Trillo-Pazos G, Diamanturos A, Rislove L, Menza T, Chao W, Belem P, Sadiq S, Morgello S, Sharer L, Volsky D J
Division of Molecular Virology, St. Luke's-Roosevelt Hospital Center, College of Physicians & Surgeons, Columbia University, New York, NY 10019, USA.
Brain Pathol. 2003 Apr;13(2):144-54. doi: 10.1111/j.1750-3639.2003.tb00014.x.
In HIV-1 encephalitis, HIV-1 replicates predominantly in macrophages and microglia. Astrocytes also carry HIV-1, but the infection of oligodendrocytes and neurons is debated. In this study we examined the presence of HIV-1 DNA in different brain cell types in 6 paraffin embedded, archival post-mortem pediatric and adult brain tissues with HIV-1 encephalitis by Laser Capture Microdissection (LCM). Sections from frontal cortex and basal ganglia were stained by immunohistochemistry for CD68 (microglia), GFAP (astrocytes), MAP2 (neurons), and p24 (HIV-1 positive cells) and different cell types were microdissected by LCM. Individual cells or pools of same type of cells were lysed, the cell lysates were subjected to PCR using HIV-1 gag SK38/SK39 primers, and presence of HIV-1 DNA was confirmed by Southern blotting. HIV-1 gag DNA was consistently detected by this procedure in the frontal cortex and basal ganglia in 1 to 20 p24 HIV-1 capsid positive cells, and in pools of 50 to 100 microglia/macrophage cells, 100 to 200 astrocytes, and 100 to 200 neurons in HIV-1 positive cases but not in HIV-1 negative controls. These findings suggest that in addition to microglia, the infection of astrocytes and neurons by HIV-1 may contribute to the development of HIV-1 disease in the brain.
在HIV-1脑炎中，HIV-1主要在巨噬细胞和小胶质细胞中复制。星形胶质细胞也携带HIV-1，但少突胶质细胞和神经元的感染情况仍存在争议。在本研究中，我们通过激光捕获显微切割（LCM）技术，检测了6例石蜡包埋的、存档的HIV-1脑炎患儿和成人尸检脑组织中不同脑细胞类型中HIV-1 DNA的存在情况。额叶皮质和基底神经节的切片通过免疫组织化学染色检测CD68（小胶质细胞）、GFAP（星形胶质细胞）、MAP2（神经元）和p24（HIV-1阳性细胞），并通过LCM对不同细胞类型进行显微切割。单个细胞或同类型细胞池被裂解，细胞裂解物使用HIV-1 gag SK38/SK39引物进行PCR，通过Southern印迹法确认HIV-1 DNA的存在。通过该方法，在HIV-1阳性病例的额叶皮质和基底神经节中，在1至20个p24 HIV-1衣壳阳性细胞以及50至100个小胶质细胞/巨噬细胞、100至200个星形胶质细胞和100至200个神经元的细胞池中始终检测到HIV-1 gag DNA，但在HIV-1阴性对照中未检测到。这些发现表明，除了小胶质细胞外，HIV-1对星形胶质细胞和神经元的感染可能在脑部HIV-1疾病的发展中起作用。
