Marras Daniele, Bruggeman Leslie A, Gao Feng, Tanji Nozomu, Mansukhani Mahesh M, Cara Andrea, Ross Michael D, Gusella G Luca, Benson Gary, D'Agati Vivette D, Hahn Beatrice H, Klotman Mary E, Klotman Paul E
Division of Nephrology, Department of Medicien, Mount Sinai School of Medicine, New York, New York, USA.
Nat Med. 2002 May;8(5):522-6. doi: 10.1038/nm0502-522.
HIV-associated nephropathy is a clinicopathologic entity that includes proteinuria, focal segmental glomerulosclerosis often of the collapsing variant, and microcystic tubulointerstitial disease. Increasing evidence supports a role for HIV-1 infection of renal epithelium in the pathogenesis of HIV-associated nephropathy. Using in situ hybridization, we previously demonstrated HIV-1 gag and nef mRNA in renal epithelial cells of patients with HIV-associated nephropathy. Here, to investigate whether renal epithelial cells were productively infected by HIV-1, we examined renal tissue for the presence of HIV-1 DNA and mRNA by in situ hybridization and PCR, and we molecularly characterized the HIV-1 quasispecies in the renal compartment. Infected renal epithelial cells were removed by laser-capture microdissection from biopsies of two patients, DNA was extracted, and HIV-1 V3-loop or gp120-envelope sequences were amplified from individually dissected cells by nested PCR. Phylogenetic analysis of kidney-derived sequences as well as corresponding sequences from peripheral blood mononuclear cells of the same patients revealed evidence of tissue-specific viral evolution. In phylogenetic trees constructed from V3 and gp120 sequences, kidney-derived sequences formed tissue-specific subclusters within the radiation of blood mononuclear cell-derived viral sequences from both patients. These data, along with the detection of HIV-1-specific proviral DNA and mRNA in tubular epithelium cells, argue strongly for localized replication of HIV-1 in the kidney and the existence of a renal viral reservoir.
人类免疫缺陷病毒相关性肾病是一种临床病理实体，包括蛋白尿、常为塌陷型的局灶节段性肾小球硬化以及微囊性肾小管间质性疾病。越来越多的证据支持肾上皮细胞的HIV-1感染在人类免疫缺陷病毒相关性肾病发病机制中起作用。我们之前利用原位杂交技术在人类免疫缺陷病毒相关性肾病患者的肾上皮细胞中证实了HIV-1 gag和nef mRNA的存在。在此，为了研究肾上皮细胞是否被HIV-1有效感染，我们通过原位杂交和聚合酶链反应检测肾组织中HIV-1 DNA和mRNA的存在情况，并对肾组织中的HIV-1准种进行分子特征分析。通过激光捕获显微切割从两名患者的活检组织中去除受感染的肾上皮细胞，提取DNA，并通过巢式聚合酶链反应从单个切割的细胞中扩增HIV-1 V3环或gp120包膜序列。对肾脏来源序列以及同一患者外周血单个核细胞的相应序列进行系统发育分析，揭示了组织特异性病毒进化的证据。在由V3和gp120序列构建的系统发育树中，肾脏来源序列在两名患者血液单个核细胞来源的病毒序列辐射范围内形成了组织特异性亚群。这些数据，连同在肾小管上皮细胞中检测到HIV-1特异性前病毒DNA和mRNA，有力地证明了HIV-1在肾脏中的局部复制以及肾脏病毒库的存在。
