HIV-1 pol Diversity among Female Bar and Hotel Workers in Northern Tanzania

A national ART program was launched in Tanzania in October 2004. Due to the existence of multiple HIV-1 subtypes and recombinant viruses co-circulating in Tanzania, it is important to monitor rates of drug resistance. The present study determined the prevalence of HIV-1 drug resistance mutations among ART-naive female bar and hotel workers, a high-risk population for HIV-1 infection in Moshi, Tanzania. A partial HIV-1 pol gene was analyzed by single-genome amplification and sequencing in 45 subjects (622 pol sequences total; median number of sequences per subject, 13; IQR 5–20) in samples collected in 2005. The prevalence of HIV-1 subtypes A1, C, and D, and inter-subtype recombinant viruses, was 36%, 29%, 9% and 27%, respectively. Thirteen different recombination patterns included D/A1/D, C/A1, A1/C/A1, A1/U/A1, C/U/A1, C/A1, U/D/U, D/A1/D, A1/C, A1/C, A2/C/A2, CRF10_CD/C/CRF10_CD and CRF35_AD/A1/CRF35_AD. CRF35_AD was identified in Tanzania for the first time. All recombinant viruses in this study were unique, suggesting ongoing recombination processes among circulating HIV-1 variants. The prevalence of multiple infections in this population was 16% (n = 7). Primary HIV-1 drug resistance mutations to RT inhibitors were identified in three (7%) subjects (K65R plus Y181C; N60D; and V106M). In some subjects, polymorphisms were observed at the RT positions 41, 69, 75, 98, 101, 179, 190, and 215. Secondary mutations associated with NNRTIs were observed at the RT positions 90 (7%) and 138 (6%). In the protease gene, three subjects (7%) had M46I/L mutations. All subjects in this study had HIV-1 subtype-specific natural polymorphisms at positions 36, 69, 89 and 93 that are associated with drug resistance in HIV-1 subtype B. These results suggested that HIV-1 drug resistance mutations and natural polymorphisms existed in this population before the initiation of the national ART program. With increasing use of ARV, these results highlight the importance of drug resistance monitoring in Tanzania

Identification of CRF10_CD viruses among bar and hotel workers in Moshi, Northern Tanzania.

We recently identified an HIV-1 subtype C and D circulating recombinant form (CRF10_CD) in infants in Dar es Salaam, Tanzania. So far, this is the only reported HIV-1 CRF in East Africa. However, evidence for its spread in the adult population is scarce. Here we describe the presence of CRF10_CD in two asymptomatic bar and hotel workers in Moshi, Northern Tanzania. Subgenomic sequences from gag (3'p24-5'p7), env (C2-C5), and the 5' LTR were used for phylogenetic analysis and identification of recombination. Genetic divergence between the CRF10_CD sequences from Moshi suggested that they were contracted from independent sources. A third bar worker was infected with an apparent CRF10_CD/subtype A recombinant virus. Our data indicate that CRF10_CD genomes can be transmitted both vertically and heterosexually

HIV type 1 subtypes among bar and hotel workers in Moshi, Tanzania.

The HIV-1 prevalence among bar and hotel workers in Tanzania suggests they are a high-risk group for HIV-1 infection. We determined the HIV-1 subtype of 3'-p24/5'-p7 gag and C2-C5 env sequences from 40 individuals representing this population in Moshi. Genetic patterns composed of A(gag)-A(env), C(gag)-C(env), and D(gag)-D(env) were found in 19 (48.0%), 8 (20.0%), and 3 (8.0%) samples, respectively. The remaining 10 samples (25%) had different subtypes in gag and env, indicative of intersubtype recombinants. Among these recombinants, two contained sequences from HIV-1 subsubtype A2, a new genetic variant in Tanzania. Five bar and hotel workers may have been infected with viruses from a common source, based on phylogenetic analysis. The information obtained by surveillance of HIV-1 subtypes in a high-risk population should be useful in the design and evaluation of behavioral, therapeutic, and vaccine trial interventions aimed at reducing HIV-1 transmission

Patterns of acquired HIV-1 drug resistance mutations and predictors of virological failure in Moshi, Northern Tanzania.

Emergence of HIV drug resistance poses a serious risk of inactivity to all currently approved antiretroviral drugs. Profiles of HIV drug resistance mutations (HIVDRM) and virological failure (VF) are not extensively studied in Tanzania. This study aimed to determine HIVDRM and predictors of VF in HIV-infected individuals failing first-line HIV drugs in Moshi, Northern Tanzania. A case-control study was conducted at Kilimanjaro Christian Medical Centre, Mawenzi, Pasua and Majengo health facilities with HIV-care and treatment clinics from October, 2017 to August, 2018. Cases and controls were HIV-infected individuals with VF and viral suppression (VS) respectively. HIV-1 reverse transcriptase and protease genes were amplified and sequenced. Stanford University's HIV drug resistance database and REGA subtyping tool 3.0 determined HIVDRM and HIV-1 subtypes respectively. Odds ratios (OR) with 95% confidence interval (95% CI) investigated predictors of VF. P-value < 5% was considered statistically significant. A total of 124 participants were recruited, of whom 63 (50.8%) had VF, 61 (49.2%) had VS and 82 (66.1%) were females. Median [IQR] age and duration on ART were 45 [35-52] years and 72 [48-104] months respectively. Twenty-five out of 26 selected samples from cases were successfully sequenced. Twenty-four samples (96%) had at least one major mutation conferring resistance to HIV drugs, with non-nucleoside analogue reverse transcriptase inhibitor (NNRTI)-resistance associated mutations as the majority (92%). Frequent NNRTI-resistance associated mutations were K103N (n = 11), V106M (n = 5) and G190A (n = 5). Prevalent nucleoside analogue reverse transcriptase inhibitors-resistance associated mutations were M184V (n = 17), K70R (n = 7) and D67N (n = 6). Dual-class resistance was observed in 16 (64%) samples. Thirteen samples (52%) had at least one thymidine analogue-resistance associated mutation (TAM). Three samples (12%) had T69D mutation with at least 1 TAM. Two samples (8%) had at least one mutation associated with protease inhibitor resistance. Age [aOR = 0.94, 95% CI (0.90-0.97), p < 0.001] and occupation [aOR = 0.35, 95% CI (0.12-1.04), p = 0.059] associated with VF. In conclusion, HIV drug resistance is common among people failing antiretroviral therapy. Resistance testing will help to guide switching of HIV drugs

Herpes simplex virus type 2 infection among bar and hotel workers in northern Tanzania: prevalence and risk factors.

BACKGROUND: Type-specific serological tests have allowed for a better understanding of the epidemiology of herpes simplex virus type 2 (HSV-2) infection in Africa. GOAL: The goal was to determine risk factors for HSV-2 among bar and hotel workers in Moshi, Tanzania. STUDY DESIGN: A cross-sectional study was conducted involving 515 workers in randomly selected bars and hotels in Moshi. RESULTS: The seroprevalence of HSV-2 was 43.5%. Women were more likely to be HSV-2-seropositive than men (age-adjusted OR = 3.8; 95% CI = 2.5-5.8). In multivariate analyses, age was positively associated with HSV-2 in both women and men. HIV-1-seropositive women had a significantly increased risk of HSV-2 infection (adjusted OR = 2.8; 95% CI = 1.5-5.1). Other predictors of HSV-2 were religion and sexual behavior for women and level of education, frequency of alcohol use, and concurrent partners for men. CONCLUSION: The most common genital infection was that with HSV-2. Control of HSV-2 might be an important strategy for HIV-1 infection prevention in this population

The role of herpes simplex virus type 2 and other genital infections in the acquisition of HIV-1 among high-risk women in northern Tanzania.

BACKGROUND: We examined the role of herpes simplex virus type 2 (HSV-2) and other genital infections on human immunodeficiency virus type 1 (HIV-1) incidence in a cohort study conducted between 2002 and 2005 among female bar/hotel workers in Moshi, Tanzania. METHODS: At baseline and every 3 months thereafter, participants were interviewed, and blood and genital samples were collected. Predictors of HIV-1 incidence were evaluated using a Cox proportional hazards regression model. RESULTS: Of 845 women who were HIV-1 seronegative at baseline, 689 (81.5%) were monitored in the study for a total of 698.6 person-years at risk (PYARs). The overall HIV-1 incidence was 4.6/100 PYARs (95% confidence interval [CI], 3.0-6.2/100 PYARs), and condom use was very low. After adjustment for other risk factors, the risk of HIV-1 was increased among women with HSV-2 at baseline (hazard ratio [HR], 4.3 [95% CI, 1.5-12.4]) and in those who acquired HSV-2 during the study period (HR, 5.5 [95% CI, 1.2-25.4]). Other independent predictors of HIV-1 were baseline chlamydial infection (HR, 5.2), bacterial vaginosis (HR, 2.1), and the occurrence of genital ulcers (HR, 2.7). CONCLUSION: HSV-2 and other genital infections were the most important risk factors for HIV-1. Control of these infections could help to reduce HIV-1 incidence in this population

HIV-1 intra-subtype recombinant viruses among female bar and hotel workers in Moshi, Kilimanjaro region, Tanzania, during 2004–2007.

<p>The percentages of subjects with HIV-1 intra-subtype recombinants were calculated from analysis of the combined sets (early and later time points) of the viral quasispecies.</p

Maximum likelihood phylogenetic trees for three fragments of the one out of 57 viral quasispecies for subject 510 as identified by SimPlot analysis.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047605#pone-0047605-g005" target="_blank">Fig 5A</a>: A single viral sequence in subject 510 with distinct recombination pattern, D/U/D. The bootscan plot was generated by Simplot. HIV-1 subtype D is shown in the blue bar and unclassified region is shown in the black bar. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047605#pone-0047605-g005" target="_blank">Figs. 5B, 5C & 5D</a>: Maximum likelihood (ML) trees of three fragments classified as subtype D (nucleotide position 6,615 to 7,345, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047605#pone-0047605-g005" target="_blank">Fig. 5B</a>), unclassified fragment (nucleotide position 7,346 to 7,500, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047605#pone-0047605-g005" target="_blank">Fig. 5C</a>) and fragment classified as subtype D (nucleotide position 7,501 to 7,745). The viral quasispecies were analyzed with reference sequences from the Los Alamos HIV Sequence Database. The first fragment clustered with subtype D with a high aLRT support value (shown in light blue square), the second fragment clustered with subtype A1 but aLRT was insufficient (0.81; shown in black square). The third fragment clustered with subtype D with a weak aLRT support (0.87). Scale at the bottom of the Figure corresponds to 0.1 nucleotide substitutions per site.</p

HIV-1 single and multiple variants of the same subtype among female bar and hotel workers in Moshi, Kilimanjaro region, Tanzania.

*<p>A subset of 45 samples with pure subtypes (A1, C or D) was analyzed for multiple infections; sample 181 was excluded due to a single available DNA sequence.</p