Tissue and Cell Reservoirs for HIV

描述（由申请人提供）：成功治疗艾滋病毒/艾滋病关键取决于对隐性感染的彻底和准确理解，即艾滋病毒在何处以及如何持续存在。这一建议的重点是巨噬细胞和滤泡树突状细胞（FDC）作为HIV的细胞储存库，骨髓和大脑作为组织储存库。尽管它们受到的关注相对较少，但这些细胞和组织隔室很可能代表了相当重要的HIV储存库。重要的是，每一种病毒似乎都能成为潜伏HIV感染的避难所。我们在此提议确定这些细胞和组织是否确实可以作为潜伏感染的宿主，能够产生可以在身体中播种的感染性颗粒。我们试图确定艾滋病毒在淋巴组织和中枢神经系统之间的运输模式，并确定大脑是否可以将艾滋病毒重新植入周围。我们还寻求鉴定抗逆转录病毒药物敏感和耐药HIV的细胞和组织储存库。在这些研究中，HIV基因分型将使用同时收集的死前骨髓和切除的颈部淋巴结活检标本，以及同时收集的患者匹配的血浆、脑脊液（CSF）和分离的血液单核细胞和t淋巴细胞标本，以及活检之前和之后的标本。HIV基因组的两个区域将被检查，包膜基因的V1-V4区域，以及一个1.2kb的片段，其中包括蛋白酶基因和逆转录酶基因的一部分，逆转录酶基因编码了大多数重要的抗逆转录病毒抗性突变。为了最佳地解决大脑是否可以将HIV植入外周的问题，我们将评估不仅存在于淋巴结（LN） CD4+ t细胞群中的HIV物种，还存在于LN生发中心滤泡树突状细胞（FDC）相关的HIV物种。这些调查将与死后组织研究相辅相成。此外，我们试图确定与系统病毒学失败相关的骨髓事件。我们将关注HIV在骨髓中的表达和单核/巨噬细胞的活化。将采用免疫组织化学和流式细胞术方法。这些研究将产生关于HIV储存库的重要新信息，以及骨髓事件对HIV免疫发病机制和疾病进展的贡献。

DESCRIPTION (provided by applicant): Success in treating HIV/AIDS is critically dependent upon a thorough and accurate understanding of occult infection---where and how HIV persistence is maintained. This proposal focuses on the macrophage and follicular dendritic cell (FDC) as cellular reservoirs for HIV, and bone marrow and brain as tissue reservoirs. Although they have received comparatively little attention, these cell and tissue compartments most probably represent HIV reservoirs of considerable significance. Importantly, each appears capable of serving as a sanctuary for latent HIV infection. We propose here to determine if, indeed, these cells and tissues can serve as latently infected hosts able to yield infectious particles that can seed the body. We seek to identify patterns of HIV trafficking between lymphoid tissues and the Central Nervous System, and determine if the brain can reseed the periphery with HIV. We also seek to identify cell and tissue reservoirs for antiretroviral drug-sensitive, versus drug-resistant, HIV. For these studies, HIV genotyping will be performed using contemporaneously-collected ante-mortem bone marrow and excised cervical lymph node biopsy specimens, in conjunction with patient-matched plasma, cerebrospinal fluid (CSF), and separated blood monocyte and T-lymphocyte specimens collected concomitantly, as well as prior to, and following biopsy. Two regions of the HIV genome will be examined, the V1-V4 region of the envelope gene, and a 1.2kb fragment which includes the protease gene and that portion of the reverse transcriptase gene which codes for most of the antiretroviral resistance mutations of significance. To optimally address the question of whether the brain can seed the periphery with HIV, we will evaluate the HIV species present not only within the lymph node (LN) CD4+ T-cell population, but also those associated with LN germinal center follicular dendritic cells (FDC). These investigations will be complemented with studies of post-mortem tissues. In addition, we seek to identify events in bone marrow associated with systemic virologic failure. Attention will be paid to HIV expression and monocyte/macrophage activation in marrow. Immunohistochemistry and flow cytometry methods will be used. These studies will yield important new information regarding HIV reservoirs, as well as the contributions of events in bone marrow to HIV immunopathogenesis and disease progression.