Tissue and Cell Reservoirs for HIV

超出提供的空间。治疗艾滋病毒/艾滋病的成功关键取决于对隐匿性感染的彻底和准确的了解-在哪里以及如何保持艾滋病毒的持久性。这项建议侧重于将巨噬细胞和滤泡树突状细胞(FDC)作为HIV的细胞储存库，并将骨髓和脑作为组织储存库。尽管它们受到的关注相对较少，但这些细胞和组织隔间很可能代表着相当重要的艾滋病毒储存库。重要的是，每一家似乎都有能力成为潜在艾滋病毒感染的避难所。我们建议在这里确定这些细胞和组织是否真的可以作为潜伏感染的宿主，能够产生能够播种到身体的传染性颗粒。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传播模式，并确定大脑是否可以在外围重新接种艾滋病毒。我们还试图确定抗逆转录病毒药物敏感和耐药艾滋病毒的细胞和组织储存库。在这些研究中，将使用同时收集的死前骨髓和切除的颈淋巴活检样本，以及与患者匹配的血浆、脑脊液(CSF)和同时收集的分离的单核细胞和T淋巴细胞样本，以及在活检之前和之后进行HIV基因分型。将检查HIV基因组的两个区域，包膜基因的V1-V4区域，以及包括蛋白酶基因和逆转录酶基因的1.2kb片段，逆转录酶基因编码大多数重要的抗逆转录病毒耐药突变。为了更好地解决大脑能否在外周植入HIV的问题，我们将不仅评估存在于淋巴(LN)CD4+T细胞群体中的HIV物种，而且评估与LN生发中心滤泡树突状细胞(FDC)相关的HIV物种。这些调查将与对身体组织的研究相辅相成。此外，我们试图确定骨髓中与全身性病毒学衰竭相关的事件。将关注HIV在骨髓中的表达和单核/巨噬细胞的激活。将使用免疫组织化学和流式细胞术方法。这些研究将产生关于艾滋病毒宿主的重要新信息，以及骨髓事件对艾滋病毒免疫发病机制和疾病进展的贡献。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED. 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 excisional 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. PERFORMANCE SITE ========================================Section End===========================================