和SOD水平

据文献报告[1] ,超氧化物歧化酶与肿瘤的关系颇受临床的关注。而有关白血病患者ＳＯＤ水平与Ｔ淋巴细胞亚群水平的报道不多 ,为此我们进行了探讨 ,旨在了解白血病患者的免疫功能的变化 ,现报道如下。对象和方法一、对象 :(一 )正常人 :35人。均为我院预防保健科体检合格的健康人 ,无心、肝、肺、肾等重要脏器疾患 ,肝、肾功能试验正常 ,无血液病史。(二 )病人组 :32人。均为临床、血常规、骨髓像、常规组织化学染色检查确诊。其中急性非淋巴细胞性白血病 12例 ,Ｍ14例、Ｍ2 6例、Ｍ52例 ,急性淋巴细胞白血病 2 0例 ,Ｌ16例、Ｌ2 9例、Ｌ35例…     

EB病毒转化人外周血单个核细胞作为HCV复制模型的初步研究

目的　复制丙型肝炎病毒（ＨＣＶ） 感染的细胞模型。方法　用ＥＢ病毒感染其ＨＣＶ 阳性患者的外周血单个核细胞并使其转化，获得ＨＣＶ的外周永生化Ｂ细胞株。以后，每隔１ 个月应用套式逆转录聚合酶链反应（ＲＴＰＣＲ） 检测１ 次培养细胞和上清中的ＨＣＶＲＮＡ。结果　ＨＣＶ可以在传代细胞中持续存在１ 年，培养细胞中的ＨＣＶ负链和培养上清中的ＨＣＶ 则呈间断阳性。结论　ＨＣＶ 可以在该细胞株中较长时间存在、复制和分泌     

Quantitation of hepatitis C virus in liver and peripheral blood mononuclear cells from patients with chronic hepatitis C virus infection

Since the natural history of hepatitis C virus-associated liver disease and the therapeutic responsiveness might vary according to liver and blood mononuclear cells viral levels, it may be important to quantitate viral RNA in liver, blood mononuclear cells and serum, and to compare these data with genotype, biochemical and histologic data. A polymerase chain reaction-based assay available for serum hepatitis C virus RNA quantitation has been optimized to quantitate viral genomes in liver and peripheral blood mononuclear cells from 47 chronic hepatitis C patients. The procedure permitted hepatitis C virus RNA quantitation in freshly isolated mononuclear cells and in total RNA extracted from frozen mononuclear cells and liver tissue. The intrahepatic viral amount (median: 2.6 × 10³ copies/μg RNA; range: 0 to 3.6 × 10⁴ copies/μg RNA) correlated significantly with the hepatitis C virus RNA concentration in serum (r = 0.76, P < .001) but not in mononuclear cells. Viral RNA concentrations in liver (P < .001), serum (P < 0.01) and PBMC (P < 0.05) were significantly higher in hepatitis C virus genotype 1 patients (essentially type 1b) than in non-1 type cases, but were unrelated to biochemical or histologic indexes of disease activity. In conclusion, the optimized assay permit HCV RNA quantitation in liver and peripheral blood mononuclear cells, suggesting that serum viral level is an accurate measurement of intrahepatic viral burden. J. Med. Virol. 54:265–270, 1998. © 1998 Wiley-Liss, Inc.     

丙型肝炎病毒核心蛋白在患者外周血单个核细胞内的核表达检测

目的　检测和研究丙型肝炎病毒 (hepatitisCvirus,HCV)核心蛋白在患者外周血单个核细胞 (peripheralbloodmononuclearcells ,PBMC)内核表达的意义 ,并探讨其与临床的关系。方法　对 6 6例慢性丙型肝炎患者PBMC标本进行免疫组化检测 ,并将HCV蛋白抗原定位分布情况与患者临床状况进行比较分析 ,对其中 2 7例患者PBMC进行HCVRNA和HCVAg的平行检测和分析。结果　免疫组化结果显示 ,慢性丙型肝炎患者PBMCHCVAg(core +NS3)阳性检出率为 77 2 7% (5 1 6 6 )。结果还证实 ,HCV核心蛋白均定位于胞核内 ,且呈强表达 ;NS3蛋白主要定位于胞质内 ,呈弱表达。当进行HCVAg在PBMC内定位情况与患者临床状况比较分析时显示 ,病情较重患者PBMC内核心蛋白表达阳性率 (35 2 9% )明显高于病情较轻者 (5 88% ) (P <0 0 0 1)。结论　HCV核心蛋白在PBMC内核表达与患者临床状况相关 ,提示其可能是丙型肝炎慢性化的一个指标 ,并可能在肝硬化和肝癌发生上起一定作用     

Prominent proliferative response of peripheral blood mononuclear cells to a recombinant non-structural (NS3) protein of hepatitis C virus in patients with chronic hepatitis C.

The proliferative response of peripheral blood mononuclear cells (PBMC) to a recombinant non-structural (NS3) protein of hepatitis C virus (HCV) was studied in 41 patients with chronic hepatitis C. Of them, 28 had chronic persistent hepatitis (CPH) and 13 chronic active hepatitis (CAH). The positive proliferation rate of PBMC to the recombinant NS3 protein, T9Ag, was 66% in the 41 patients (77% in CAH versus 61% in CPH; P > 0.05) when stimulation index (SI) = 4 was set as the cut-off value. However, mean SI of CAH patients was significantly higher than that of CPH patients (8.3 +/- 5.2 versus 5.1 +/- 3.6; P < 0.05). Six other chronic hepatitis patients who were repeatedly negative for anti-HCV antibody but positive for serum HCV RNA also had an SI of > or = 4.0. The frequency of cellular immune response to the T9Ag is among the highest results obtained by using HCV antigens tested so far. Our studies thus indicate that NS3 is an immunologically important region of HCV for T cells. Moreover, the proliferative response to T9Ag may help to establish hepatitis C etiology in chronic hepatitis patients who are seronegative with currently available anti-HCV assays.     

羊水来源间充质细胞对于外周单核淋巴细胞增殖抑制作用的机制研究

目的鉴定羊水来源间充质细胞(AF-MCs)并探讨其对于外周单核淋巴细胞(PBMC)增殖活性的影响及机制。方法通过选择性消化和单克隆培养分离及培养人源羊水细胞(AFCs),而后通过核型分析细胞来源和流式分析细胞表型鉴定其为间充质细胞(MCs)。另一方面,将鉴定得到的AF-MCs与PBMC共培养,检测其是否具有免疫抑制功能。运用流式分析CD4~+CD25~+Foxp3~+T细胞频率在共培养前后变化,探讨AF-MCs具有免疫抑制功能的机制。结果AF-MCs来自于胚胎发育过程中的脱落细胞,其具有与骨髓间充质干细胞(BM-MSCs)相似的表型,且其能上调CD4~+CD25~+Foxp3~+T细胞的频率而产生免疫抑制功能。结论AF-MCs具有与BM-MSCs类似的表型,并且在免疫调节方面与BM-MSCs类似。这为临床应用间充质干细胞提供了一个新的,安全的种子细胞来源。     

Optimisation of herpes simplex virus-based vectors for delivery to human peripheral blood mononuclear cells

Peripheral blood mononuclear cells (PBMCs) represent a significant target for gene delivery both for therapeutic and experimental purposes. Thus far however, it has proved difficult to develop vectors capable of high efficient gene delivery to unstimulated PBMCs. We have tested a range of different vectors derived from herpes simplex virus (HSV) which differ in their degree of disablement in terms of their gene delivery efficiency to unstimulated human PBMCs and ability to deliver a reporter gene. None of the viruses had any significant toxic effect in PBMCs. However, optimal gene delivery to unstimulated PBMCs was obtained with a semidisabled virus lacking functional genes encoding ICP34.5 and Vmw65 which was more efficient than either nondisabled or more extremely disabled viruses. Expression of green fluorescent protein (GFP) with this virus was observed in up to 50% of PBMCs 1 day after infection, and reporter gene expression was detectable by Western blotting and immunofluorescence at undiminished levels at the longest time points tested, up to 5 days after infection. This optimised HSV vector may thus represent an effective tool for gene delivery to unstimulated PBMCs in culture.     

Quantitation of hepatitis C virus RNA in peripheral blood mononuclear cells in HCV-monoinfection and HIV/HCV-coinfection

Peripheral blood mononuclear cells (PBMCs) represent an extrahepatic hepatitis C virus (HCV) reservoir, the significance of which is unclear due to limited studies and varying test methodologies. In this study, a commercial viral load assay for measuring cell‐associated PBMC HCV RNA was evaluated. HCV RNA was extracted from PBMCs, sorted CD14+, and CD19+ cells and corresponding plasma samples using the Abbott m2000 and Real‐Time HCV assay. Test performance and influence of HIV seropositivity on plasma and PBMC HCV RNA were studied. Among 51 patients, 67 and 62 unique patient samples had detectable plasma and PBMC HCV viral load, respectively. The median PBMC viral load was 535 IU/1 M cells (range 29–5,190). CD19+ cells had significantly higher viral load than CD14+ cells (median log₁₀ HCV viral load 2.63 vs. 1.50 IU/ml; P< 0.001). Stability of PBMC viral load over time was demonstrated in untreated patients; all patients with an undetectable plasma HCV viral load after HCV treatment also demonstrated undetectable PBMC viral load. Repeated testing in nine samples yielded consistent PBMC viral load, differing by only 1.3‐fold (range 1.0–1.7‐fold). Among samples with detectable plasma HCV RNA, the correlation between PBMC and plasma viral load was moderate (r = 0.66) and was greater among HCV mono‐infected compared to HIV/HCV co‐infected subjects (r = 0.80 vs. 0.52). Measurement of cell‐associated PBMC HCV RNA using a commercial assay demonstrated promising test characteristics. Differences in PBMC HCV viral load based on HIV‐coinfection status and the significance of greater copy number in B‐cells requires further study. J. Med. Virol. 84:431–437, 2012. © 2011 Wiley Periodicals, Inc.     

慢性丙型肝炎患者抗病毒治疗前PBMC细胞因子的研究

目的 研究准备抗病毒治疗的慢性丙型肝炎患者的免疫特点.方法 将30例慢性丙型肝炎患者和10例正常对照外周血单个核细胞(PBMC)体外培养72 h后,用ELISA法检测培养上清中细胞因子IL-2、IL-4、IL-10、IL-12、IFN-γ和TNF-α的浓度.结果 (1)慢性丙型肝炎患者PBMC培养上清中IFN-γ、IL-10和TNF-α的水平明显升高(P＜0.05),没有检测到IL-2、IL-4、IL-12的基础分泌.(2)不同病情患者间的细胞因子的分泌水平差异无统计学意义(P＞0.05).结论 慢性丙型肝炎患者体内TH2型细胞因子的分泌占优势.提示通过调整TH1/TH2失衡可能达到抗病毒治疗目的.     

C-reactive protein is expressed and secreted by peripheral blood mononuclear cells

C-reactive protein (CRP) protects against bacterial pathogens and is a predictor of cardiovascular events. CRP is produced by vascular and organ-specific cells but the generation of CRP from peripheral blood mononuclear cells (PBMC) is poorly established. In a randomized, double-blind, placebo-controlled, two-way cross-over trial six healthy volunteers received a bolus infusion of 20 IU/kg Escherichia coli endotoxin [lipopolysaccharide (LPS)] or placebo. Intracellular CRP protein and CRP secretion of peripheral blood mononuclear cells (PBMC) was measured at baseline and 6 h after LPS by flow cytometry and enzyme-linked immubosorbent assay (ELISA), respectively. CRP mRNA expression was determined by real-time polymerase chain reaction (PCR). Regulation of the expression pathway was assessed using specific inhibitors in vitro. Small amounts of CRP protein and mRNA were detectable in PBMC, which were up-regulated between two- and eightfold by endotoxaemia in vivo. Augmented expression and release of CRP by LPS was consistent in PBMC cell culture experiments. LPS, interleukin (IL)-1, IL-6 and tumour necrosis factor (TNF)-alpha increased and IL-10 reduced CRP expression in PBMC. Toll-like receptor (TLR)-4, nuclear factor (NF)-kappaB and protein kinase C (PKC) activation were identified as intracellular signal transduction pathways of LPS-induced CRP expression. Constitutive CRP expression and release in PBMC is enhanced by inflammatory stimuli in vivo and in vitro. LPS might induce CRP generation via activation of TLR-4, NF-kappaB and PKC.