HIV感染者CD4~+T细胞TIGIT受体及其配体CD155表达情况研究

目的研究人类免疫缺陷病毒(human immunodeficiency virus,HIV)未治疗感染者CD4~+T细胞表面TIGIT受体及其配体CD155表达的情况。方法选取24例未经高效抗逆转录病毒治疗的HIV感染者和20例HIV抗体阴性健康对照(HIVnegative normal control,NC),用流式细胞仪检测受试对象外周血CD4~+T细胞表面TIGIT受体及其相关配体CD155的表达情况。结果未治疗HIV感染者CD4~+T细胞TIGIT受体表达百分数较健康人组表达明显增高(P0.000 1),并且与CD4~+T细胞绝对计数呈负相关(r=-0.444 1,P=0.029 7)。在未治疗HIV感染者中,CD4~+T细胞计数350/μl组CD4~+T细胞TIGIT表达百分数明显低于CD4~+T细胞计数≤350/μl组(P=0.029 2);病毒载量10~5/ml组CD4~+T细胞TIGIT表达百分数明显高于病毒载量≤10~5/ml组(P=0.015 5)。未治疗HIV感染者CD4~+T细胞表面CD155表达百分数与健康人组相比明显增高(P=0.004 2),且与病毒载量呈正相关(r=0.467 7,P=0.021 2),其中病毒载量10~5/ml组CD4~+T细胞CD155表达百分数明显高于病毒载量≤10~5/ml组(P0.000 1)。结论未治疗HIV感染者CD4~+T细胞TIGIT受体及其配体CD155表达百分数明显升高,且与CD+4绝对计数及病毒载量存在关联性,可为HIV患者的治疗和预后评估提供重要科学依据。     

无症状期HIV感染者CD4+和CD8+T细胞计数与病毒载量的关系

艾滋病是由人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染所导致的以免疫系统功能缺陷为特征的慢性高致死率传染病.CD4+T细胞是HIV损害的主要靶细胞,HIV可引起CD4+T淋巴细胞进行性丢失与功能受限[1-2].在HIV感染早期,CD8+T细胞增多并通过分泌各种细胞因子杀死被病毒感染的靶细胞,其高表达与病毒量载量呈正相关[3],而随着疾病进展,CD8+T细胞会消耗性减少.CD4+T细胞、CD8+T细胞计数与病毒载量的相关性一直为研究者关注[4-6].本研究检测了无症状期感染者,外周血CD4+T细胞和CD8+T细胞计数与病毒载量并分析它们的相关性.     

免疫调控受体PD-1和TIGIT在HIV感染者/HIV患者CD3~+CD4~-T细胞表达研究

深入了解免疫调控受体程序性死亡分子1(programmed death 1,PD-1)和T细胞免疫球蛋白及免疫酪氨酸样抑制基序(T cell immunoglobulin and ITIM domain,TIGIT)在人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染者/HIV患者外周血CD3~+CD4~-T细胞的表达情况及其与疾病进展的关系。用流式细胞仪检测24例未治疗的HIV感染者/HIV患者外周血CD3~+CD4~-T细胞表面PD-1和TIGIT的表达百分比、CD4+T细胞绝对值计数,实时荧光定量PCR检测HIV病毒载量,并与20例HIV抗体阴性者表达情况比较(HIV-negative normal control,NC)。结果发现HIV感染者/HIV患者CD3~+CD4~-T细胞表面TIGIT受体表达百分数较健康对照组明显增高(P0.000 1),与CD4+T细胞绝对计数呈负相关(r=-0.450 4,P=0.027 1),与病毒载量呈正相关(r=0.621 4,P=0.001 2);HIV感染者CD3~+CD4~-T细胞PD-1受体表达百分数较健康对照组明显增高(P0.000 1),与CD4+T细胞绝对计数呈负相关(r=-0.455,P=0.0255);PD-1和TIGIT共表达的CD3~+CD4~-T细胞百分数在HIV感染者/HIV患者中表达明显增高(P0.000 1)。HIV感染机体后,CD3~+CD4~-T细胞PD-1受体和TIGIT受体表达百分数均明显增高,且与HIV疾病进展密切相关。     

HIV/AIDS患者疾病进程与CD4+CD25hi调节性T细胞之间相关性研究

目的 通过分析不同阶段HIV感染者外周血CD4+CD25hi调节性T细胞(CD4+CD25hiregulatory T cells,Treg cells)与外周血免疫状态和病毒载量的相关性,探讨Treg细胞对HIV/AIDS发病进程的影响.方法 采集116例HIV感染者和21例正常人对照外周血,用4色流式细胞术进行CD4+和CD8+T细胞绝对数计数;用3色流式细胞术进行Treg细胞测定;用荧光定量PCR法进行HIVRNA载晕测定.实验数据用回归统计学方法和T检验方法进行分析.结果 HIV感染者外周血Treg细胞频率在HIV感染初期显著下降,之后随着疾病的进程逐渐升高.在CD4+T细胞大于300/μl的患者低于正常对照组,在CD4+T细胞小于100/μl的患者高于正常对照组,差异具有统计学意义.Treg细胞频率与CD4+T淋巴细胞绝对数和CD4+/CD8+之间均呈负相关.其相关系数r和P值各为r=-0.564,P＜0.001和r=-0.377,P＜0.001;Treg细胞频率与血浆HIV病毒载量呈正相关,其相关系数r=0.514.P＜0.001.结论 CD4+CD25hi Treg细胞可能是参与艾滋病免疫发病机理的重要细胞,在HIV感染发病进程的不同阶段具有不同的意义,其确切机制有待进行进一步研究.     

中国HIV感染长期不进展者CD4+CD25+Foxp3+调节性T细胞变化研究

目的 对中国HIV感染长期不进展者(LTNP)CD4+CD25+Foxp3+调节性T细胞水平及其与疾病进展相关性进行研究,探讨CD4+CD25+Foxp3+ 调节性T细胞在LTNP保护机制中发挥的作用.方法 选取74名HIV-1感染者(LTNP、典型进展HIV组、AIDS组)及16名健康对照,应用流式细胞仪胞内染色技术在单细胞水平检测CD4+CD25+Foxp3+调节性T细胞表达水平,分析其与CD4+ T细胞数量、病毒载量、淋巴细胞活化、凋亡水平的相关性.结果 中国HIV感染LTNP CD4+CD25+Foxp3+ T细胞百分率明显低于典型进展HIV、AIDS组及健康对照组(P<0.05).HIV/AIDS患者CD4+CD25+Foxp3+ T细胞百分率与CD4+ T细胞显著负相关(r=-0.509,P<0.001),与病毒载量明显正相关(r=0.414,P<0.01),与CD4、CD8+ T细胞表面CD38、CD95表达水平明显正相关(P<0.05),与CD4、CD8+ T细胞表面HLA-DR表达无显著相关性.结论 中国HIV感染LTNP CD4+CD25+Foxp3+ 调节性T细胞百分率明显低于典型进展者,提示调节性T细胞与LTNP保护机制相关.     

淮安市经异性性传播的HIV感染者中HIV-1病毒基因亚型特征分析

目的 了解淮安市经异性性传播HIV感染者中HIV-1病毒基因亚型分布及其流行特征.方法 从60名异性性传播HIV-1感染者血液中提取DNA或RNA,用巢式PCR或RT-PCR方法扩增gag、env基因区片段,测定序列并分析.结果 60份标本中,确定了48份标本的基因亚型,发现有4种HIV-1亚型和重组型.其中CRF01_AE亚型占62.5％,CRF07_BC亚型占22.9％,CRF08_BC亚型占6.3％,B亚型占6.3％.结论淮安市异性性传播感染HIV者中,流行着CRF01_AE、CRF07_BC、CRF08_BC、B亚型这4种亚型病毒株,CRF01_AE为主要流行亚型.     

中国HIV/AIDS患者T细胞及调节性T细胞CTLA-4表达与疾病进展关系研究

目的 对中国HIV感染者T细胞及凋节性T细胞CTLA-4表达与HIV疾病进展的相关性进行研究,探讨CTLA-4在HIV感染中的作用.方法 选取58名HIV/AIDS患者(长期不进展组、无症状HIV组、AIDS组),应用流式细胞仪胞内染色技术检测T细胞及CD4+CD25+Foxp3+调节性T细胞内CTLA-4表达水平,分析其与CD4+T细胞、病毒载量、淋巴细胞活化凋亡水平的相关性.结果 长期不进展组、无症状HIV组、AIDS组CD4+T细胞内CTLA-4表达水平依次增岛(P<0.05);与CD+T细胞显著负相关(P<0.01),与CD8+T细胞活化(CD38表达)、凋亡水平(CD95表达)及CD4+T细胞凋亡水平显著相关(P<0.05),与病毒载量无显著相关性.长期不进展组、无症状HIV组、AIDS组CD8+T细胞内CTLA-4表达水平差异无统计学意义;与CD4+T细胞、病毒载量、CD4,'+>、CD8+T细胞活化及凋亡水平均无显著相关性.CD4+CD25+Foxp3+T细胞内CTLA-4表达水平长期不进展组明显低于无症状HIV组及AIDS纽(P<0.05);与CD4+T细胞显著负相关(P<0.05);与CD4+、CD8+T淋巴细胞活化(HLA-DR表达)显著相关(P<0.01).结论 中国HIV感染者CD4+T细胞及CD4+CD25+Foxp3+调节性T细胞内CTLA-4表达水平与疾病进展及免疫活化状态显著相关,参与HIV感染免疫平衡的调节.     

HIV感染者外周血CD8~+NK细胞百分比及功能变化研究

目的探讨HIV感染者外周血中CD8~+NK细胞百分比的变化与表达CD107a的能力。方法选取未经高效抗逆转录病毒疗法(highly active antiretroviral therapy,HAART)治疗的HIV慢性感染者,感染者按照CD4~+T细胞数量分成高、低2组,用流式细胞仪检测其外周CD8~+NK细胞百分比的变化情况和CD107a的表达量。结果 HIV感染者外周血中CD8~+NK细胞的百分比明显低于正常人(P=0.011 1),CD8~+NK细胞的百分比与病毒载量成明显的负相关关系(r=-0.541 9,P=0.045 3)。K562细胞系刺激后,CD4~+T细胞高组CD107a的表达量明显增高。结论具有抗病毒作用的CD8~+NK细胞在HIV感染后百分比下降;CD8~+NK细胞水平高以及功能较强者HIV病毒水平相对降低,CD4~+T细胞数量相对高。     

中国经采供血HIV感染长期不进展者CD4~+T淋巴细胞趋化因子受体表达研究

目的:了解中国经采供血HIV感染长期不进展者CD4+T淋巴细胞趋化因子受体表达,分析其与疾病不进展的关系。方法:收集43例经采供血HIV感染长期不进展者、82例无症状HIV感染者、35例AIDS病人及40例健康对照的抗凝全血,用流式细胞仪检测趋化因子受体CCR5、CXCR4的表达,并分析其与病毒载量、CD4+T淋巴细胞绝对值及T淋巴细胞活化的相关性。结果:长期不进展组CD4+T细胞表面CCR5的表达明显低于无症状HIV感染组及AIDS组(P0.01),与健康对照无显著差异;CD4+T细胞表面CXCR4的表达各组无显著差异。CD4+T细胞表面CCR5的表达与CD4+T细胞数量显著负相关(r=-0.498,P0.05),与病毒载量无显著相关性。CD4+T细胞表面CCR5的表达与HLA、CD38在CD4+、CD8+T细胞的表达水平显著正相关(P0.001,CD38在CD4+T细胞的表达除外),CD4+T细胞表面CXCR4的表达与HLA在CD4、CD8+T细胞的表达水平显著负相关(P0.01)。结论:HIV感染长期不进展者CD4+T细胞趋化因子受体CCR5表达维持较低水平,与疾病不进展相关。     

细胞毒性T淋巴细胞增殖、分化能力对HIV/HCV重叠感染患者病情进展的影响

目的 探讨HIV/HCV重叠感染患者细胞毒性T淋巴细胞增殖能力和分化状态对病情进展的影响.方法 采用流式细胞仪检测CD57、CD27和CD28在CD8+T细胞上的表达,根据其表达差异,判断CD8+T细胞的增殖能力和不同的分化状态,比较HIV/HCV重叠感染者和单独HCV感染者增殖能力和分化状态的差异.探讨CTL功能与病情进展的关系.结果 HIV/HCV重叠感染者中CD57在CD8+T细胞上高表达28.84%±4.49%,而在HCV单独感染者中较低表达8.24%±5.05%,两组差异非常显著(P＜0.001).CD57+CD8+T细胞的百分数与HCV载量对数值间存在着正线性相关(P=0.023,R2=0.21).HIV/HCV重叠感染者的CTL细胞分化状态以晚期为主,而HCV单独感染者的CTL细胞分化状态以中期为主,两者比较差异非常显著(P＜0.001).结论 HIV/HCV重叠感染患者较HCV单独感染者CTL细胞增殖能力较低、呈终末期分化状态,进而影响CTL的免疫应答,可能是重叠感染者肝病进展的原因之一.     

北京市性传播HIV-1感染者流行毒株gag基因序列测定和亚型分析

目的 了解北京市性传播HIV-1感染者流行毒株亚型特点和流行规律.方法 随机采集北京市2008年新确证性传播HIV感染者的抗凝全血标本100份,分离血浆,提取病毒RNA,用套式聚合酶链反应扩增病毒gag基因,并进行序列测定和亚型分析.结果 系统进化分析确定北京市性传播HIV-1感染者流行毒株存在8个亚型或流行重组型,分别为B亚型22份,B'亚型8份,C亚型1份,CRF01_AE 38份,CRF02_AG 2份,CRF07 BC 9份,CRF08_BC 3份,疑似C/CRF01_AE重组型1份.结论 CRF01_AE和B亚型分别占45.2%和26.2%,为性传播感染者主要的亚型,应该加强我市HIV-1亚型流行情况的监测.

Abstract：

Objective To investigate the subtype distribution and sequence characteristics of HIV-1 strains prevalent among sexual infectors in Beijing. Methods We collected the blood samples from 100HIV sexual infectors in Beijing during 2008 and separated plasma specimens. RNA was extracted from the plasma and the gag gene was amplified by RT-PCR and nest-PCR. The PCR products were sequenced directly and phylogenetic analyses of gag gene was performed using the MEGA4 software. Results Among 100 HIV-1 plasma samples,84 gag gene fragments were amplified and analyzed. Eight HIV subtypes including B(22 strains), B'(8 strains),C( 1 strain) ,CRF01_AE (38 strains) ,CRF02_AG (2 strains) ,CRF07_BC(9 strains) ,CRF08_BC(3 strains) and C/CRF01_AE recombinant like strain( 1 strain) were identified circulating in Beijing. Conclusion CRF01 _AE and subtype B were predominant in Beijing account for 45.2% and 26.2% and the surveillance of HIV gene variation should be paid more attention.     

Mother to child transmission of HIV‐1 in a Thai population: Role of virus characteristics and maternal humoral immune response

The objective of this study was to investigate factors influencing mother to child transmission of HIV‐1 in Thailand, where HIV‐1 CRF01_AE, the major subtype in Southeast Asia, predominates. Samples from 84 HIV‐1 infected, anti‐retroviral treatment‐naïve, non‐breast feeding mothers, 28 who transmitted HIV‐1 to their babies (transmitters) and 56 who did not (non‐transmitters), were studied for maternal humoral immune response and virus characteristics. Maternal humoral immune response was measured by lymphocyte phenotyping; neutralizing antibodies to laboratory HIV‐1 MN strain and two clinical isolates; peptide binding antibody to gp41 and V3 from strains CRF01_AE, B, and MN; autologous antibodies; and quasispecies diversity. Virus characteristics studied were viral load, co‐receptor usage, and viral replication capacity. No significant difference between transmitters and non‐transmitters was found for any parameter of maternal humoral immune response. However, viral load and viral replication capacity were significantly higher in transmitters versus non‐transmitters and were not correlated with each other. This suggests that viral replication capacity may be a transmission factor independent of viral load, which is already well established as a risk factor for transmission of HIV‐1. All except four viral isolates used the CCR5 co‐receptor. This is one of few studies of vertical transmission in a population where HIV‐1 CRF01_AE predominates. The data suggest that in this population the maternal humoral immune response was not important in preventing transmission at parturition, but that virus characteristics were key factors, and that viral replication capacity may contribute to birth‐associated mother to child transmission of HIV‐1. J. Med. Virol. 81:768–778, 2009. © 2009 Wiley‐Liss, Inc.     

HIV coinfections with tuberculosis among HIV-1 infected individuals in old cross river state,Nigeria

ABSTRACT

Human immunodeficiency virus (HIV) and Tuberculosis (TB) are two main global public health threats that dent development in low and middle-income countries. This study evaluated the HIV/TB co-infection rate among HIV-1 infected individuals in old Cross River State, Nigeria. A total of 417 HIV-infected individuals participated in this study, 241 (57.8%) from Calabar, Cross River State, Nigeria and 176 (42.2%) from Uyo, Akwa-Ibom State, Nigeria. The age range of the 417 HIV-1 positive individuals who participated in the study was 4–72 years with an average age of 39.1 years. Plasma samples were analyzed for HIV and TB using fourth-generation Enzyme-Linked immunosorbent Assay. The CD4 count was enumerated using the Partec CyFlow® Counter. Plasma viral loads (PVL) were determined using the Abbott Real-Time HIV-1 assay. Results showed that 230 (55.2%) of the participants were in the 31–45 years age range. The majority (67.4%) of the HIV-1 infected individuals were females and 32.6% were males. An overall prevalence of HIV/TB coinfection in Old Cross River State, Nigeria was 1.4%, with Akwa Ibom State (0.6%) and Cross River State (1.2%). A higher prevalence of HIV/TB coinfection was observed among females (1.8%) than in males (0.7%). Higher prevalences of HIV/TB coinfections was observed in patients above 45 years of age (2.2%), married (2.3%), tertiary education (1.8%) followed by those with secondary education (1.4%), traders and civil servants (3.1%), patients with CD4 counts 200–349 and ≥500 cells/μl (1.9%), and those with viral load <40 copies/mL (2.7%). This study confirmed the presence of HIV/TB co-infection in old Cross River State, Nigeria. Although the prevalence rate of HIV/TB coinfection was low, its presence alone among HIV-1 infected individuals makes it a major source of concern. This finding highlights the need for a well-structured approach to the management of co-infection, and this includes both the social and medical aspects of the problem.     

Characterization of mutations in CRF01_AE virus isolates from antiretroviral treatment-naive and -experienced patients in Singapore

OBJECTIVE: To determine the frequency of drug resistance mutations among antiretroviral treatment-naive and -experienced patients infected with CRF01_AE virus. DESIGN: Prospective observational study. METHODS: We recruited antiretroviral-experienced HIV-infected patients with suspected drug resistance and consecutive newly diagnosed drug-naive patients. Viral sequencing was performed using standard methods. Frequencies of mutations in CRF01_AE virus isolates were compared with reference data for subtype B virus. RESULTS: Sequences were obtained for CRF01_AE virus isolates from 69 patients with treatment exposure and 35 treatment-naive patients. Treatment-naive patients had numerous polymorphisms but no major drug resistance mutations. There were differences between CRF01_AE and subtype B viruses in several drug resistance mutations including the following: D67N, L210F, K101E, V106M, V179I/D, G190A/S/E, and G48V (which were more common in CRF01_AE virus) and M41L, T215Y, and V82A (which were less common in CRF01_AE virus). CONCLUSIONS: The pattern of treatment-related mutations in CRF01_AE virus differs from that in subtype B virus at a number of positions determining drug resistance. Understanding these differences is important for interpreting results of resistance tests and determining treatment choices.     

2008年北京市HIV-1耐药株传播调查

目的 了解2008年北京市未经抗病毒治疗HIV-1感染者中耐药株传播水平,为耐药监测和临床抗病毒治疗工作提供本底资料.方法 参照WHO提出的HIV耐药警戒线调查方法(HIV drug resistance threshold survey,HIVDR-TS)指导方案,收集6个月内检测发现的60～70名小于25岁的感染者血浆样本,检测HIV-1 pol区亚型及耐药基因型,并计算耐药株检出率、评价传播水平.结果 61份符合要求的样本共获得50个有效pol区序列.感染途径以同性传播为主,占62%;亚型分布以B(42%)、CRF01_AE(28%)、CRF07_BC(26%)3种为主.出现1例针对PI类药物的耐药突变株,检出率为2%(1/50);出现1例针对NRTI类药物的耐药突变株,检出率为2%(1/50);未出现针对NNRTI类药物的耐药突变株,检出率为0.蛋白酶(PR)区和逆转录酶(RT)区的耐药突变株检出率均为2%,均属于低度传播范围(<5%).结论 北京市未经抗病毒治疗HIV-1感染者中出现PR和RT区的耐药突变株,传播水平尚处于低流行状态,现有的抗病毒治疗方案是可行的,治疗前尚不需要进行大规模耐药性检测.     

2008年北京市HIV-1耐药株传播调查

目的 了解2008年北京市未经抗病毒治疗HIV-1感染者中耐药株传播水平,为耐药监测和临床抗病毒治疗工作提供本底资料.方法 参照WHO提出的HIV耐药警戒线调查方法(HIV drug resistance threshold survey,HIVDR-TS)指导方案,收集6个月内检测发现的60～70名小于25岁的感染者血浆样本,检测HIV-1 pol区亚型及耐药基因型,并计算耐药株检出率、评价传播水平.结果 61份符合要求的样本共获得50个有效pol区序列.感染途径以同性传播为主,占62%;亚型分布以B(42%)、CRF01_AE(28%)、CRF07_BC(26%)3种为主.出现1例针对PI类药物的耐药突变株,检出率为2%(1/50);出现1例针对NRTI类药物的耐药突变株,检出率为2%(1/50);未出现针对NNRTI类药物的耐药突变株,检出率为0.蛋白酶(PR)区和逆转录酶(RT)区的耐药突变株检出率均为2%,均属于低度传播范围(<5%).结论 北京市未经抗病毒治疗HIV-1感染者中出现PR和RT区的耐药突变株,传播水平尚处于低流行状态,现有的抗病毒治疗方案是可行的,治疗前尚不需要进行大规模耐药性检测.     

2008年北京市HIV-1耐药株传播调查

目的 了解2008年北京市未经抗病毒治疗HIV-1感染者中耐药株传播水平,为耐药监测和临床抗病毒治疗工作提供本底资料.方法 参照WHO提出的HIV耐药警戒线调查方法(HIV drug resistance threshold survey,HIVDR-TS)指导方案,收集6个月内检测发现的60～70名小于25岁的感染者血浆样本,检测HIV-1 pol区亚型及耐药基因型,并计算耐药株检出率、评价传播水平.结果 61份符合要求的样本共获得50个有效pol区序列.感染途径以同性传播为主,占62%;亚型分布以B(42%)、CRF01_AE(28%)、CRF07_BC(26%)3种为主.出现1例针对PI类药物的耐药突变株,检出率为2%(1/50);出现1例针对NRTI类药物的耐药突变株,检出率为2%(1/50);未出现针对NNRTI类药物的耐药突变株,检出率为0.蛋白酶(PR)区和逆转录酶(RT)区的耐药突变株检出率均为2%,均属于低度传播范围(<5%).结论 北京市未经抗病毒治疗HIV-1感染者中出现PR和RT区的耐药突变株,传播水平尚处于低流行状态,现有的抗病毒治疗方案是可行的,治疗前尚不需要进行大规模耐药性检测.     

2008年北京市HIV-1耐药株传播调查

目的 了解2008年北京市未经抗病毒治疗HIV-1感染者中耐药株传播水平,为耐药监测和临床抗病毒治疗工作提供本底资料.方法 参照WHO提出的HIV耐药警戒线调查方法(HIV drug resistance threshold survey,HIVDR-TS)指导方案,收集6个月内检测发现的60～70名小于25岁的感染者血浆样本,检测HIV-1 pol区亚型及耐药基因型,并计算耐药株检出率、评价传播水平.结果 61份符合要求的样本共获得50个有效pol区序列.感染途径以同性传播为主,占62%;亚型分布以B(42%)、CRF01_AE(28%)、CRF07_BC(26%)3种为主.出现1例针对PI类药物的耐药突变株,检出率为2%(1/50);出现1例针对NRTI类药物的耐药突变株,检出率为2%(1/50);未出现针对NNRTI类药物的耐药突变株,检出率为0.蛋白酶(PR)区和逆转录酶(RT)区的耐药突变株检出率均为2%,均属于低度传播范围(<5%).结论 北京市未经抗病毒治疗HIV-1感染者中出现PR和RT区的耐药突变株,传播水平尚处于低流行状态,现有的抗病毒治疗方案是可行的,治疗前尚不需要进行大规模耐药性检测.