VIRAL LOAD IN CELLULAR SUBCOMPARTMENTS FOLLOWING HAART

与人类免疫缺陷病毒(HIV)新制剂的联合治疗极大地减缓了艾滋病和死亡的进展。这些治疗方法能够将外周血液中的病毒降低到检测到的水平以下。然而，病毒复制的储备库仍然存在，主要是在淋巴组织中，这可能会重新点燃艾滋病毒的复制。该项目检测了患者在高活性治疗前和治疗期间从切除的颈淋巴结组织和外周血中采集的一系列样本。它将建立一种机制，从几个隔间同时和可重复地获取样本。最先进的流式细胞术和细胞分选、基于等温核酸序列的分析(NASBA)HIV定量和非放射性同位素原位杂交技术将被用于检测这些样本中的细胞亚型(淋巴细胞与单核/巨噬细胞)HIV病毒载量。将研究细胞亚室随时间和隔室之间的变化模式，以试图了解治疗失败并为设计更有效的治疗策略提供新的见解。属于独特的应答创造的不和谐应答的患者(15%-20%的患者在治疗后经历病毒载量和CD4计数的上升，但似乎获得了与预期的病毒学应答相同的临床益处)，也将进行研究，以确定超出吸收、坚持和表型/基因耐药模式的治疗应答的病理生理学参数，作为病毒学应答的解释。这个项目将调查病毒嗜性的作用和治疗方案在特定淋巴细胞亚型中的特定活性，包括终末分化细胞和分裂细胞。对感染细胞子集的进一步表征代表了该项目的未来方向，即通过细胞周期蛋白A、B和D分析细胞周期，并将为特定治疗方案如何影响总病毒载量提供独特的见解。然而，这个研究生涯奖的最高目标是为PI提供教学培训，重叠密切监督的临床研究期间，从而使PI成为一个可行的独立研究人员。

Combination therapy with new agents against the human immunodeficiency virus (HIV) has dramatically slowed the progression to AIDS and death. These treatments are capable of reducing virus in the peripheral blood to below the levels of detection. However, reservoirs of viral replication persist, largely in lymph node tissue, that potentially reignite HIV replication. This project examines serial samples from excisional cervical lymph node tissue and peripheral blood in patients before and during highly active therapy. It will establish a mechanism for obtaining samples from several compartments simultaneously and reproducibly. The combination of state-of-the-art flow cytometry and cell sorting, isothermal Nucleic Acid Sequence Based Assay (NASBA) HIV quantitation, and a non-radioisotope in situ hybridization technique will be used to measure cellular subtype (lymphocyte versus monocyte/macrophage) HIV viral load in these samples. Patterns of changes in cellular subcompartments across time and between the compartments will be examined in an attempt to understand treatment failure and to provide new insights into the design of more effective treatment strategies. Patients who fall into the unique category of response - coined Discordant Responders (the 15 - 20 percent of patients who experience a rise in both viral load and CD4 count in response to therapy yet appear to reap the same clinical benefits as patients with the expected virologic response), will also be studied in order to determine pathophysiologic parameters of response to therapy that go beyond absorption, adherence, and phenotypic/genotypic resistance patterns as explanations for virologic response. This project will investigate the role of viral tropism and the specific activity of treatment regimens in specific lymphoid cell subtypes, including terminally differentiated versus dividing cells. A further characterization of the subset of infected cells represents the future direction of this project, namely analysis of cell cycle by cyclins A, B and D, and will provide unique insights in how specific treatment regimens might affect total viral load. However, the paramount goal of this research career award is to provide the PI with didactic training overlapping a closely supervised period of clinical investigation leading to establishment of the PI as a viable independent researcher.