双抗原夹心法检测抗HIV-1/2总抗体方法的建立和评价

目的 进一步提高HIv感染诊断试剂的敏感性。方法 以人工合成的HIv—1 gp41．1(sP1)、gp41．2(sP2)、gp120(sP3)、P24(sP4)和HIV—2gp36(sP5)5条多肽，采用双抗原夹心酶链免疫吸附试验(ELISA)原理，以sP1、sP3、sP4和sP5混合包被酶标板作为因相抗原，辣根过氧化物酶标记sp1、sp2、sP4和sP5多肽为标记物，建立了检测抗HIV—1／2总抗体的双抗原夹心ELISA法。结果 检测卫生部药品生物制品检定所第2代40份质控参比血清，其特异性和灵敏度均为100％，高于间接ELISA法(特异性为90％，灵敏度为65％)。检测210份其他病种患者血清均为阴性，与雅培HIVAB试剂比较检测如份健康献血员血清和88份HIv感染者血清，符合率为100％。结论 本方法特异性强、敏感性高，操作简便，适用于献血员的筛选和临床HIv感染的检测。     

辅受体突变导致HIV感染抑制的机理研究

目的 研究抗HIV感染者淋巴细胞HIV－1辅受体突变的情况。阐明其感染抑制的分子机理。方法 设计特定的引物，用PCR方法检测健康人，HIV感染者和抗HIV感染者的基因组DNA中HIV辅受体（CCR5）是否突变。结果 抗HIV感染者发生CCR5△32纯合子突变，健康人及HIV感染者CCR5基因未发生突变。讨论发现了我国本土人群存在CCR5△32纯合子突变，此突变可能为抗HIV感染保护机制，为艾滋病的治疗及预防提供新的方法和途径。     

某艾滋病治疗示范区人免疫缺陷病毒感染者合并隐匿性乙型肝炎病毒感染的调查分析

目的 调查分析某艾滋病治疗示范区人免疫缺陷病毒（HIV）-1感染者中隐匿性乙型肝炎病毒（HBV）感染的情况及其影响因素.方法 采集某艾滋病治疗示范区97例经血感染HIV-1的感染者的血浆,采用酶联免疫吸附试验（ELISA）检测乙型肝炎表面抗原与抗体（HBsAg与抗HBs）、乙型肝炎e抗原与抗体（HBeAg与抗Hbe）、乙型肝炎核心抗体（抗HBc）及丙型肝炎抗体（抗HCV）;采用吸附柱法抽提HBV DNA;采用巢式聚合酶链反应（PCR）法检测HBV S区;采用流式细胞仪计数CD4＋T淋巴细胞.HBsAg阴性PCR阳性结果 者为合并隐匿性HBV感染者.合并隐匿性HBV感染者为实验组,未合并隐匿性HBV感染者为对照组.结果 97例HIV感染者中HBsAg阴性者92例（94.85%）.92例HBsAg阴性者中合并隐匿性HBV感染者27例（29.35%）,抗HCV阳性者73例（79.35%）.合并隐匿性HBV感染者和未合并HBV感染者CD4＋T淋巴细胞数、单独抗HBc阳性率分别为（212.11±133.1）和（318.9±172.2）cells/mm3、62.96%和18.46%,以上两指标两组比较差异均有统计学意义（P＜0.01）,两组间年龄、性别、是否合并HCV感染及抗HBs阳性率比较差异无统计学意义（P＞0.05）.结论 经有偿献血途径感染HIV者中存在隐匿性HBV感染;HIV阳性合并隐匿性HBV感染者中易出现单纯抗HBc阳性;CD4＋T淋巴细胞数低的HIV感染者更容易合并隐匿性HBV感染.     

人免疫缺陷病毒感染者/艾滋病患者浅表淋巴结肿大与CD4+T淋巴细胞计数的相关性

目的 探讨人免疫缺陷病毒感染者/艾滋病患者（简称HIV感染者/AIDS患者）浅表淋巴结肿大的病理改变及其与CD4+T淋巴细胞计数的相关性。方法 对1066例HIV感染者/AIDS患者浅表淋巴结肿大的发生情况及其外周血CD4+T淋巴细胞计数进行分析;并对浅表淋巴结肿大患者行淋巴结活检。结果 在1066例HIV感染者/AI...     

Th22细胞及其关键转录因子在HIV/AIDS患者外周血中的表达及与病程进展的关系

目的分析HIV感染者或AIDS患者(HIV/AIDS)外周血单个核细胞(PBMC)中Th22细胞的含量及其关键转录因子芳香烃受体(Ah R)的表达量,探讨Th22细胞在HIV/AIDS疾病发生发展中的作用。方法筛选2014年3月至10月期间河南中医学院第一附属医院艾滋病研究中心经HIV抗体确证实验阳性者60例,根据CD4+~T细胞数将HIV/AIDS患者分为A(CD4200 cell/μl)、B(200≤CD4≤500 cell/μl)、C(CD4500 cell/μl)3组,同时选取体检中心健康人20例作为健康对照组,采用流式细胞术(FCM)检测外周血CD4+~T细胞数以及Th22细胞(CD4+~IL22+~)的百分比。RT-PCR技术检测其外周血单个核细胞(PBMC)中转录因子Ah R m RNA的相对表达量。结果 A组Th22细胞比率大于其他3组(P0.05);B组和C组Th22细胞比率均高于健康对照组,差异有统计学意义(P0.05);B组和C组Th22细胞比较无统计学意义(P0.05);A、B、C 3组转录因子Ah R m RNA相对表达量均明显高于健康对照组,差异有统计学意义(P0.05)。结论 HIV/AIDS患者PBMC中高表达Th22细胞及其转录因子Ah R m RNA,提示Th22细胞很可能与HIV/AIDS疾病发生发展关系密切。     

HIV感染者及其家属的心理状况调查研究

目的调查山西省某县艾滋病比较集中的村落的HIV感染者及其家属的心理状况，为艾滋病的干预和决策提供相关参考。方法采用焦虑自评量表（SAS）、抑郁自评量表（SDS）、生活质量综合评定量表（GQOLI）以及社会支持评定量表（SSAS）．对H1V感染者、HIV感染者的家属、未感染者（对照组）进行调查。结果①经血传播HIV感染者及其家属的主观最大压力来源是经济问题；②在焦虑、抑郁、生活质量以及支持总分上，感染组、家属组与对照组之间的差异都具有统计学意义。结论①提高当地人群的知识文化水平、发展当地经济会在一定程度上减少艾滋病感染及相关心理问题的发生；②经血传播HIV感染者及其家属的心理状况较差，有必要对他们实施心理干预。     

基于CiteSpace HIV感染者生育意愿研究可视化分析

目的 分析HIV感染者生育意愿研究热点和趋势，以期为HIV感染者生育相关研究提供参考。方法 在Web of Science核心合集数据库和中国知网（CNKI）中检索相关研究。运用CiteSpace 6.2.R4软件对发文量、作者、发文机构及关键词进行可视化分析。结果 本研究共纳入236篇文献。年发文量趋势平稳，高峰期出现在2014年，研究作者合作网络规模较大，研究机构主要以高校为主，各高校合作较紧密；研究热点集中于HIV传播、女性人群、干预治疗等方面。结论 抗逆转录病毒治疗是HIV感染者生育意愿研究的未来趋势。我国相关研究较少，国内学者和研究机构应借鉴国际的成熟经验，加强我国HIV感染者管理并发展HIV感染者生育意愿的相关研究。     

可溶性白细胞介素Ⅱ受体在爱滋病研究中的意义

可溶性白细胞介素-2受体(SIL-2R)在艾滋病研究中具有重要意义。血清IL-2R 水平升高是HIV 感染的特征之一,其升高程度与HIV 感染后的不同病期呈正相关,是动态监测HIV 感染后疾病进展情况的一项有价值的临床参数。HIV 感染者SIL-2R 水平与T 细胞功能呈负相关,不仅是HIV 感染者细胞功能受损的早期特征之一,其水平还反映了T细胞功能障碍的严重程度。此外,IL-2R 在一定程度上还可作为一项评价药物能否调节或增强AIDS 患者细胞免疫功能的指标。本文还就HIV 感染者SIL-2R 升高的可能机制作了初步探讨。     

脑脊液和血清HIV病毒载量的相关性研究

目的：评价HIV患者艾滋病期脑脊液与外周血病毒载量的相关性。探讨HIV感染后不同疾病阶段患者的脑脊液病理改变。方法选取HIV感染者116例。根据艾滋病患者血清中病毒载量,将研究对象按血清病毒载量分为HIV高、中、低病毒载量三组,以方差分析进行统计。结果随着疾病的进展,HIV感染后进入艾滋病期后,高病毒载量患者脑脊液HIV病毒载量与低、中组均存在差异。结论 HIV感染后,患者脑脊液中出现病理改变并与血清病毒载量的高低有关,脑脊液中HIV病毒载量即是这种改变的具体体现。     

HIV感染症状长期不进展者NK细胞变化研究

目的探讨HIV长期不进展者NK细胞的变化. 方法应用流式细胞术对HIV长期不进展者、典型进展者和HIV-抗体阴性正常对照外周血NK细胞、NKT细胞及NK细胞趋化因子受体等进行研究. 结果长期不进展者NKT细胞绝对计数与正常对照差异无统计学意义(P＝0.301),高于HIV感染者和艾滋病病人(P＝0.01, P＝0.002);长期不进展者NK细胞绝对计数低于正常对照(P＝0.03),高于HIV感染者和艾滋病病人(P＝0.005, P＜0.0001);长期不进展者NK细胞与CD4+ T淋巴细胞呈正相关(r＝0.393,P＝0.001);NKT细胞与CD8+ T淋巴细胞呈正相关(r＝0.372,P＝0.002).长期不进展者NK细胞表达的CCR5受体低于典型进展者和正常对照(P＜0.01). 结论 NK细胞的变化与HIV疾病进展相关,值得深入研究.     

Development and studies of the anti-R7V neutralizing antibody ELISA test: a new serological test for HIV seropositive patients

A new peptide-based ELISA test developed for the detection of anti-R7V specific antibodies in the sera of HIV seropositive patients is described. HIV virus acquires a cellular antigen at the moment the virus is released by budding, the R7V epitope. Certain patients produce anti-R7V Ab which are described as having the capacity to neutralize in vitro cell infection by HIV. Anti-R7V Ab are also detected in asymptomatic patients who have a lower likelihood of progressing to AIDS. Tested with 449 serum samples, the prevalence of anti-R7V Ab was 53.2% in HIV positive patients and 5.5% in HIV negative subjects. According to the duration of infection, the seroprevalence reaches almost 80% of non-treated long-term infected patients. Other retrospective studies were conducted on 308 HIV positive samples; the presence of anti-R7V Ab was significantly higher for 177 asymptomatic patients (64.4%) compared to the 131 symptomatic patients (35.1%). Regarding their neutralizing ability, anti-R7V antibodies were detected at the highest percentage in asymptomatic HIV-infected patients naive of treatment. Besides conventional biological parameters (CD4 and viral load), the detection of anti-R7V antibodies could be proposed to clinicians as an additional tool to manage treatment initiation and to improve the psychological state of their asymptomatic patients.     

HIV/AIDS感染者体外诱导的γδT细胞亚群的特点

目的:了解HIV/AIDS感染者外周血中γδT细胞亚群的特点以及体外诱导后γδT细胞的增殖情况和Vδ1、Vδ2亚群的变化,为γδT细胞体外扩增方法的建立打下基础。方法:流式细胞术(FCM)检测25例HIV/AIDS感染者(HIV组)和31例健康对照者(HC组)外周血中的γδT细胞、Vδ1亚群、Vδ2亚群的频率和绝对值;选择两组中各10例外周血单个核细胞(PBMC),用anti-γδTCR单克隆抗体(mAb)和IL-2在体外诱导培养14 d,在0 d、7 d、14 d分别进行细胞计数和FCM检测,分析比较γδT细胞亚群的比例和培养情况。结果:HIV组外周血中γδT细胞、Vδ2亚群百分比和绝对值都显著低于HC组,而Vδ1亚群百分比和绝对值显著高于HC组;体外诱导培养14 d时,HC组总细胞数量增加3倍,HIV组总细胞数量稍有增加;HC组的γδT细胞比例可达到80%以上,其中Vδ2亚群可以增加到65%左右,而HIV组的γδT细胞比例达35%左右,Vδ2亚群比例仅增加到17%左右。结论:HIV/AIDS感染者外周血中γδT细胞比例显著降低,Vδ1/Vδ2比值倒置;用anti-γδTCR mAb和IL-2在体外诱导HIV/AIDS感染者的Vδ2亚群扩增效果不佳,不能逆转Vδ1/Vδ2倒置现象。应进一步寻找更加有效的诱导培养方法。     

HLA多态性与HIV感染及AIDS发病相关性的研究

为研究人类白细胞抗原（ＨＬＡ）和人免疫缺陷病毒（ＨＩＶ）易感性的关系，本文分析比较了以下三组人群的ＨＬＡ表型频率和单倍型频率：①１７２例正常人；②１７例血清ＨＩＶ阴性高危人群；③１８０例血清ＨＩＶ阳性患者，其中２１例发展为艾滋病（ＡＩＤＳ），３７例６个月内ＣＤ４＋淋巴细胞降低至少２０％（ＣＤ４ｄｅｃｌｉｎｅ）。在１７２例对照和１８０例血清ＨＩＶ阳性受试者的比较中发现，其ＨＬＡ表型频率和单倍型频率没有显著差别，提示ＨＩＶ感染与ＨＬＡ无关联。然而，ＨＬＡ－Ｂ２１、ＨＬＡ－Ｂ８、ＨＬＡ－Ｂ３５表型及ＨＬＡ－Ａ１－Ｂ８、ＨＬＡ－Ｂ８－ＤＲ３、ＨＬＡ－Ａ１－Ｂ８－ＤＲ３单倍型与ＣＤ４阳性淋巴细胞下降，或与血清ＨＩＶ感染发展成ＡＩＤＳ病显著相关。１７例ＨＩＶ血清阴性高危人群中，无一携带单倍型ＨＬＡ－Ａ１－Ｂ８－ＤＲ３，提示该单倍型可能与ＨＩＶ感染的抗性相关。     

A response to P.H. Duesberg with reference to an idiotypic network model of AIDS immunopathogenesis

Professors in British Columbia, Canada have devised an autoimmunity model of AIDS (MIAMI model) based on an atypical network theory of regulation of the immune system. It presents different stimuli as cofactors for AIDS: allogenic stimuli in some risk groups and MHC mimicking antigenic stimuli in other risk groups. V regions are on helper T cells that are somewhat anti-self class II MCH. Helper T cell idiotypes interact with both class II MHC and particular suppressor T cell idiotypes, therefore both may be similar. In fact, foreign lymphocytes also can induce an immune response similar to that of MHC image (MI). Hence the MI response is against the anti-self MHC, i.e., foreign idiotypes identify self, particularly self MHC. Further, the HIV envelope protein (gp120) binds to CD4 at a site that overlaps the site where CD4 interacts with class II MHC. Thus recombinant gp120 well as antibodies that recognize the gp120 undying site of CD4 can prevent the interaction of CD4 with class II MHC. Some mutations of CD4 effect the gp120 binding site but not the class II binding site and vice versa and others effect both. This similarity and others HIV can be considered an image of class II MHC, and the anti HIV immune response may be anti MHC image (AMI). MI and AMI responses are against each other and against idiotypic determinants expressed on helper and suppressor T cells respectively. A dual attack on the idiotypes of helper and suppressor T cells accompany these responses thereby causing an imminent collapse of the entire immune system. The model's significant predictive power thereby suggests that we may be able to prevent HIV from causing AIDS by inducing immunological tolerance to HIV components that resemble MHC molecules. This model rejects the 11 paradoxes identified by Duesberg who surmises that HIV is not a cofactor or cause of AIDS.     

单核细胞TLR7/8表达与HIV-1感染疾病进程关系的研究

目的 对HIV-1感染者单核细胞TLR7/8表达水平及与疾病进展的相关性进行研究,探讨单核细胞TLR7/8在HIV-1疾病进程中的作用.方法 选取63名HIV-1感染者及18名健康对照,应用MACS磁珠分选法纯化CD14+单核细胞,用2.5μg/ml R848刺激单核细胞TLR7/8,QIAGEN公司的RNA提取试剂盒提取单核细胞总RNA,实时定量RT-PCR测定TLR7/8的mRNA表达.结果 HIV-1感染者单核细胞TLR7和TLR8的表达水平与CIM+T细胞显著正相关(r=0.614,P＜0.01;r=0.419,P＜0.01).TLR7 mRNA表达:缓慢进展组明显高于HIV感染组、AIDS组和健康对照组(P＜0.05),HIV感染组明显高于AIDS组(P＜0.05);TLR8 mRNA表达:缓慢进展组明显高于AIDS组(P＜0.05).体外用R848刺激11LR7后表达显著下调,而TLR8的表达稳定.结论 AIDS疾病进程中HIV-1感染者单核细胞TLR7/8表达水平明显下降,TLR7的表达水平下降更显著.     

HIV-1准种的变化与疾病进展关系的研究

目的　研究HIV 1准种的变化对疾病进展的影响。方法　PCR方法扩增HIV 1V3～V5区片段 ,采用HMA(异源双链泳动分析 )的方法分析 4 8例HIV/AIDS准种的变化 ,并结合HIV/AIDSCD4 T淋巴细胞计数及病毒载量的结果分析HIV 1准种的变化对疾病进程的影响。结果　在不同的疾病状态下 ,体内病毒变异是不相同的 ,在体内CD4 T淋巴细胞计数较高 ,血浆病毒载量低的个体内 ,病毒毒株有很强的变异特征 ,体内具有很高的准种复杂度 ;而在CD4 细胞数很低 ,病毒载量较高的个体 ,病毒基因变异较少 ,病毒准种较为单一。结论　HIV 1准种的变化与疾病进展相关。     

Human immunodeficiency virus (HIV) infection in haemophiliacs: long-term prognostic significance of the HIV serologic pattern.

To identify markers of prognostic value in the course of HIV disease, immunologic parameters and profiles of HIV antibodies and antigen were studied in 60 haemophiliacs. The 43 HIV-seropositive subjects were followed prospectively over a 4 year period with a retrospective analysis as well of their frozen plasma for HIV markers. This group had a significant decrease in number of helper/inducer T lymphocytes as compared with 17 HIV seronegative subjects. The degree of changes correlated with the stage of disease, with the most severe depletion of CD4 cells in those who developed AIDS. Counts of B cells and platelets were also lower in HIV-infected haemophiliacs. Ten out of 12 AIDS patients had undetectable antibodies to HIV p24 antigen; low levels of p24 antibody were also seen in six out of 15 subjects with lymphadenopathy (CDC stage III), but in only two out of 16 asymptomatic subjects (CDC stage II). Sustained HIV p24 antigenaemia (greater than 30 pg/ml) was seen in 10 AIDS patients, in five subjects with lymphadenopathy and in two asymptomatic haemophiliacs. Initial HIV serologic profiles, obtained when all patients were asymptomatic, were highly predictive for progression of the HIV infection: the initial pattern of low anti-p24 antibody and positive p24 antigenaemia conferred the worst prognosis, with all patients in this group developing ARC or AIDS within 36 months, whereas an initial high level of anti-p24 without p24 antigenaemia was associated with relatively the best prognosis. Of such subjects, 58% have remained clinically asymptomatic after 48 months of the study (P less than 0.00001). The serologic profile of HIV antibody pattern and HIV antigen in haemophilic patients thus already provides important prognostic information at an early stage of HIV infection.     

Inhibition of HIV replication by a CD4‐reactive Fab of an IgM clone isolated from a healthy HIV‐seronegative individual

HIV replication is restricted by some anti‐CD4 mouse mAb in vitro and in vivo. However, a human monoclonal anti‐CD4 Ab has not been isolated. We screened EBV‐transformed peripheral B cells from 12 adult donors for CD4‐reactive Ab production followed by functional reconstitution of Fab genes. Three independent IgM Fab clones reactive specifically to CD4 were isolated from a healthy HIV‐seronegative adult (～0.0013% of the peripheral B cells). The germ line combinations for the V_H and V_L genes were V_H3‐33/_L6, V_H3‐33/_L12, and V_H4‐4/_L12, respectively, accompanied by somatic hypermutations. Genetic analysis revealed a preference for V‐gene usage to develop CD4‐reactive Ab. Notably, one of the CD4‐reactive clones, HO538‐213, with an 1×10^?8 M dissociation constant (Kd) to recombinant human CD4, limited the replication of R5‐tropic and X4‐tropic HIV‐1 strains at 1–2.5 μg/mL in primary mononuclear cells. This is the first clonal genetic analysis of human monoclonal CD4‐reactive Ab. A mAb against CD4 isolated from a healthy individual could be useful in the intervention of HIV/AIDS.     

Free and antibody-complexed antigen and antibody profile in apparently healthy HIV seropositive individuals and in AIDS patients

The pattern of free and antibody-complexed HIV antigen and the antibody profile were investigated retrospectively in 305 serum samples taken from 22 AIDS patients before and during the development of AIDS and from 40 apparently healthy seropositive individuals. Most AIDS patients were found positive for both free and complexed antigen and had high gp41 antibody titres but low or undetectable p24 antibody. Four different patterns of HIV antigenaemia were observed: 1) positive for both free and complexed antigen; 2) negative for free HIV antigen at first, but always positive for complexed antigen; 3) positive for free antigen without complexed antigen; and 4) negative for both free and complexed antigen. The development of immune complexes preceded the appearance of free antigen and might reflect the ongoing viral replication with antigen excess and binding of anticore antibodies. No correlation was found between the development of AIDS symptoms and either the duration of free antigen positivity or the level of antigenaemia. A different pattern was observed in apparently healthy seropositive individuals: 90% of whom had high antibody titres to p24 and gp41 and were persistently negative for free and complexed HIV antigen. This study demonstrates that testing HIV markers in sequentially collected serum samples from HIV seropositive individuals is a useful and simple tool for early identification of persons at risk of developing AIDS.     

Activation and Coreceptor Expression of T Lymphocytes in HIV/AIDS Patients of China

The objective of this paper was to investigate the activation and coreceptor CCR5, CXCR4 expression of T lymphocytes in HIV/AIDS patients of China, and to study their association with disease progression. Seventy-seven HIV/AIDS patients and thirteen normal controls were enrolled and three-color flow-cytometry was used to detect the activation marker HLA-DR, CD38 and the coreceptor CCR5, CXCR4 expression on T lymphocytes in whole blood samples taken from the patients and the controls. The HLA-DR, CD38 and CCR5 expression on CD4, CD8⁺ T cells in AIDS patients was higher than in asymptomatic HIV-1 infected patients and normal controls (p < 0.05); The activation and CCR5 expression on T lymphocytes significantly correlated with CD4⁺ T lymphocyte number and viral load. The activation on T lymphocytes and the expression of CCR5 on T lymphocytes in HIV/AIDS patients of China are significantly correlated with disease progression.