Mutational patterns of paired blood and rectal biopsies in HIV‐infected patients on HAART

Abstract

Blood and concurrent rectal biopsy samples of human immunodeficiency virus type 1 (HIV-1)-positive highly active antiretroviral therapy (HAART)-treated patients were tested for genotypic resistance by direct sequencing of reverse transcriptase (RT) and protease (PR) regions to compare the patterns of resistance in these compartments. Fourteen subjects (five with undetectable plasma viral load (pVL) and nine persistently viremic) were studied. Four of five patients with undetectable pVL also had undetectable mucosal HIV RNA; sequence analyses from proviral DNA (PBMCs and rectal biopsy) were obtained with none or few resistance-associated mutations and no alteration of susceptibility profile. All viremic patients, and one with negative pVL, had detectable levels of mucosal HIV RNA (1.93-4.21 log(10) copies/mg); sequences of HIV RNA (plasma and/or rectal biopsy) were also obtained, and multiple mutations generally compatible with current/past medications were detected. Overall, 40 HIV-1 PR and 42 RT sequences were analyzed, yielding a total of 42 PR and 47 RT sequence pairs (plasma/tissue-RNA; plasma-RNA/tissue-DNA; PBMC/tissue-DNA; tissue-DNA/RNA; tissue-RNA/PBMC-DNA; PBMC-DNA/plasma-RNA), which almost always differed at the total amino acid level (median percentage discordance 8.08% in the PR, 4.8% in RT). The median percentage of resistance position discordance equaled 88.8% (IQR = 20-100) in the PR and 74.55% (IQR = 31.75-100%) in the RT pairs, respectively. Different resistance levels were detected by means of a computer-assisted interpretation of mutational profiles. The results support the multiform evolution of HIV genotype in various body compartments and emphasize the participation of intestinal mucosa in HIV genotype selection. Samples from diverse tissues should be used for resistance evaluation to obtain a complete picture of drug resistance for antiretroviral-treated patients.

Aspectos funcionais, microbiológicos e morfológicos intestinais em crianças infectadas pelo vírus da imunodeficiência humana

“…CLAYTON et al (8) documentaram progressão qualitativa e quantitativa nas alterações vistas na mucosa retal de indivíduos infectados pelo HIV, a qual se correlacionou com a presença e a quantidade do vírus no tecido colônico, sugerindo, além da enteropatia do HIV anteriormente comentada, que a mucosa intestinal poderia constituir local de proliferação e de destruição de linfócitos. Mais recentemente, MONNO et al (21) , comparando amostras de HIV obtidas concomitantemente do sangue e de tecido retal, verifi caram padrões de resistência aos anti-retrovirais diferentes, dando suporte à idéia de evolução genotípica variável do HIV nos diferentes compartimentos corporais e enfatizando a participação da mucosa intestinal na seleção do genótipo do HIV.…”

The Genotyping Resistance Profile of the <b><i>Pol</i></b> Gene Detected in Blood of Newly Diagnosed HIV-Positive Men Is Durably Archived in the Gut Whatever the Time of Initiation of cART

“…The time to obtain HIV RNA undetectability in blood plasma, defined as the period between initiation of cART and the first viral load value <40 copies/mL, was 4 months [2,3,4,5,6,7,8,9,10,11,12,13,14,15]. No viral rebound was observed during the study period except 1 blip in patient 4 at month 18 after starting cART.…”

“…The rectal mucosa can integrate HIV-1 strains resistant to antiretroviral treatment selected during therapy and participates to the failure to reach optimal local drug concentrations [7,9,10]. Josefsson et al [3] showed that HIV-1 DNA integrated in GALT-infected T cells remains stable during suppressive cART when sequences between plasma sampled at the time of initiation of cART and GALT biopsies obtained 8-12 years after starting therapy were compared.…”

“…In addition, differences in the resistance profiles have been observed between anatomical sites including the gut, suggesting the participation of intestinal mucosa in HIV genotype selection [7,9]. It has also been observed that successive drug-resistant variants selected during nonsuppressive therapy remain archived in the cellular reservoir after long-term suppressive cART [10].…”

HIV infection and protease genetic diversity in a rural area of the Southern Central African Republic

“…At testing, 21% of these anti-HIV-positive women had syphilis, 18.4% gonorrhoea, and an additional 5.2% gardnerella vaginosis. DNA was extracted from the HIV-positive bloodspots (QIAamp DNA blood Mini kit, Quiagen, Germany), and amplification of the pol gene was performed using an ''in-house'' protocol [Monno et al, 2003]. Thereafter, the PCR products were directly sequenced (BigDye Terminator Cycle Sequencing Kit, Applied Biosystem, Foster City, CA).…”