柳州和南宁HIV-1亚型分布及耐药情况调查

为了解柳州和南宁两市HIV-1亚型分布和耐药情况,在柳州和南宁招募HIV感染者和AIDS患者共304名,采集外周静脉血,从血浆中提取HIVRNA,扩增HIVpol基因并测序。将获得的序列进行系统进化树分析,结果表明柳州的HIV-1毒株中存在CRF01_AE和CRF07_BC两种亚型,其中CRF01_AE毒株占75.2%,CRF07_BC毒株占24.8%;南宁的HIV-1毒株中存在CRF01_AE、CRF08_BC、B亚型和C亚型共4种亚型,其中CRF01_AE和CRF08_BC仍是南宁最主要的亚型,CRF01_AE占85.8%,CRF08_BC占11.5%。根据所得的序列资料进行HIV-1耐药性分析,计算耐药率。计算结果表明,柳州未治疗和治疗研究对象的耐药率分别为3.3%和8.7%,南宁未治疗和治疗研究对象的耐药率分别为1.4%和27.5%。     

江苏省2017年新报告HIV感染者治疗前耐药分析及基因亚型分布

基于大样本量的亚型及治疗前耐药分析,更准确地掌握江苏省HIV-1流行亚型,更真实地评估江苏省治疗前耐药水平并为抗病毒治疗的开展提供依据收集2017年全年全江苏省新报告感染者治疗前血样.逆转录PCR扩增HIV-1蛋白酶区(pol)基因并送测序.ChromasPro剪辑,拼接,合成序列.MEGA7做序列比对并构建进化树分析亚型.序列在线比对耐药位点.治疗前耐药分析,耐药率达16.3％.蛋白酶区(PR)主要耐药位点是L33F,占44.4％.核苷酸逆转录酶区(NRTI)主要耐药位点是M184位点变异,占35％.非核苷酸逆转录酶区(NNRTI)耐药位点主要是V179位点变异,占59.2％.CRF01_AE亚型(456,37.7％)和CRF07_BC亚型(381,31.6％)仍为江苏省主要流行亚型;不同的是复杂重组体(CPXs)和独特重组体(URFs)数量极速上升至第三位(241,19.9％).江苏省HIV-1复杂重组体大量出现,提示新人病毒或病毒交叉重组频繁,艾滋病的防制依然任重道远.在治疗前耐药比例大幅上升的情况下,治疗前耐药检测成为个体选择抗病毒治疗方案的关键.     

上海地区HIV毒株基因亚型分布及原发基因耐药的分子流行病学研究

本文通过对未经抗病毒治疗患者的人类免疫缺陷病毒1型（HIV-1）毒株进行检测,了解上海地区HIV-1的亚型分布及原发耐药基因变异现状。对118例未经治疗的HIV感染者其标本中HIV蛋白酶全长和部分反转录酶基因进行反转录-聚合酶链反应（RT-PCR）扩增,经DNA测序后进行系统进化树分析和重组分析,以确定HIV-1基因亚型和重组体,并与斯坦福耐药数据库比对,了解耐药性突变位点。使用斯坦福REGA HIV亚型分型工具和美国国立生物技术信息中心（NCBI）HIV亚型分析工具分析亚型,获得118例患者的HIV基因序列,基因分型分别为CRF01_AE重组体57例(48.3%)、B亚型36例(30.5%)、CRF07_BC 15例 (12.7%)、CRF08_BC 7例(5.9%)、C亚型2例(1.7%),亚型间或重组体间二重重组体(B/CRF01_A E)1例(0.8%)。蛋白酶抑制剂(PI)和反转录酶抑制剂相关的耐药基因突变率达54.2%（64/118）,其中2例（1.7%）发生PI耐药,基因突变位点：M46L、Q58E。5例(4.1%)对反转录酶抑制剂产生耐药,其中对核苷类反转录酶抑制剂（NRTI）和非核苷类反转录酶抑制剂（NNRTI）的耐药率分别为3例(2.4%)和5例(4.1%)。基因突变位点：NRTI为M41L、D67N、T69I/N/S、K70L、L74V、V75L、V118I、M184V、L210W/F/M/S和T215F;NNRTI为V90I、L100V、K103R/N、V106M/P/I/G、E138G/A、V179E/D/T、Y181C、G190A、H221Y、F227L、K238S和Y318F。结果提示,上海地区HIV毒株以CRF01_AE重组亚型为主,且发现新的重组体,可能出现新重组体流行的趋势。PI和反转录酶抑制剂相关的耐药基因突变率较高,且存在高度原发耐药毒株,应加强HIV-1耐药基因变异监测,科学、合理地给予抗病毒治疗。     

2020年银川市HIV-1感染者基因亚型及耐药分析

为了解银川市接受抗病毒治疗的HIV-1感染者基因亚型和耐药突变特征。收集2020年银川市接受抗病毒治疗的HIV-1感染者血样,其中病毒载量大于400拷贝/mL的样本进行基因型耐药检测,通过巢式聚合酶链反应（PCR）扩增HIV-1 pol基因区片段并测序,对所得序列进行基因分型和耐药突变分析。收集642例血浆样本,50例病毒载量大于400拷贝/mL,基因型耐药检测后获得29例pol区基因序列。17例耐药,3例潜在耐药,总体耐药率为2.65%(17/642)。CRF07＿BC和CRF01＿AE是耐药株的主要基因亚型。NRTIs类耐药突变中,M184V/I(31.03%)突变发生率最高,其次为K65R(17.24%)。3TC、FTC和ABC耐药发生率最高,均为37.93%(11/29)。NNRTIs类耐药突变中,G190A/S/E、V179D/DE/E突变发生率最高,均为24.14%;K103N/KN(17.24%)次之。EFV和NVP耐药发生率最高,均为58.62%(17/29)。2020年银川市新型重组亚型增加,首次检出CRF79＿0107。总体耐药率处于低水平,以NNRTIs耐药为主,...     

上海地区人类免疫缺陷病毒1型感染/艾滋病患者病毒耐药基因突变及亚型分布

目的 研究上海地区人类免疫缺陷病毒1型(HIV-1)感染／艾滋病(AIDS)患者中HIV-1耐药株出现的情况及亚型分布。方法 对33例HIV-1感染／AIDS患者的血浆HIV-1分离株,进行抗HIV-1药物(核苷类反转录酶抑制剂、非核苷类反转录酶抑制剂和蛋白酶抑制剂)的基因型耐药检测和亚型分析。结果 33例的HIV-1均未检出对PI的耐药突变;10例高效抗反转录病毒疗法(HAART)治疗失败或抑制病毒复制不完全者中,检出的耐药突变为70％,过渡型耐药突变为20％;23例未经抗HIV-1治疗者中,耐药突变为4．3％,过渡型耐药突变为13％。所有过渡型耐药突变均为T215S。15例经血制品传播的HIV- 1均为B亚型;18例经吸毒和性传播的HIV-1中,B和CRF01-AE亚型分别为39％,和33％,此外,还有C、D、G、K和CRF02-AG亚型。结论 上海地区HIV-1感染／AIDS患者中,HAART治疗失败或复制抑制不完全者HIV-1的NRTI和NNRTI耐药突变率高;吸毒和性传播者的HIV-1中,除主要为B和CRF01-AE亚型外,尚有其他少见的亚型。     

HIV-1 gag与gp41基因片段的序列特征与亚型研究

本文对华北地区出入境39例HIV-1阳性样本(中国21例,非洲17例,东南亚1例)的gag和env两个基因片段进行了序列特征和亚型对比分析。发现了A、A1、A3、B、C、G亚型和重组亚型03_AB、01_AE、AG、02_AG、07_BC、08_BC、CD和06_CPX共14个亚型,其中重组亚型占57.2%(8/14)。表明HIV-1基因变异较快,亚型分布广泛,重组亚型有增多趋势。此外发现26.7%(8/30)的样本,其gag和env基因区亚型表现不一致。提示在研究HIV-1亚型中应综合gag和env两个基因区的序列特征进行亚型分析。     

亚洲主要流行HIV-1 Gp120及其5个高变区多态性分析

HIV-1 Gp120及其5个高变区的多态性是HIV-1能够逃逸宿主免疫反应和耐药性的主要原因。下载了现有数据库中记载的包括B’、C、CRF01_AE、CRF07_BC和CRF08_BC 5种亚洲主要流行亚型和重组型的所有Gp120及其5个高变区序列,分析了HIV-1 Gp120及其5个高变区序列的长度多态性,糖基化位点数以及序列特征。结果显示,绝大部分的HIV-1 Gp120序列长度为496～515个氨基酸之间,这提示Gp120长度为496～515个氨基酸的HIV-1毒株可能是疫苗研究的理想毒株。比较5个高变区,V3区表现出了最低的长度多态性,几乎没有糖基化位点,表明受到较强的功能限制。不同亚型和重组型的V3区一致序列比较发现,所有的HIV-1亚型均表现为R5型。这提示阻断HIV-1病毒结合CCR5受体是治疗艾滋病的有效途径,R5型毒株可以作为艾滋病疫苗研究的理想毒株。另外,V1和V4区展现了很高的长度多态性,糖基化位点也较多;而V2和V5区的长度多态性和糖基化位点都较低。这些结果表明降低这4个高变区的多态性和糖基化位点数目可能解决HIV-1病毒对疫苗的逃逸,从而研究出真正有效的疫苗。     

Identification of a novel HIV-1 intra-circulating recombinant form 01_AE in China: a descendant of the previously identified CRF01_AE transmission clusters 1 and 6

<正>Dear Editors,Here,we report a novel HIV-1 intra-circulating recombinant form 01_AE(intra-CRF01_AE:CRF01-1AE/CRF01-6AE)composed of CRF01_AE transmission clusters 1 and 6,which was identified in a heterosexual male from Fujian,with one breakpoint observed in the vif gene.The CRF01_AE region I(HXB2:868–5184)of the recombinant clustered with CRF01_AE transmission cluster 1,which is     

表达HIV-1 CRF01＿AE亚型结构基因小鼠模型的建立和鉴定

目的:构建并鉴定表达HIV-1 CRF01_AE亚型结构基因的小鼠模型。方法:使用哺乳动物密码子优化的HIV-1 CRF01_AE gp160基因,通过慢病毒包装系统构建重组慢病毒LV-GFP-AE gp160,将上述重组慢病毒感染小鼠肺上皮细胞TC-1,经嘌呤霉素抗性筛选获得稳定表达gp160基因的TC-1细胞。采用RT-PCR、流式细胞术检测gp160基因在细胞内的表达稳定性,将稳定表达Gp160蛋白的TC-1-HIV AE gp160细胞接种小鼠,用免疫组化方法检测小鼠体内细胞团块中HIV Gp160蛋白的表达。结果:菌落PCR、酶切鉴定和测序表明重组质粒pLVX-AE gp160构建正确,RT-PCR、GFP荧光及流式细胞术结果均显示gp160基因能在细胞TC-1中稳定表达,免疫组化结果也表明小鼠体内接种的细胞可以稳定表达HIV Gp160蛋白。结论:建立了稳定表达HIV-1 CRF01_AE亚型Gp160蛋白的TC-1细胞及小鼠模型,为HIV-1 CRF01_AE亚型HIV疫苗的临床前研究提供了可靠的体外、体内免疫原性评价工具,为该疫苗的进一步开发奠定了坚实的实验基础。     

HIV-1 Diversity,Drug-Resistant Mutations,and Viral Evolution among High-Risk Individuals in Phase II HIV Vaccine Trial Sites in Southern China

HIV-1 prevalence in Guangxi, China, has been growing since 1996, when the first case was reported. Over half of HIV-1 positive patients in Guangxi Province were injecting drug users (IDUs), possibly because of the province’s location near drug-trafficking routes. Since a phase II HIV vaccine trial is ongoing there, a current characterization of the subtypes of HIV-1 among IDUs in Guangxi would provide critical information for future HIV vaccine trials, as well as further control and prevention of HIV-1 transmission. Thus, we conducted a molecular epidemiological investigation of HIV-1 samples from 2008–2010 among IDUs in multiple cities in Guangxi Province. Our results, based on the gag/pol fragment, indicated a very high proportion (78.47%) of HIV-1 CRF08_BC recombinants, some CRF01_AE (15.38%) recombinants, and a low proportion of CRF07_BC (6.15%) recombinants among the IDUs. The high proportion of CRF08 HIV-1 strains among recent IDUs matches the vaccine candidate constructs. However, future vaccine development should also incorporate CRF01-targeted vaccine candidates. Distinct Env sequence evolution patterns were observed for CRF08_BC and CRF01_AE, indicating that different local selection pressures have been exerted on these two HIV-1 subtypes. Unique drug-resistant mutations were also detected, and our data indicate that HIV treatment programs should consider pre-existing drug-resistant mutations.     

HIV-1 Genetic Characteristics and Transmitted Drug Resistance among Men Who Have Sex with Men in Kunming,China

Background

Yunnan has been severely affected by HIV/AIDS in China. Recently, the reported prevalence of HIV-1 among men who have sex with men (MSM) in Yunnan was high in China. To monitor dynamic HIV-1 epidemic among Yunnan MSM, HIV-1 genetic characteristics and transmitted drug resistance (TDR) were investigated.

Methods

Blood samples from 131 newly HIV-1 diagnosed MSM were continuously collected at fixed sites from January 2010 to December 2012 in Kunming City, Yunnan Province. Partial gag, pol and env genes were sequenced. Phylogenetic, evolutionary and genotypic drug resistance analyses were performed.

Results

Multiple genotypes were identified among MSM in Kunming, including CRF01_AE (64.9%), CRF07_BC (25.2%), unique recombinant forms (URFs, 5.3%), subtype B (3.1%) and CRF08_BC (1.5%). CRF01_AE and CRF07_BC were the predominant strains. The mean of genetic distance within CRF01_AE were larger than that within CRF07_BC. The estimated introducing time of CRF01_AE in Yunnan MSM (1996.9) is earlier than that of CRF07_BC (2002.8). In this study, subtype B was first identified in Yunnan MSM. CRF08_BC seems to be the distinctive strain in Yunnan MSM, which was seldom found among MSM outside Yunnan. The proportion of URFs increased, which further contributed to genetic diversity among MSM. Strikingly, genetic relatedness was found among these strains with MSM isolates from multiple provinces, which suggested that a nationwide transmission network may exist. TDR-associated mutations were identified in 4.6% individuals. The multivariate analysis revealed that non-native MSM and divorced/widowed MSM were independently associated with a higher TDR rate.

Conclusion

This work revealed diverse HIV-1 genetics, national transmission networks and a baseline level of TDR in MSM. These findings enhance our understanding of the distribution and evolution of HIV-1 in MSM, and are valuable for developing HIV prevention strategies for MSM.     

Susceptibility of HIV-1 subtypes B', CRF07_BC and CRF01_AE that are predominantly circulating in China to HIV-1 entry inhibitors

Background

The B′, CRF07_BC and CRF01_AE are the predominant HIV-1 subtypes in China. It is essential to determine their baseline susceptibility to HIV entry inhibitors before these drugs are used in China.

Methodology/Principal Findings

The baseline susceptibility of 14 representative HIV-1 isolates (5 CRF07_BC, 4 CRF01_AE, and 5 B′), most of which were R5 viruses, obtained from drug-naïve patients to HIV entry inhibitors, including two fusion inhibitors (enfuvirtide and C34), two CCR5 antagonists (maraviroc and TAK779) and one CXCR4 antagonist (AMD3100), were determined by virus inhibition assay. The sequences of their env genes were amplified and analyzed. These isolates possessed similar susceptibility to C34, but they exhibited different sensitivity to enfuvirtide, maraviroc or TAK779. CRF07_BC isolates, which carried polymorphisms of A578T and V583I in the N-terminal heptad repeat and E630Q, E662A, K665S, A667K and S668N in the C-terminal heptad repeat of gp41, were about 5-fold less sensitive than B′ and CRF01_AE isolates to enfuvirtide. Subtype B′ isolates with a unique polymorphism site of F317W in V3 loop, were about 4- to 5-fold more sensitive than CRF07_BC and CRF01_AE isolates to maraviroc and TAK779. AMD3100 at the concentration as high as 5 µM exhibited no significant inhibitory activity against any of the isolates tested.

Conclusion

Our results suggest that there are significant differences in baseline susceptibility to HIV entry inhibitors among the predominant HIV-1 subtypes in China and the differences may partly result from the naturally occurring polymorphisms in these subtypes. This study provides useful information for rational design of optimal therapeutic regimens for HIV-1-infected patients in China.     

After 18 months of antiretroviral therapy,total HIV DNA decreases more pronouncedly in patients infected by CRF01_AE than in those infected by subtype B and CRF07_BC

Whether the amount of HIV DNA is associated with the subtype of HIV‐1 after antiretroviral therapy (ART) has not been reported. In the present study, the amount of HIV DNA and RNA and CD4+T counts in blood and semen prior to and after 18 months of ART were compared in 48 patients infected by CRF01_AE, subtype B or CRF07_BC of HIV‐1. Viral RNA was suppressed and CD4 cell count recovery achieved in all patients. The level of HIV DNA were similar before ART; however, patients with CRF01_AE had less HIV DNA after ART than those with subtype B and CRF07_BC infection. According to prediction of co‐receptor usage by Geno2Pheno and PSSM in combination, more than 35.6% of clones for CRF01_AE were predicted as CXCR4‐using before ART, whereas less than 6% of those for subtype B and CRF07_BC were predicted as CXCR4‐using. After 18 months of ART, no CXCR4‐using clones were predicted in any of the subtypes. Despite more HIV RNA and fewer CD4 + T cells in patients with CRF01_AE before therapy, no significant differences (P > 0.05) in viral RNA or CD4 cell counts were observed between the subtypes after 18 months of ART. Thus, 18 months of antiretroviral therapy was more efficient in patients with CRF01_AE. Considering that successful ART dramatically reduces the viral load in both blood and semen, risks of sexual transmission of HIV were reduced, contributing to prevention of rapid spread of HIV among men who have sex with men in the region.

     

Characteristics of HIV-1 Molecular Epidemiology in Suzhou,China, from 2009 to 2014

正Dear Editor,Human immunodeficiency virus(HIV)and acquired immunodeficiency syndrome(AIDS)remain major public health problems in China,where approximately 780,000people currently live with HIV,according to the 2012 China AIDS Response Progress Report.Suzhou was an early epidemic region in Jiangsu province and the current situation of     

HIV-1 Genetic Diversity and Its Impact on Baseline CD4+T Cells and Viral Loads among Recently Infected Men Who Have Sex with Men in Shanghai,China

The HIV-1 epidemic among men who have sex with men (MSM) has been spreading throughout China. Shanghai, a central gathering place for MSM, is facing a continuously increasing incidence of HIV-1 infection. In order to better understand the dynamics of HIV-1 diversity and its influence on patient’s immune status at baseline on diagnosis, 1265 newly HIV-1-infected MSM collected from January 2009 to December 2013 in Shanghai were retrospectively analyzed for genetic subtyping, CD4+T cell counts, and viral loads. HIV-1 phylogenetic analysis revealed a broad viral diversity including CRF01_AE (62.13%), CRF07_BC (24.51%), subtype B (8.06%), CRF55_01B (3.24%), CER67_01B (0.95%), CRF68_01B (0.4%), CRF08_BC (0.08%) and CRF59_01B (0.08%). Twenty-four unique recombination forms (URFs) (1.98%) were identified as well. Bayesian inference analysis indicated that the introduction of CRF01_AE strain (1997) was earlier than CRF07_BC strain (2001) into MSM population in Shanghai based on the time of the most recent common ancestor (tMRCA). Three epidemic clusters and five sub-clusters were found in CRF01_AE. Significantly lower CD4+T cell count was found in individuals infected with CRF01_AE than in those infected with CRF07_BC infection (P<0.01), whereas viral load was significantly higher those infected with CRF01_AE than with CRF07_BC (P<0.01). In addition, the patients with >45 years of age were found to have lower CD4+T cell counts and higher viral loads than the patients with <25 years of age (P<0.05). This study reveals the presence of HIV-1 subtype diversity in Shanghai and its remarkable influence on clinical outcome. A real-time surveillance of HIV-1 viral diversity and phylodynamics of epidemic cluster, patient’s baseline CD4+T cell count and viral load would be of great value to monitoring of disease progression, intervention for transmission, improvement of antiretroviral therapy strategy and design of vaccines.     

A Comprehensive Mapping of HIV-1 Genotypes in Various Risk Groups and Regions across China Based on a Nationwide Molecular Epidemiologic Survey

Background

China is experiencing a dynamic HIV/AIDS epidemic. While serology based surveillance systems have reported the spread of HIV/AIDS, detailed tracking of its transmission in populations and regions is not possible without mapping it at the molecular level. We therefore conducted a nationwide molecular epidemiology survey across the country.

Methods

HIV-1 genotypes were determined from 1,408 HIV-positive persons newly diagnosed in 2006. The prevalence of each genotype was estimated by weighting the genotype’s prevalence from each province- and risk-specific subpopulation with the number of reported cases in the corresponding subgroups in that year.

Results

CRF07_BC (35.5%), CRF01_AE (27.6%), CRF08_BC (20.1%), and subtype B'' (9.6%) were the four main HIV-1 strains in China. CRF07_BC and CRF08_BC were the primary drivers of infection among injecting drug users in northeastern and southeastern China, respectively, and subtype B'' remained dominant among former plasma donors in central China. In contrast, all four strains occurred in significant proportions among heterosexuals nationwide, pointing to an expansion of the HIV-1 epidemic from high-risk populations into the general population. CRF01_AE also replaced subtype B as the principal driver of infection among men-who-have-sex-with-men.

Conclusions

Our study provides the first comprehensive baseline data on the diversity and characteristics of HIV/AIDS epidemic in China, reflecting unique region- and risk group-specific transmission dynamics. The results provide information critical for designing effective prevention measures against HIV transmission.     

Genotypes and Transmitted Drug Resistance among Treatment-Naive HIV-1-Infected Patients in a Northwestern Province,China: Trends from 2003 to 2013

Background

Transmitted drug resistance (TDR) reduces the efficacy of initial antiretroviral treatment and has become a public health concern. Little information is available regarding the genetic diversity of HIV-1 and the prevalence of TDR among treatment-naïve patients in a northwestern province of China since the implementation of national free antiretroviral therapy (ART).

Methods

Blood samples from 372 HIV-1 treatment-naive patients were collected between 2003 and 2013 in Shaanxi province. Viral RNA was extracted for nested PCR, and phylogenetic reconstruction and recombination analyses were performed to characterize patterns of the HIV-1 subtypes. Genotypic drug resistance testing was performed using an in-house assay to determine trends in the prevalence of HIV-1 transmitted drug resistance.

Results

Multiple genotypes were identified among the patients in Shaanxi, including B (25.0%), C (0.3%), G (0.3%), and CRF01_AE (39.2%), CRF07_BC (32.7%), CRF08_BC (0.8%), CRF55_01B (1.1%), and URFs (0.6%). The subtypes were associated with the transmission routes (χ² = 77.113, p<0.01). In this study, a low baseline CD4⁺ T cell count and a high viral load were found among CRF01_AE-infected patients compared with patients who were infected with non-CRF01_AE (p<0.01) through sexual transmission; however, the CRF01_AE subtype was not associated with a low baseline CD4⁺ T cell count or a high viral load in Chinese patients infected through blood transmission (p = 0.249). The overall TDR rate in this population was 4.4% between 2003 and 2013. A univariate logistic regression model revealed that a low CD4 T cell count (≤100 cells/µL) was associated with the development of drug-resistant strains.

Conclusion

Our work revealed diverse HIV-1 subtype distributions in Shaanxi province. We identified a low and stable TDR time trend among ART-naive patients. These findings enhance our understanding of HIV-1 genetic diversity and provide some guidelines for the improvement and implementation of a comprehensive public health strategy of HIV-1 TDR prevention.