Molecular epidemiology of the heterosexual HIV-1 transmission in Kunming, Yunnan Province of China suggests origin from the local IDU epidemic

Molecular epidemiological investigation was conducted among injecting drug users (IDUs) (n = 11) and heterosexuals (n = 15) in Kunming, Yunnan Province of China. HIV-1 genotypes were determined based on the nucleotide sequences of 2.6-kb gag-RT region. The distribution of genotypes among IDUs was as follows: CRF07_BC (5/11) and CRF08_BC (5/11); subtype B' (1/11). Similarly, a majority of Kunming heterosexuals (14/15) were infected with CRF07_BC (4/15), CRF08_BC (6/15), or subtype B' (4/15), known to predominate among IDUs in China. This contrasts with trends in the coastal regions of China and surrounding southeastern Asian countries, where CRF01_AE predominates among heterosexuals. The heterosexual HIV-1 epidemic in Kunming thus appears to derive from the local IDU epidemic. Of note, subtype B' was the most prevalent strain among heterosexuals before 1997, while CRF07_BC and CRF08_BC became predominant in 2002, indicating a transition of HIV-1 genotype distribution between the early and the more recent samples from Kunming heterosexuals.

In vitro anti-HIV and -HSV activity and safety of sodium rutin sulfate as a microbicide candidate

Sodium rutin sulfate (SRS) is a sulfated rutin modified from the natural flavonol glycoside rutin. Here, we investigated its in vitro anti-HIV and -HSV activities and its cytotoxic profile. Fifty percent inhibitory concentration (IC50) values of SRS against HIV-1 X4 virus IIIB, HIV-1 R5 isolates Ada-M and Ba-L were 2.3 +/- 0.2, 4.5 +/- 2.0 and 8.5 +/- 3.8 mu M with a selectivity index (SI) of 563, 575 and 329, respectively. Its IC50 against primary R5 HIV-1 isolate from Yunnan province in China was 13.1 +/- 5.5 mu M, with a Sl of 197. In contrast, unsulfated rutin had no activity against any of the HIV-1 isolates tested. Further study indicated that SRS blocked viral entry and virus-cell fusion likely through interacting with the HIV- I envelope glycoprotein. SRS also demonstrated some activity against human herpes simplex virus (HSV) with an IC50 of 88.3 +/- 0.1 mu M and a Sl of 30. The 50% cytotoxicity concentration (CC50) of SRS was >3.0 mM, as determined in human genital ME 180, HeLa and primary human foreskin fibroblast cells. Minimum inhibitory concentration of SRS for vaginal lactobacilli was >3.0 mM. These results collectively indicate that SRS represents a novel candidate for anti-HIV-1/HSV microbicide development. (C) 2007 Elsevier B.V. All rights reserved.

HIV-1 C亚型密码子优化gp120负载人DC疫苗的构建及体外功能

目的构建HIV-1C亚型gp120负载人树突状细胞(dentriti ccell,DC)疫苗,并对其体外功能进行初步检测。方法利用Amaxa细胞核转染技术将pcDNA3.1-gp120质粒转染至人成熟DC,以Western blot检测gp120的表达。通过流式细胞仪检测DC表面共刺激分子的变化、混合淋巴细胞反应、CD8+T细胞表面活化分子CD25的表达及其分泌IFN-γ的变化。结果通过Western blot检测,gp120在DC中得到了正确表达。经流式细胞仪检测,DC表面分子CD80表达率由刺激前的33.34%上升至43.20%,CD86表达率由刺激前的60.08%上升至90.34%;负载gp120DC刺激淋巴细胞增殖率为86.72%;CD8+T细胞表面分子CD25表达率由刺激前的5.27%上升至74.21%,IFN-γ的表达率达37%。结论负载了HIV-1gp120的人树突状细胞能够显著刺激淋巴细胞的增殖、增强CD8+T细胞表面活化分子CD25表达以及促进CD8+T细胞分泌IFN-γ,为下一步DC治疗性疫苗的体内研究奠定基础。

基于Vif-APOBEC3G相互作用的抗HIV-1药物研究

载脂蛋白B mRNA编辑酶催化多肽样蛋白3G(apolipoprotein B mRNA-editing enzyme catalyticpolypeptidelike3G,APOBEC3G或A3G)是人体天然抗病毒分子,可以使病毒逆转录形成的cDNA的胞嘧啶(C)脱氨为尿嘧啶(U),产生鸟嘌呤(G)→腺嘌呤(A)超突变,导致病毒转录产物突变,从而达到抑制病毒复制的作用。HIV-1的辅助蛋白Vif,可与APOBEC3G相互作用并导致其被降解,使得这一天然抗病毒机制失效,进而增强了HIV的感染力。Vif与APOBEC3G这种相互作用为抗HIV药物提供了新靶点。针对Vif-APOBEC3G相互作用的抗HIV抑制剂已经成为研究热点。本文综述了Vif和APOBEC3G的结构、二者的相互作用,以及基于这一相互作用的抗HIV-1抑制剂研究进展。

6-(1H-吲哚-3-甲基)-5-乙基-3H-嘧啶-4-酮类化合物的合成及抗HIV活性研究

基于二氢烷氧苄基嘧啶酮(DABO)类非核苷类逆转录酶抑制剂(NNRTIs)的构效关系研究,设计合成了2个新的6-(1H-吲哚-3-甲基)-5-乙基-3H-嘧啶-4-酮类化合物,并采用C8166细胞进行了体外抗HIV活性测试,为新型S-DABO类非核苷类逆转录酶抑制剂结构修饰提出了新的设想.

天花粉蛋白抗HIV．1构效关系及机制探讨

目的研究TCS抗HIV-1的构效关系并对机制进行探讨。方法蛋白工程技术构建14个TCS突变 体，测定各种TCS突变体的细胞毒性和抗HIV恬性。结果活性中心突变体TCSM(120—123)与TcSEl60A／E189A在失 去绝大部分RI活性的同时，也几乎完全失去抗HIV活性。而另一个活性中心突变体TCSRl22G，RI活性下降160 倍，却仍保留一定的抗HIV活性。TCSC末端删除突变体(TCS(2，TCSo和TCScl。)抗HIV活性的下降(1．4—4．8 倍)与其RI活性呈平行下降(1．2—3．3倍)；分别在C末端加上末端19个氨基酸延伸肽或KDEL信号肽的突变体 TCscl9。与TQkDEL，虽然保留全部的RI活性，但却几乎完全失去抗HIV活性；TCS抗原决定簇位点突变后对TCS 抗HIV．1括性没有显著影响，但当在抗原决定簇突变体所引入的Cys残基上加上PEG20K后，这些突变体则显著降 低了抗HIV-1的活性。TCS不能抑制HIV一1进入宿主细胞；对感染细胞和未感染细胞的融合没有抑制作用；TCS 也不能抑制HIV一1 rRT活性；TCS对病毒颗粒的直接杀伤作用不大。结论TCS抗HIV-1活性与其Rl活性显著 相关，但似乎又不是唯一的决定因素；TCS C末端氨基酸的突变影响其抗HIV．1活性；在C末端增加KDEL信号肽 序列及19aa尾肽并不能增加其抗HIV活性；抗原决定簇突变体以及PEG20K偶联TCS抗原决定簇突变体体外抗 HIV-1活性下降；抗HIV．1机制可能与对感染细胞的间接作用有关。

AIDS猕猴模型在HIV疫苗研究中的应用

：对HIV 疫苗的研究一直是国际上艾滋病方面研究的热点和难点。动物模型则为疫苗研究必不可缺少的 重要工具，缺乏合适的动物模型很大程度上制约了AIDS疫苗的研究。目前在国际上SIV 或SHIV 感染的猕猴模 型为最常用的AIDS研究模型，受猕猴背景及病毒特性等多种因素的影响，使得以上两种模型在HIV疫苗研究中 仍存在一定的局限性。为了更好地发挥猕猴模型在HIV疫苗研究中的巨大潜力，开发理想的AIDS猕猴模型已成 为目前HIV疫苗研究的首要任务。本文简要介绍了AIDS疫苗的研发策略、研发概况以及SIV／SHIV猕猴模型在 HIV疫苗中的应用，并对其中存在的问题及其应用前景进行了探讨。

纳米银体外抗HIV 活性的研究

［ 目的］ 制备纳米银颗粒并检测其抗人类免疫缺陷病毒（HIV） 活性及其体内外毒性， 探讨纳米银体外抗 HIV 的作用机制。［ 方法］ 在体外培养C8166 细胞中加入不同浓度的纳米银， 检测其体外细胞毒性CC50； 不同病毒株 感染细胞， 同时加入不同浓度的纳米银， 孵育后检测细胞病变及病毒抗原， 检测纳米银对HIV 的抑制作用。纳米银滴鼻 处理小鼠， 检测小鼠体重变化， 并进行病理切片， 观察纳米银滴鼻处理小鼠的肝、肾及肺组织的病理变化。［ 结果］ 纳米银颗粒对C8166 细胞毒性的CC50 为105.73 μg ／ ml， 抑制HIV 实验株HIV-1IIIB， HIV-2CBL-20 诱导细胞病变的 EC50 分别为26.56 μg ／ ml 和35.47 μg ／ ml； 抑制HIV-1IIIB p24 抗原产生的EC50 为9.80 μg ／ ml； 直接杀病毒作用的EC50 为 12.08 μg ／ ml。纳米银经滴鼻处理的小鼠， 其体重及肝、肾及肺组织切片与对照组比较， 均无明显改变。［ 结论］ 纳米 银有较好的抗HIV 活性， 作用机制可能主要通过直接杀病毒来抑制HIV 的复制； 纳米银在呼吸道黏膜局部应用无明显 的毒副作用， 作为黏膜表面抗病毒药物， 值得进一步深入研究。

缅甸HIV分子流行病学及与我国HIV流行的相关性

缅甸与我同毗邻，中缅困境线长，人员交往密切，缅甸HlV毒株可以通过各种途径传人 我国，有可能成为我国新型HIV毒株传播和流行的来源，使我国HIV病毒亚捌的流行更加复杂多样。 通过加强埘缅甸HIV感染人群的病毒弧型的分析研究，将有利于监测我闷HIV流行株的分布和流行 特点，从而采取有效的应对措施，为防止新的HIV亚型和重组型在我国的传播和扩大流行做好积极 的准备工作。

A switch-on mechanism to activate maize ribosome-inactivating protein for targeting HIV-infected cells

Maize ribosome-inactivating protein (RIP) is a plant toxin that inactivates eukaryotic ribosomes by depurinating a specific adenine residue at the a-sarcin/ricin loop of 28S rRNA. Maize RIP is first produced as a proenzyme with a 25-amino acid internal inactivation region on the protein surface. During germination, proteolytic removal of this internal inactivation region generates the active heterodimeric maize RIP with full N-glycosidase activity. This naturally occurring switch-on mechanism provides an opportunity for targeting the cytotoxin to pathogen-infected cells. Here, we report the addition of HIV-1 protease recognition sequences to the internal inactivation region and the activation of the maize RIP variants by HIV-1 protease in vitro and in HIV-infected cells. Among the variants generated, two were cleaved efficiently by HIV-1 protease. The HIV-1 protease-activated variants showed enhanced N-glycosidase activity in vivo as compared to their un-activated counterparts. They also possessed potent inhibitory effect on p24 antigen production in human T cells infected by two HIV-1 strains. This switch-on strategy for activating the enzymatic activity of maize RIP in target cells provides a platform for combating pathogens with a specific protease.

APOBEC3G抗HIV一1作用机制研究进展

】APOBEC3G是细胞内新的广谱抗病毒蛋白。它具有胞苷脱氨酶活性，能使病毒负链 DNA产生dC--·dU高发突变，造成正链DNA G---A突变，使得病毒DNA变成无功能或降解。APO— BEC3G具有广泛的生物学功能，可以限制HIV-1等病毒复制，但HIV-1病毒感染因子Vii能拮抗 APOBEC3G抑制病毒活性作用。本文简要综述了APOBEC3G抑制病毒HIV-1作用机制的最新进 展。

Recombinant protein of heptad-repeat HR212, a stable fusion inhibitor with potent anti-HIV action in vitro

HR212, a recombinant protein expressed in Escherichia coli, has been previously reported to inhibit HIV-1 membrane fusion at low nanomolar level. Here we report that HR212 is effective in blocking laboratory strain HIV-1IIIB entry and replication with EC50 values of 3.92±0.62 and 6.59±1.74 nM, respectively, and inhibiting infection by clinic isolate HIV-1KM018 with EC50 values of 44.44±10.20 nM, as well as suppressing HIV-1- induced cytopathic effect with an EC50 value of 3.04±1.20 nM. It also inhibited HIV-2ROD and HIV-2CBL-20 entry and replication in the μM range. Notably, HR212 was highly effective against T20-resistant strains with EC50 values ranging from 5.09 to 7.75 nM. Unlike T20, HR212 showed stability sufficient to inhibit syncytia formation in a time-of-addition assay, and was insensitive to proteinase K digestion. These results suggest that HR212 has great potential to be further developed as novel HIV-1 fusion inhibitor for treatment of HIV/ AIDS patients, particularly for those infected by T20-resistant variants.