Monno Laura, Punzi Grazia, Scarabaggio Teresa, Saracino Annalisa, Brindicci Gaetano, Fiore José Ramon, Iambrenghi Onofrio Caputi, Di Stefano Mariantonietta, Pastore Giuseppe, Angarano Gioacchino
Clinic of Infectious Diseases, University of Bari, Bari, Italy.
J Med Virol. 2003 May;70(1):1-9. doi: 10.1002/jmv.10354.
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.
对接受高效抗逆转录病毒治疗（HAART）的1型人类免疫缺陷病毒（HIV-1）阳性患者的血液及同期直肠活检样本进行检测，通过对逆转录酶（RT）和蛋白酶（PR）区域进行直接测序来分析基因型耐药性，以比较这些区室中的耐药模式。研究了14名受试者（5名血浆病毒载量（pVL）不可检测，9名持续病毒血症）。5名pVL不可检测的患者中有4名黏膜HIV RNA也不可检测；前病毒DNA（外周血单核细胞和直肠活检）的序列分析未发现或仅发现少数耐药相关突变，且药敏谱无改变。所有病毒血症患者以及1名pVL阴性患者的黏膜HIV RNA水平均可检测到（1.93 - 4.21 log₁₀拷贝/毫克）；还获得了HIV RNA（血浆和/或直肠活检）的序列，检测到多个通常与当前/过去用药相符的突变。总体而言，共分析了40条HIV-1 PR序列和42条RT序列，产生了总共42对PR序列和47对RT序列（血浆/组织-RNA；血浆-RNA/组织-DNA；外周血单核细胞/组织-DNA；组织-DNA/RNA；组织-RNA/外周血单核细胞-DNA；外周血单核细胞-DNA/血浆-RNA），这些序列在总氨基酸水平上几乎总是存在差异（PR的中位不一致百分比为8.08%，RT为4.8%）。PR和RT序列对中耐药位点不一致的中位百分比分别为88.8%（四分位间距 = 20 - 100）和74.55%（四分位间距 = 31.75 - 100%）。通过计算机辅助解读突变谱检测到不同的耐药水平。结果支持HIV基因型在不同身体区室中的多样进化，并强调肠道黏膜在HIV基因型选择中的作用。应使用来自不同组织的样本进行耐药性评估，以全面了解接受抗逆转录病毒治疗患者的耐药情况。
