重组人β-趋化因子RANTES抗T嗜性HIV-1作用的研究

目的　检测重组人β 趋化因子RANTES(RegulationuponactivationnormalT cellexpressedandsecreted)对T嗜性HIV 1病毒株SF 33在T细胞中复制的影响。方法　在SF 33感染细胞前 1h ,用不同浓度 (5ng/ml、5 0ng/ml、5 0 0ng/ml)的重组人RANTES、GST RANTES或TEP RANTES对MT 4细胞进行预处理。病毒感染后第6天 ,以ELISA方法检测p2 4抗原水平 ,MTT比色法检测感染细胞的存活数。结果　在不同浓度下 ,天然RANTES及修饰RANTES对SF 33p2 4水平不产生显著影响 ,半数抑制浓度IC50 >5 0 0ng/ml,对感染细胞的保护率在 0以下。结论　T嗜性HIV 1病毒株SF 33对RANTES及其修饰蛋白的抑制作用不敏感。     

HIV-1 B'/C亚型毒株辅助受体的利用

目的分析疾病进展不同阶段分离的B’肥亚型毒株辅助受体的利用情况。方法从长期不进展者和AIDS期患者体内分离病毒株,将获得的B’／C毒株进行体外培养,用病毒株感染U87．CD4、GHOST细胞系并进行培养,通过检测、比较培养基上清液中核衣壳蛋白p24量的差异来确定某一毒株利用的辅助受体。再利用辅助受体抑制剂TAK～799、AMD-3100验证从细胞系实验中得到的结论。结果从AIDS期患者体内分离的毒株大部分利用C—C家族趋化因子受体（C—Cchemokinereceptor,CCR）5辅助受体,部分利用C—X～C家族趋化因子受体（C—X—C chemokine receptor,CXCR）4和CCR5辅助受体。从长期不进展者体内分离的毒株利用CCR5辅助受体。结论从长期不进展者体内分离的病毒主要利用CCR5辅助受体。从AIDS期患者体内分离的病毒主要利用CCR5辅助受体,也有同时利用CCR5和CXCR4辅助受体的现象。     

长期ART中HIV-1抗体的水平变化特点及其与HIV-1储存库相关关系的研究

目的 分析HIV-1感染者在长期抗反转录病毒治疗(antiretroviral therapy,ART)前后血浆中p24抗体和总HIV-1特异性抗体水平变化特点及其与HIV-1储存库关系.方法 选取2009年11月-2013年7月于我中心接受ART满5年的27例HIV-1感染者作为研究对象,采用荧光素酶免疫吸附试验和酶...     

早期诊断围产期HIV-1感染几种方法的比较

在一儿科AIDS临床试验里,早期诊断用齐多夫定治疗的婴儿HIV 1感染的三种方法是:1)血浆HIV 1RNA扩张试验(RNA试验) ;2 )外周血定量单核细胞(PBMC)的微量培养法(培养法) ;3)PBMCHIV 1DNA扩张试验(DNA试验)。这三种方法用于2 4名感染HIV 1的婴儿和10 0名非感染婴儿。HIV 1感染的定义是多于或等于2次培养阳性;在≥18个月时显示HIV 1抗体阳性。婴儿出生时和于6周与2 4周龄时进行PBMC培养;冷冻保存的标本则作RNA和DNA试验。婴儿出生时,培养方法、DNA和RNA试验的敏感性分别为2 0 8%、10 5 %和2 6 7%。年龄较大时,敏感性…     

HIV-1辅受体的配体在HeLa细胞的表达及其对合胞体形成的抑制

目的 观察HIV—1辅受体的配体、趋化因子RANTES和SDF—1的双顺反子表达载体pCMV—R—K—SK在HeLa细胞系表达，并对其抗HIV—1感染作用进行初步观察。方法 应用PCR扩增RANTES-KDEL基因，鉴定后与真核表达质粒pCMV—S／K连接，构建RANTES和SDF—1双顺反子表达载体pCMV—R—K—S—K，酶切鉴定并测序。脂质体介导转染HeLa细胞，间接免疫荧光及放射免疫沉淀法检测RANTES和SDF—1表达。合胞体形成实验初步检测其抗HIV—1感染的作用。结果 酶切鉴定和测序证明成功构建了pCMV—R—K—S—K双顺反子表达载体，间接免疫荧光及放射免疫沉淀法证实RANTES和SDF—I可以表达于HeLa细胞。pCMV—R—K—SK转染能够抑制M和T嗜性HIV—1膜蛋白诱导的合胞体形成。结论 双顺反子表达载体pCMV—R—K—S—K转染的HeLa细胞可以表达HIV—1辅受体的配体RANTES和SDF—1，并能抵抗HIV—1感染。     

HIV-1辅受体的配体细胞内双表达对病毒感染的阻断作用

目的　观察Ⅰ型人免疫缺陷病毒 (HIV 1)辅受体的配体———RANTES和SDF 1α双表达于人淋巴细胞对各种嗜性HIV 1毒株感染的阻断作用。方法　用 pLNCX R K S K重组逆转录病毒液感染原代人外周血淋巴细胞 (PBLs) ,抗神经生长因子受体 (NGFR) 免疫磁珠法分离转化成功的PBLs,流式细胞仪检测筛选效率 ;HIV 1M嗜性、T嗜性和双嗜性毒株攻击转化PBLs ,检测HIV 1逆转录酶活性和 p2 4抗原分泌 ,以观察抗HIV 1感染的作用 ;同时进行转化PBLs表面CD3、CD4、CCR2、CCR5和CXCR4表达及破伤风毒素刺激后3 H 胸苷 (thymidine)掺入量检测 ,观察HIV 1辅受体配体的双表达对人PBLs正常生物学功能的影响。结果　抗 NGFR 免疫磁珠法获得了转化成功的PBLs,流式细胞仪检测发现pLNCX R K S K转染组 92 %以上的PBLs鼠抗NGFR标记物为阳性 ;HIV 1M嗜性、T嗜性和双嗜性毒株攻击后 ,pLNCX R K S K转化PBLs可以见到明显的逆转录酶活性和 p2 4抗原分泌抑制 ,并且在感染后第 12～ 2 0天时抑制作用最强 ;pLNCX R K S K转化PBLs表面CD3、CD4和CCR2表达水平无明显变化 ,而CCR5和CXCR4表达水平降低 ;破伤风毒素刺激后的转化PBLs仍具有主动增殖的能力。结论　HIV 1辅受体的配体通过在人PBLs内双表达 ,使HIV 1两类主要辅受体表型剔除 ,基本阻断了     

不同病期HIV-1感染者的血浆中和抗体研究

目的了解HIV-1感染者体内中和抗体状况,探讨HIV-1感染的病理机制.方法病毒的分离和培养采用共培养(co-culture)方法,从29例HIV-1感染者的外周血淋巴细胞(PBMC)分离了29株HIV-1原代病毒,中和实验采用病例血浆中和分离的相应病毒株,再感染经PHA刺激活化后的正常人PBMC,培养5天检测其HIV-1 p24抗原含量以判断中和效果.结果29例研究时象中,24例是无症状感染者,5例为AIDS患者.我们发现17例无症状感染者的ID50≥8,其中10例ID90≥8,4例产生高滴度中和抗体ID90≥64,交叉中和实验结果表明这4例感染者的血浆对异体病毒均有不同程度的中和效应,其中尤以s7,s28作用最强.而AIDS患者中仅2例ID50≥8,其中1例ID90≥8,无1例产生高滴度中和抗体.结论通过对HIV-1感染者血浆中和效应的观察,表明多数感染者对自身同血源病毒的中和抗体滴度较低,但也有少数感染者能产生较高滴度的中和抗体,有必要深入研究其广泛的中和病毒效应;此外,早期无症状的感染者似比晚期有症状者对自体病毒的中和效应强,提示抗病毒治疗应于感染早期进行,以免机体体液免疫反应的进一步恶化.     

新喋呤水平预测晚期HIV-1感染的疾病进展

据医学空间网3月11日报道(原载Clin Infect Dis2005：40：853-858)，免疫激活的标志物-新喋呤血浆水平增高是晚期HIV-1感染病人疾病进展的可靠、独立预测因子。     

黑龙江省服刑人员HIV-1nef基因分析

目的研究黑龙江省服刑人员中HIV-1感染者病毒流行株的起源及其序列变异特征。方法采集黑龙江省16例感染HIV-1服刑人员的抗凝全血,分离血浆,用巢式聚合酶链反应对HIV-1附属基因(nef)进行扩增,并直接进行序列测定。应用BioEdit和MEGA软件进行序列及系统发育分析。结果系统进化树显示12例体内HIV的nef区序列属于泰国B亚型,3例属于重组型BC,1例属于重组亚型AE。各隔离株nef区的组内基因距离为13.53%。按传播途径分组,各组基因距离差异无统计学意义(P>0.05),表明nef区序列变异可能与传播途径无关。与国际标准株(HBX2)进行比较时,可发现某些nef序列的氨基酸变异。但是,大多数序列还是有较好的保守性。结论本组HIV-1病毒株可能有共同的起源,本文所讨论的氨基酸序列有望对nef功能区的未来研究提供参考信息。     

Kinetics of plasma cytokines and chemokines during primary HIV-1 infection and after analytical treatment interruption

Background

There are strong theoretical arguments for initiating antiretroviral therapy (ART) during primary HIV‐1 infection (PHI) to preserve HIV‐1‐specific T‐cell responses and to decrease immune activation.

Methods

We assessed the degree of immune activation during PHI and after analytical treatment interruption (ATI) in plasma samples from 22 subjects by measuring 13 cytokines/chemokines with the Luminex system. Subjects initiated quadruple ART at PHI (the QUEST cohort) and were classified as responders or nonresponders according to their HIV‐1 viral load (VL) 6 months post‐ATI.

Results

During PHI, nonresponders had higher levels of HIV‐1 RNA, interferon (IFN)‐γ, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐10 and eotaxin than responders (P≤0.05). A positive correlation was found between VL and IFN‐α, TNF‐α, IL‐1β, macrophage inflammatory protein (MIP)‐1α and MIP‐1β, respectively. Post ATI, responders had higher levels of IFN‐γ, MIP‐1β and monocyte chemotactic protein (MCP)‐1 than nonresponders, while nonresponders had higher levels of HIV‐1 RNA, IL‐15 and eotaxin. Cytokine/chemokine levels were higher during PHI than post‐ATI.

Conclusions

High levels of immune activation during PHI are associated with a worse virological outcome post‐ATI. In contrast, VL post‐ATI is negatively correlated with IFN‐γ and chemokines. Therefore, the degree of immune activation during PHI is associated with both the VL at PHI and the viral set‐point post‐ART.     

Interactions between schistosomiasis and infection with HIV-1

In many regions of the world, both schistosomiasis and HIV/AIDS are endemic, resulting in patients harbouring co-infections. Because interaction with host CD4(+) T cells is a characteristic of schistosome as well as HIV-1 infections, bi-directional disease effects may be sufficiently different from sequelae caused by either infectious agent alone to warrant alteration of public health approaches in areas of co-endemnicity. Studies published over the past decade provide useful insights into interactions between schistosomiasis and infection with HIV-1, and overall support the hypothesis that special emphasis on treatment of schistosomiasis in populations with elevated prevalence or risk of HIV-1 infection is justified.     

HIV-1慢性感染者B细胞功能受损的临床意义

HIV慢性感染是全球性的问题和大家关注的热点,其主要标志是CD4+ T淋巴细胞计数的减少和免疫系统的功能失调.关于其发病机制方面的研究很多,但是关于B细胞方面的研究较少,而且关于其在HIV慢性感染中的功能和意义尚不清楚.本文对HIV-1慢性感染者B细胞功能受损的临床意义进行综述,以期为HIV感染的研究工作提供参考.     

Neuropsychological performance in patients with asymptomatic HIV-1 infection

Human immunodeficiency virus (HIV-1) infection and acquired immunodeficiency syndrome (AIDS) lead to neurocognitive disorders; however, there is still much knowledge to be gained regarding HIV-associated neurocognitive disorders. The purpose of this study was to assess the cognitive performance, instrumental activities of daily living, depression, and anxiety in patients with asymptomatic HIV-1 infections compared with seronegative participants without neurocognitive impairment. We studied a sample consisted of 60 patients with asymptomatic HIV-1 infections and 60 seronegative participants without neurocognitive impairment from the city of Barranquilla, Colombia, with a mean age of 36.07 years. A protocol of neuropsychological and psychopathological tests was applied to the participants. The group of patients with asymptomatic HIV infections significantly underperformed on tasks that assessed global cognitive screening, attention span, learning, phonemic verbal fluency, auditory-verbal comprehension, information processing speed, cognitive flexibility, and motor skills compared to the group of seronegative participants. No significant differences were found in memory, visual confrontation naming, vocabulary, inhibition, and instrumental activities of daily living. Additionally, the patients with asymptomatic HIV-1 infection had a higher anxiety index than the seronegative participants, but no significant difference was found in depression. A correlation was found between depression and anxiety. In conclusion, the patients with asymptomatic HIV-1 infection had lower cognitive performances than the seronegative participants in the cognitive functions mentioned above and more anxiety but still performed the instrumental activities of daily living.     

HIV-2 antibody testing of blood donors with doubtful immunoblot results for HIV-1

Summary Antibodies against human immunodeficiency virus type-1 (HIV-1) in samples from blood donors are commonly detected by various enzyme-linked immunosorbent assays (ELISA) and by confirmatory tests, e.g., Western blot or immunofluorescence tests. Immunoblot reactivity, which is directed only towards the HIV-1 core proteins p 18, p 24 and p 55, may represent false-positive reactions. Out of 125,000 blood donations, 140 were repeatably HIV-1 antibody reactive by ELISA; of these, 20 were doubtful positive sera with isolated p 18 and/or p24 bands in the HIV-1 confirmatory assay. Antibodies to HIV-2 are known to cross-react with these HIV-1 core proteins. We therefore assayed the 20 sera by immunofluorescence and immunoblotting for the presence of antibodies to HIV-2. None of these doubtful HIV-1 antibody positive blood donor sera was found to have antibodies to HIV-2.     

PNS外周血单核细胞趋化因子与RANTES mRNA的表达及意义

目的 探讨原发性肾病综合征(PNS)患者外周血单核细胞趋化因子(MCP-1)与受激活调节的正常T细胞表达和分泌因子(RANTES)mRNA的表达及临床价值。方法 PNS患者54例，分别于治疗前及治疗后3个月采用逆转录聚合酶链反应(RT—PCR)方法检测外周血MCP-1及RANTES mRNA的表达水平。结果 PNS患者外周血MCP-1、RANTES mRNA的表达显著高于正常对照组。治疗后显著降低；其中膜性肾病患者表达水平较低，系膜毛细血管性肾炎患者表达水平较高；伴有肾小管间质损害者表达水平显著高于无肾小管间质病变者，治疗后未完全缓解者显著高于完全缓解署。MCP-1、RANTES mRNA表达水平与血肌酐水平呈显著正相关，与24小时尿蛋白定量、血清白蛋白水平无显著相关性。结论 PNS患者外周血MCP-1、RANTES mRNA表达水平与病理类型、肾小管间质损害、血清肌酐水平及疗效有一定关联，对其检测有助于PNS病情及预后的评估。     

HBV感染对HIV-1病人抗病毒药物治疗影响的初步分析

目的了解合并乙型肝炎病毒(HBV)感染及其基因亚型对艾滋病病毒1型(HIV-1)病人抗病毒药物治疗疗效的影响。方法采用实时荧光定量聚合酶链反应(PCR)检测HBV基因型,根据分型情况进行组内比较。调查病人纳入治疗组时和进行抗病毒治疗(ART)6个月、12个月、24个月以及36个月后各项临床指标。结果共收集46份HIV-1、HBV合并感染病人的标本,以50份单独感染HIV-1病人的标本作为对照。46例共感染病人中,共检出30例B基因型,其余为非B基因型。每组病人经过治疗后,CD4细胞计数值均明显上升,肝脏生化酶指标均明显下降。比较B型与非B型病人的肝、肾功能指标,在治疗前期差异有统计学意义;但其他病毒学应答在ART治疗6个月、12个月、24个月以及36个月时,差异无统计学意义,比较单独感染HIV-1和HIV-1/HBV合并感染病人在治疗过程中的血小板变化,治疗6个月时的血肌酐,以及治疗12个月时的血红蛋白值,差异有统计学意义,其他病毒学应答无统计学意义。结论 ART治疗可明显提升共感染病人的免疫水平,促进丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)恢复正常,使病人病情好转。是否感染HBV和所感染HBV的基因型,在短期内可能影响到HIV-1病人的部分临床应答,但这种影响会随着治疗的深入而逐渐消失。     

应用流式细胞术检测HIV-1感染者外周血p24、gp41和gp120抗原表达的研究

目的分析HIV-1感染者外周血CD4^＋T淋巴细胞p24、gp41和gp120抗原的表达状况,探讨流式细胞术用于检测HIV-1感染的可行性。方法应用三色流式细胞术,对192例HIV-1感染者和29例健康者外周血CD4^＋T淋巴细胞表达的HIV-1 p24、gp41和gp120抗原进行检测。结果HIV-1 p24、gp41和gp120抗原阳性率HIV-1感染组与健康对照比较差异有统计学意义（P均〈0.01）;CD4^＋T细胞数〈200 cells/μl的HIV-1感染人群与CD4＋T细胞数〉200 cells/μl的人群比较差异有统计学意义（P〈0.01或P〈0.05）;静注吸毒途径感染人群与性途径感染人群比较差异有统计学意义（P〈0.01）。在HIV-1感染人群中,p24、gp41和gp120抗原阳性率四分位数25%75%区间范围分别为1.67%5.95%、1.61%8.12%和0.56%2.35%。结论流式细胞术简便、快速、敏感,可用于HIV-1感染检测。     

Hepatitis C virus infection among HIV-1 infected individuals from northern Mexico

Aims: The prevalence of hepatitis C virus (HCV) infection, risk factors and HCV genotypes in 140 HIV-1 infected individuals from northern Mexico was determined. Methods: Hepatitis C infection was confirmed by the detection of anti-HCV antibodies and HCV-RNA in sera, and genotyping was performed by the InnoLiPA-HCV genotype assay. Results: Seventeen (12.1%) out of 140 HIV-infected individuals were found to be HCV-positive. Coinfected individuals were more likely to be male (87%). The most frequent genotype was 1a (41%), followed by 1b (29.4%), 2a/c (17.6%), 2b (5.9%) and 3 (5.9%). Serum transaminase concentrations (AST and ALT) were higher in coinfected patients. Among the risk factors for coinfection: sexual transmission was the most frequently observed (men who have sex with men (MSM); 64.7% and bisexual behavior; 64.7%) followed by intravenous drug users (IVDU) (53%). There was no association of the HCV genotypes with the age and risk factors for HIV-1 and HCV infection observed in the studied patients. Conclusion: The results suggest that the prevalence of HIV-1/HCV coinfection in Mexico is lower than in other American countries.