Antiretroviral-Drug Resistant Mutations and Genetic Diversity in HIV-1 Infected Individuals in Nigeria

Introduction: Despite the success derived from antiretroviral therapy, drug resistance (DR) mutations are known to develop and are major impediments to treatment of HIV patients. Therefore, periodic assessment of HIVDR is needed to ensure continuous HAART efficacy. This study assessed the magnitude of drug resistance as well as HIV genetic variability in drug-naive and treated patients in Nigeria. Methodology: Genotypic analysis was performed by sequencing plasma specimens from 40 individuals in a cross sectional study involving 202 HIV infected patients from all the six geopolitical zones of Nigeria. Sequences were analyzed for presence of HIVDR mutation using the algorithm in Stanford HIVDR database and confirmed by IAS-USA 2009 mutation list. Phylogenetic and recombination analyses were done using PAUP V4.0 and REGA V2.0 respectively. Results: Major DR mutations were detected in the reverse transcriptase (RT) gene of 5 (33%) drug experienced and 2 (8%) na?ve patients. Most common mutations were M184V and K103N with no protease (PR) mutations detected. Thymidine analogue mutations (TAMs) and a complex multi resistance mutation Q151M were detected in 3 samples. Polymorphic substitutions were observed in both PR and RT gene. Phylogenetic analysis revealed Group M isolates of G (20), J (1), circulating recombinant forms: CRF02_AG (14), CRF-18-cpx (1), CRF06_cpx (3) and a unique AD recombinant (1). Conclusion: Our findings corroborate previous studies on circulating DR viruses in Nigeria while genetic diversity is on the increase. In view of ART scale-up, monitoring the resistance pattern and genetic diversity will aid in appropriate prevention strategies.

Exploration of an Efficient Simultaneous Molecular Detection Method of HIV,HCV,and Syphilis from a Single Dried Blood Spot

Objective The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA,HIV-1 DNA,and HCV RNA using one dried blood spot(DBS)as an alternative sample to plasma.Method A total of 571 paired DBS/plasma samples were collected from men who have sex with men(MSM)and injection drug users(IDUs),and serological and molecular assays were performed.Using plasma results as the reference standard,the performance of DBS tests for HIV-1 RNA,HIV-1 DNA,and HCV RNA was evaluated.Pearson’s correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma.Results Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA,five samples(5/32)were not detectable in DBS,while measurable HIV-1 RNA levels were present in plasma(1.44 to3.99 log10 copies/m L).There were two samples(2/94)with undetectable HCV RNA in DBS,while measurable HCV RNA levels were present in plasma(-5 to 5.99 log10 copies/m L).The correlation between HIV-1 RNA light chain variable region(VL)values obtained from plasma and DBS showed that r=0.683(P<0.01),n=27 and r=0.612(P<0.01),n=89 in HCV RNA.Bland-Altman analysis revealed that in HIV-1 RNA,the mean(±SD)difference between HIV-1 RNA in plasma and DBS was 1.00±1.01 log10 copies/m L,and all samples were within±1.96 SD(-0.97 to 2.97 log10 copies/m L)for DBS.The mean difference(±SD)in HCV RNA was 0.15±1.08 log10 copies/m L,and 94.38%(84/89)were within±1.96 SD(-1.96 to 2.67 log10 copies/m L).Overall,HIV-1 RNA and HCV RNA levels obtained from a DBS were lower than those obtained from plasma.HIV-1 DNA in a DBS showed concordant results with HIV-1 RNA in plasma.HIV-1 DNA RT-PCR using a DBS showed acceptable performance.Conclusion The performance of the simultaneous detection of HIV-1 RNA,HIV-1 DNA,and HCV RNA using one DBS was acceptable.DBS,as an alternative sample to plasma,may be a viable option for the simultaneous detection of HIV-1 RNA,HIV-1 DNA,and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.

The Pattern of Haematological Changes in the Baseline Blood Cell Counts and the CD4⁺T Lymphocyte Levels among Antiretroviral Therapy Naïve Adult HIV Positive Patients in a Nigerian Hospital

Background: Haematological abnormalities are strong independent predictors of morbidity and mortality in HIV infection. Objectives of the Study: This study was carried out to assess the pattern of the changes in the baseline peripheral blood cell counts among adult HIV positive patients. To also determine the CD4+ T cell levels and its correlation with the changes in the baseline cell counts of the patients using HIV negative blood donors as controls. Methods: This was a case controlled prospective study. The subjects were antiretroviral therapy naive adult HIV positive patients and HIV negative blood donor controls. Five milliliters (5 mls) of venous blood was collected from the cubitus of every consecutive consenting subject. Blood sample was analysed for baseline complete blood cell counts and the CD4+ T lymphocyte levels using Sysmex and Cyflow R Counter autoanalysers respectively. Obtained data was analysed with the statistical package for the social scientist (SPSS version 20.0). The Erythrocyte sedimentation rate of subjects was measured by the manual standard Westergreen method. Results: Of 300 subjects, there were 139 (46.3%) males and 161 (53.7%) females in the study. Anaemia was found in 72 (29.2%), leucopenia in 20 (8%) and thrombocytopenia in 6 (2.4%) of the patients. The mean erythrocyte sedimentation rates of the subjects were 81.88 mm/hr and 9.46 mm/hr (p = 0.000) for the patients and the controls respectively. The mean CD4+ T lymphocyte cell counts were 293 cells/μl and 750 cells/μl (p = 0.000) for the patients and controls respectively. Conclusion: Anaemia, leucopenia and thrombocytopenia were more prevalent among the studied HIV positive patients.

我国HIV感染者CD_4^+T淋巴细胞检测能力的研究

目前，人类关于HIV感染的检测项目很多，其中CD4^＋ T淋巴细胞的检测已成为评估HIV感染者疾病状况切实可行和相对经济的检测项目，CD4^＋ T淋巴细胞水平用来监测不同临床阶段HIV感染者免疫状况的变化，判定HIV感染者病程，预测机会性感染的出现以及评价抗HIV药物疗效。从2006年起准确检测CD4^＋ T淋巴细胞计数已经成为判断是否开始治疗AIDS和HIV感染者以及治疗效果观察对比的关键。国际上如美国CDC、英国国家室间质量评价系统（The United Kingdom National External Quality Assessment Service，NEQAS）、加拿大HIV／AIDS质量评价和免疫标准化体系（Quality Assessment and Standardization for Immunological Measures Relevant to HIV／AIDS，QASI）定期发放质控品，

Preferential loss of gut-homing a4β7 CD4＋T cells and their circulating functional subsets in acute HIV-1 infection

Preferential infection and depletion of gut-homing a4β7 CD4＋ T cells in the blood are observed in chronic HIV/SIV infection. The dynamic change in gut-homing a4p7 CD4＋ T cells and their functional subsets during the acute stages of HIV-1 infection are less documented. Therefore, we conducted a cohort study to investigate whether acute HIV-1 infection induced abnormalities in gut-homing a4β7 CD4＋ T cells and their functional subsets. We examined the frequency, absolute number, and functionality of gut-homing a4β7 CD4＋ T cells in 26 acute HIV-l-infected patients compared with 20 healthy individuals. We found that circulating gut-homing a4β7 CD4＋ T cells were preferentially depleted during acute HIV-1 infection and were positively correlated with absolute CD4＋ T-cell count in blood. Notably, Th17 and Thl cell subsets of gut-homing CD4＋ T cells were also decreased, which resulted in an imbalance of T helper cells （Th 1）-regulatory T cells （Treg） and Treg.Th 17 ratios. Gut-homing Th17 and Thl cells were also positively correlated with the absolute number of total CD4＋ T cells and gut-homing CD4＋ T cells. The gut-homing Treg：Th17 ratio was inversely correlated with the CD4＋ T-cell count. Taken together, the analyses of our acute HIV-1 cohort demonstrate that gut-homing a4β7 CD4＋ T cells and their functional subsets were profoundly depleted during acute HIV-1 infection, which may have resulted in the persistent loss of circulating CD4＋ T cells and an imbalance of Thl-Treg and Treg.Th17 ratios and contribute to HIV-1 disease pathogenesis.