HIV-1潜伏感染及功能性治愈

尽管高效抗反转录病毒治疗(HAART)可有效控制艾滋病(AIDS)病人体内的艾滋病病毒1型(HIV-1)的复制,但却无法根除潜伏感染的病毒,这成为当前艾滋病治疗的主要难点之一。研究HIV-1在宿主细胞内建立和维持潜伏的分子细胞学机制,有助于发现新的抗病毒靶点和发展新的抗病毒治疗策略。近年来对HIV感染者/AIDS病人提出功能性治愈策略,相关的免疫或基因治疗手段被相继提出,部分策略已处于临床试验阶段。该文对HIV-1潜伏感染机制和功能性治愈相关研究进展进行简要综述。     

MicroRNAs对HIV-1复制的调节作用及应用展望

微小核糖核酸(miRNAs)一种非编码小RNAs,在1型艾滋病病毒(HIV-1)感染的发展和转归中发挥重要作用。miRNAs通过直接作用于HIV RNA,或者通过间接调节细胞辅助因子和抗病毒因子抑制病毒复制,miRNAs也可以通过调节T细胞活化促进HIV感染。本文主要回顾在HIV-1感染者体内,异常表达的miRNAs通过多种途径促进病毒感染或者维持潜伏感染,及在感染过程中的生物调节作用。进一步研究miRNAs及其作用靶点在HIV-1感染者体内的作用可能为抗病毒治疗开启一个新的窗口。     

HIV早期感染检测及其使用策略

在艾滋病病毒(HIV)早期感染阶段,感染者体内会出现高病毒血症和高抗原血症,具有较强的传染性,但不能通过抗体检测做出诊断。早期感染的检测对HIV传播阻断非常重要。文章对HIV早期感染检测技术及相应技术,在急性HIV感染诊断、血液筛查及发病率估算中的使用情况及策略做一综述。新技术的发展和改进是HIV早期感染检测策略发展的关键。     

12例急性期HIV感染者抗病毒治疗结果分析

目的初步探讨急性期艾滋病病毒(HIV)感染者进行抗病毒治疗(ART)对其预后的影响。方法对在佑安医院接受抗病毒治疗的12例急性期HIV感染者进行跟踪随访和分析。结果 12例HIV感染者均在感染6个月内开始ARV治疗,并坚持服药未中断治疗,随访治疗期6个月-3年。治疗后3个月内检测病毒载量均低于最低检测限,1年后CD4+T细胞数均>500/μL,无明显严重药物不良反应发生,HIV感染者生存状态良好。结论初步结果显示,急性期开始抗病毒治疗可能为艾滋病病人带来好的预后结果。     

核酸定量检测试验应用于HIV-1感染诊断的评价

正1型艾滋病病毒(HIV-1)感染的诊断依赖于实验室检测,不同的检测方法由于标志物的不同而有不同的窗口期。常规的酶联免疫吸附试验(ELISA)+蛋白印迹试验(WB)的检测策略存在早期感染漏检的风险。在可疑人群中应用更高效的检测策略,可以有效地诊断HIV-1的早期感染。对2018年本确证实验室复检的第四代HIV筛查试剂有反应而WB结果为阴性或不确定的样本进行HIV-1核酸定量检     

绍兴市HIV-1毒株的耐药变异研究

目的了解绍兴市1型艾滋病病毒(HIV-1)株的耐药变异情况。方法分析2013-2014年绍兴地区新确认未经抗病毒治疗的130例HIV-1感染者,以及2014年前抗病毒治疗满一年且病毒未抑制的34例感染者样本,采用反转录-聚合酶链式反应(RT-PCR)和巢式-聚合酶链式反应(Nested-PCR)方法扩增HIV-1的pol基因区全长,参照美国斯坦福大学HIV耐药数据库,确定HIV亚型及耐药突变位点。结果 34例新诊断HIV感染者及33例病毒未抑制感染者的样品检测到耐药突变,主要亚型为CRF01_AE和CRF07_BC。仅在病毒未抑制感染者中检测到针对核苷类反转录酶抑制剂(NRTIs)的耐药突变,主要类型有M184V(42.42%)、K70R(15.15%)、D67N(12.12%)。在新确认感染者和病毒未抑制感染者两组人群中,均检测到针对非核苷类反转录酶抑制剂(NNRTIs)耐药的突变,主要类型有V179D/E/T(10.00%和18.18%)、K103N(2.22%和21.21%);在病毒未抑制感染者中还有Y181CY(21.21%)、V90I(12.12%)、G190A(15.15%)。在反转录酶(RT)区和整合酶(IN)区仅检测到少量次要耐药突变。结论绍兴市HIV经抗病毒治疗后易产生耐药性,部分可在新感染人群中传播,应对HIV耐药变异加强监测。     

孕期与产时启动抗病毒治疗阻断艾滋病母婴传播效果评价

目的对比艾滋病病毒(HIV)感染的孕产妇,经孕期与产时不同时期启动抗反转录病毒治疗(ART)后艾滋病母婴阻断的效果,探讨最大程度地减少通过母婴传播导致儿童艾滋病感染的方法。方法以30对临产时才发现HIV感染并采取紧急方案ART的孕产妇及其婴儿为观察组,以90对孕期开始应用ART的HIV感染孕产妇及其婴儿为对照组,两组均结合适宜的助产服务和人工喂养,婴儿进行HIV-1核酸和HIV抗体检测,判断HIV感染情况并进行对比分析。结果观察组有1例婴儿出生3个月死亡,有2例婴儿2次HIV-1脱氧核糖核酸检测结果均阳性,诊断婴儿HIV感染,其余27例满12~18月龄婴儿HIV抗体检测结果阴性,校正后婴儿HIV感染率为8.17%,对照组90例婴儿早期诊断和满12~18月龄HIV抗体检测结果均为阴性,婴儿HIV感染率为0,两组比较差异有统计学意义(χ~2=6.102,P0.05)。结论 HIV感染的孕产妇抗病毒治疗可以降低母婴传播的风险,孕期开始启动ART预防艾滋病母婴传播的效果明显优于产时紧急ART,结合安全分娩和人工喂养可明显降低HIV母婴传播率。     

提高对HIV-1急性感染和HIV-2诊断能力的检测策略北大核心

针对多年使用的艾滋病病毒(HIV)检测策略存在的问题,以及目前的检测技术研发和应用状况,美国疾病控制和预防中心在2014年6月提出了新的HIV检测策略,联合使用HIV抗原和抗体检测试剂、能够区分HIV-1和HIV-2抗体的诊断试剂以及HIV-1核酸定性诊断试剂,提高对HIV-1急性感染和HIV-2的检出效能,同时减少需要随访的不确定结果,缩短等待时间。文章对这个检测策略进行了全面介绍,并提出制定适合我国的HIV检测策略的建议。     

新疆伊宁市912例HIV-1型病毒载量定量检测结果分析

目的分析新疆伊宁市人民医院抗病毒中心912例HIV感染者HIV-1型病毒载量定量检测结果,了解HIV感染者体内病毒载量、病毒载量与临床指征之间的相互关系,为艾滋病的预防和抗病毒治疗提供基础数据。方法采用分枝DNA信号放大系统(bDNA)技术,利用SIEMENS公司VERSANT-440分子系统,2012年7月～2013年2月检测伊宁市912例HIV感染者HIV-1型病毒载量拷贝数。结果伊宁市912例HIV感染者HIV-1病毒载量分布状况:<500 c/ml 661例(72.4%),501～3 000 c/ml 48例(5.3%),3 001～10 000 c/ml 82例(9.0%),10 001～30 000 c/ml 49例(5.4%),30 000～3 5000 c/ml 6例(0.7%),>35 000 c/ml 66例(7.2%)。结论 HIV-1型病毒载量定量检测是判定HIV感染者疾病进程,评估临床抗病毒治疗的重要指标。     

抗结核治疗对HIV/AIDS-TB病人抗病毒治疗效果的影响

目的分析艾滋病病毒(HIV)感染者/艾滋病(AIDS)病人合并肺结核(TB)(简称HIV/AIDS-TB病人)抗结核治疗(ATT)对抗病毒治疗(ART)效果的影响因素和HIV-1基因型耐药情况。方法回顾性收集2014年1月至2017年12月云南省传染病医院确诊的HIV/AIDS-TB病人和HIV/AIDS病人的人口学、临床资料、HIV-1病毒载量和基因型耐药,分析影响HIV/AIDS-TB病人HIV-1复制的因素。结果 197例HIV/AIDS-TB病人在联合治疗24周后HIV核糖核酸(RNA)检测阳性率为23.86%(47例),抗病毒治疗失败率为11.17%(22例),抗病毒失败病人的耐药率为9/22,均高于ART的HIV/AIDS病人,但二者间的差异均无统计学意义。HIV/AIDS-TB病人主要核苷类反转录酶抑制剂相关的突变为M184V(26.67%),主要非核苷类反转录酶抑制剂相关的突变为V179D(26.67%)和V106M(20.00%),未产生蛋白酶抑制剂主要突变。二分类多因素Logistic回归分析发现,入组时的白蛋白(ALB)、抗酸杆菌涂片、CD4+T淋巴细胞、CD8+T淋巴细胞、CD4/CD8+T淋巴细胞比值、ART和ATT方案与HIV-1复制无关,而ATT 8周内接受ART是抑制HIV/AIDS-TB病人HIV-1复制的保护因素。结论联合治疗能有效抑制HIV/AIDS-TB病人的HIV-1病毒载量,且未加速病人HIV-1基因型耐药的产生。对于尚未接受ART的双重感染病人在启动抗结核治疗8周内开始ART有利于抑制HIV-1病毒载量。     

Time of initiation of antiretroviral therapy: impact on HIV-1 viraemia. The Swiss HIV Cohort Study

OBJECTIVE: The current recommendation that patients infected with HIV-1 be treated early is based on little evidence. We examined whether the early initiation of antiretroviral treatment affects residual HIV-1 viraemia. METHODS: Viraemia was measured using an assay with a detection limit of 3 HIV-1 RNA copies/ml in drug-naive patients who started antiretroviral therapy at the time of primary HIV-1 infection (PHI) (n = 10), during chronic infection without immune suppression (CD4 cell counts > or = 500/mm3; median 577) (n = 10), or after immune suppression developed (CD4 cell counts < 500/mm3; median 113) (n = 21). RESULTS: In 249 samples collected 24 to 120 weeks after treatment initiation, the mean proportion of samples with HIV-1 RNA levels of less than 3 copies/ml was 75% for PHI patients compared with 32 and 8% for immunocompetent and immunosuppressed chronically infected patients, respectively. Fifty per cent of PHI patients, but none of the chronically infected patients, had persistently fewer than 3 HIV-1 RNA copies/mL. PHI patients had lower residual HIV-1 RNA levels than chronically infected patients, and immunocompetent patients had lower residual HIV-1 RNA levels than immunosuppressed patients (all pairwise, P< 0.001). The mean residual HIV-1 RNA level was independently associated with the initiation of therapy during PHI and baseline CD4 cell counts (P < 0.001 for both associations). CONCLUSION: Viraemia levels are associated with clinical progression and predict virological treatment failure. The initiation of antiretroviral therapy at the time of PHI and while CD4 cell counts are high results in lower residual viraemia. These results support early antiretroviral therapy in HIV-1-infected patients.     

Early induction of leukemia inhibitor factor (LIF) in acute HIV-1 infection

BACKGROUND: Leukemia inhibitor factor (LIF) is thought to play a substantial role in protecting CD4 T cells in lymphoid tissues (LT) from infection by HIV-1. OBJECTIVE: To investigate whether primary HIV-1 infected subjects with sustained virological control (< 1000 HIV-1 RNA copies/ml plasma) post-cessation antiretroviral therapy (ART) had a higher initial LIF response during primary HIV-1 infection (PHI) as compared with those individuals who did not achieve a similar control (> 9000 HIV-1 RNA copies/ml plasma) of HIV-1 replication. MATERIAL AND METHODS: Consecutively obtained HIV plasma samples were collected from primary HIV-1 infected subjects. A group of acutely Epstein-Barr virus-infected subjects and a group of HIV-1-seronegative healthy individuals served as controls. All samples were tested by ELISA for LIF and sgp130, the soluble form of LIF's signalling receptor. RESULTS: LIF and sgp130 plasma levels were significantly increased in primary HIV-1-infected subjects as compared with HIV-1-seronegative controls. Peak plasma levels of LIF occurred during the first week of PHI whereas sgp130 peaked between 2 and 4 weeks after the onset of PHI. Furthermore a positive correlation was found between viral load and plasma levels of LIF during PHI. Both LIF and sgp130 plasma concentrations were significantly lower during the viral rebound phase after treatment interruption as compared with the PHI phase. CONCLUSIONS: LIF induction occurred in the initial stages of viral dissemination during PHI. It may be a part of the virally induced generalized pro-inflammatory response. LIF levels at PHI did not predict low levels of HIV viraemia after discontinuation of ART. LIF was not increased following ART interruption in this early treated population.     

基于核酸扩增方法估计MSM的HIV新发感染的研究

目的采用核酸扩增方法（NAAT）,估计男男性行为人群（MSM）急性艾滋病病毒（HIV）感染（AHI）的新发感染情况。方法采集MSM抗凝和非抗凝全血各10mL,非抗凝血分离血清检测HIV抗体。HIV抗体阴性的MSM抗凝全血分离血浆,进行HIV1核糖核酸（RNA）混合NAAT检测。NAAT检测阳性样本计算新发感染。结果800名调查对象中,酶联免疫吸附试验（ELISA）检测HIV抗体阳性64人,经蛋白印迹试验（wB）检测均确定为HIV1感染,阳性检出率8．00％E95％可信区间（CI）：6．04-9．96）。ELISA检测HIv抗体阴性的736人,经过HIV-1RANNAAT检测,检出5例阳性。依据公式计算,AHI检出率为0．68％（5／736）。每年新发感染率为8．86％（95％CI：1．07～16．65）。结论NAAT是能有效检测出HIV新发感染的方法,可以减少血清学试验的漏检,对防控艾滋病的扩散具有一定的临床意义。     

集合PCR在检测MSMHIV急性期感染中的应用

目的应用集合聚合酶链反应（PCR）方法,发现广州市男男性行为人群（MSM）中艾滋病病毒（HIV）急性感染者,并估计该人群HIV感染的发病率。方法采用10∶1、5∶1、1∶1三级集合HIV-1核糖核酸（RNA）反转录PCR（RT-PCR）方法进行检测。结果 2008年,检测1 250份MSM HIV-1抗体阴性血浆,发现HIV-1RNA阳性3例,追踪随访,2例HIV-1抗体阳转,1例失访;初步估计HIV-1年发病率约为8.50%[95%可信区间（CI）：1.75%～24.86%]。2009年,检测1 002份MSM HIV-1抗体阴性血浆,发现HIV-1RNA阳性4例,追踪随访,4例均HIV-1抗体阳转;估计HIV-1年发病率约为14.17%（95%CI：3.85%～36.21%）。结论集合HIV-1RNART-PCR是发现急性感染者的有效方法,并可估计人群HIV发病率。     

Rapid restoration of CD4 T cell subsets in subjects receiving antiretroviral therapy during primary HIV-1 infection

OBJECTIVE: To compare the effect of highly active antiretroviral therapy on immune reconstitution in subjects with acute and chronic HIV-1 infection. DESIGN: Prospective study including 58 treatment-naive subjects who commenced indinavir or nelfinavir and two nucleosides during primary (PHI; n = 28) or chronic HIV-1 infection (CHI; n = 30). METHODS: Naive (CD45RA+ 62L+), memory (CD45RA-) and activated (CD38+ HLA-DR+) T cell subsets were quantified at 1-2 monthly time intervals using 4-colour flow cytometry. RESULTS: At 1 year, HIV-1 RNA declined in both cohorts to undetectable levels (< 50 copies/ml), while median CD4 lymphocyte count increased from 470 to 758 x 10(6) cells/l in PHI and from 204 to 310 x 10(6) cells/l in CHI, reaching > 500 x 10(6) cells/l in 93% of PHI, but only in 37% of CHI subjects (P < 0.001). Naive CD4 lymphocytes increased from 106 to 176 x 10(6) cells/l in PHI and from 41 to 44 x 10(6) cells/l in CHI (PHI versus CHI at 12 months: P = 0.003), while memory cells rose from 368 to 573 x 10(6) cells/l in PHI and from 148 to 223 x 10(6) cells/l in CHI (P < 0.001). Early increases (< 3 months) of CD4 lymphocytes were larger in subjects with PHI, consisting of naive CD45RA+ CD62L+ as well as memory CD45RA- CD62L+ cells (P = 0.001). CD4 activation declined from 5 to 2% in PHI and from 13 to 6% in CHI (P = 0.001), while CD8 cell activation was reduced from 33 to 15% in PHI and from 42 to 19% in CHI (P = 0.02). CONCLUSION: Immune reconstitution was more complete, occurred earlier and comprised both naive and memory CD4 T lymphocytes in subjects who commenced antiretroviral therapy during primary HIV-1 infection.     

Increased turnover of CCR5+ and redistribution of CCR5- CD4 T lymphocytes during primary human immunodeficiency virus type 1 infection

CCR5 is the major coreceptor for human immunodeficiency virus (HIV) type 1 during primary infection. CCR5+ CD4 T lymphocytes were studied in subjects with primary HIV-1 infection (PHI) or acute Epstein-Barr virus (EBV) infection and in HIV-uninfected controls. The early decline of CD4 T lymphocytes during PHI resulted from depletion of CCR5- CD4 T lymphocytes. After antiretroviral therapy, Ki-67- CCR5- CD4 T cell counts rapidly increased in the circulation, which suggests that the initial decrease was due to an alteration in trafficking and/or sequestration. In the CCR5+ subset of CD4 T cells, there was an elevation in the proliferative (Ki-67+) fraction during PHI, yet their total number remained in the normal range. In contrast, in acute EBV infection, proliferating CCR5+ CD4 T cells accumulated to very high levels, suggesting they have an important role in the early antiviral response, which may be impaired in HIV-1 infection.     

Suppression of leukemia inhibitor factor in lymphoid tissue in primary HIV infection: absence of HIV replication in gp130-positive cells

BACKGROUND: Leukemia inhibitor factor (LIF), a member of the interleukin-6 cytokine family, has recently been shown to inhibit HIV-1 replication both in vivo and in vitro. OBJECTIVE: LIF and its corresponding receptors gp130 and LIF receptor-alpha (LIFR-alpha) were studied in lymphoid tissue (LT) to reveal potential systemic immunoregulatory effects during the course of HIV-1 infection. METHODS: LIF, gp130, LIFR-alpha and HIV-1 replicating cells were identified at the single cell level by immunohistochemistry and quantified by computerized in situ imaging in tonsil and lymph nodes biopsies (LT) from patients with primary HIV-1 infection (PHI), chronic HIV-1 infection (cHI), long-term non-progressors (LTNP) and HIV-1 seronegative controls. RESULTS: LIF and its receptors, gp130 and LIFR-alpha were significantly (P < 0.005) upregulated in LT from PHI patients as compared with HIV-1 seronegative controls. Expression of LIF in cHI was comparable to LIF levels in HIV-1 seronegative controls whereas LTNP showed significantly reduced LIF expression (P < 0.05). LIF receptors, gp130 and LIFR-alpha were significantly upregulated in cHI (P < 0.005) but downregulated in LTNP (P < 0.05 and P < 0.005, respectively). LIF expressing cells could be demonstrated in LT 2 days after onset of acute retroviral syndrome. LIF expression was evident in CD3, CD4 and CD8 cells. Furthermore, high plasma viral load was associated with high expression of LIF in LT. Finally, no HIV-1 replication could be found in CD4 gp130-positive cells in PHI. CONCLUSIONS: LIF, gp130 and LIFR-alpha showed increased expression in LT from patients with PHI. Furthermore, HIV-1 replication did not occur in cells expressing the LIF signaling receptor, gp130, indicating that LIF may be associated with control of HIV-1 replicating cells in vivo.     

Potent antiretroviral therapy of primary human immunodeficiency virus type 1 (HIV-1) infection: partial normalization of T lymphocyte subsets and limited reduction of HIV-1 DNA despite clearance of plasma viremia.

Antiretroviral therapy commenced during primary human immunodeficiency virus type 1 (HIV-1) infection (PHI) may limit the extent of viral replication and prevent early loss of HIV-specific CD4 lymphocyte function. We studied the effect of current standard therapy (2 nucleoside analogues and a protease inhibitor) in 16 patients with symptomatic PHI. In the 13 patients who completed 1 year of treatment, plasma HIV RNA was <50 copies/mL and median CD4 cell counts were comparable to HIV-uninfected controls, with naive (CD45RA+CD62L+), primed (CD45RO+), and T cell receptor Vbeta subsets all within normal ranges. However, HIV-1 DNA levels in treated and untreated PHI patients were similar. Furthermore, CD8 cell counts remained elevated, including activated (CD38+HLA-DR+), replicating (Ki-67+), and cytotoxic (perforin+CD28-) lymphocytes. In conclusion, early antiretroviral therapy resulted in clearance of viremia and prevented loss of crucial CD4 subsets. The persistence of HIV-1 DNA together with increased CD8 T lymphocyte turnover and activation indicate continued expression of viral antigens.