VIRAL LOAD IN CELLULAR SUBCOMPARTMENTS FOLLOWING HAART

HAART 后细胞亚区室中的病毒载量

• 批准号: 2898530
• 负责人: DAVID M ASMUTH
• 金额: $ 14万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2898530
• 关键词: AIDS therapy; HIV infections; antiAIDS agent; cell cycle; clinical research; cyclins; human immunodeficiency virus; human subject; hydroxyurea; immunocytochemistry; in situ hybridization; lymph nodes; monocyte; virus classification; virus load; virus replication

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.

对抗人类免疫缺陷病毒（HIV）的新药物的联合治疗极大地减缓了艾滋病的进展和死亡。 这些治疗能够将外周血液中的病毒减少到检测水平以下。 然而，病毒复制库仍然存在，主要存在于淋巴结组织中，这可能会重新引发艾滋病毒复制。 该项目在高度积极的治疗之前和期间检查了患者切除的颈部淋巴结组织和外周血的系列样本。 它将建立一种同时、可重复地从多个隔间获取样本的机制。 最先进的流式细胞术和细胞分选、等温核酸序列分析 (NASBA) HIV 定量以及非放射性同位素原位杂交技术的结合将用于测量这些样本中的细胞亚型（淋巴细胞与单核细胞/巨噬细胞）HIV 病毒载量。 将检查细胞亚区室随时间和区室之间的变化模式，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。 属于独特反应类别的患者 - 创造的不一致反应者（15% - 20% 的患者在治疗后病毒载量和 CD4 计数均增加，但似乎获得与预期病毒学反应的患者相同的临床益处），也将进行研究，以确定治疗反应的病理生理学参数，这些参数超出了吸收、依从性和表型/基因型耐药模式的范围，以解释 病毒学反应。 该项目将研究病毒向性的作用以及治疗方案在特定淋巴细胞亚型（包括终末分化细胞与分裂细胞）中的具体活性。 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，并将提供关于特定治疗方案如何影响总病毒载量的独特见解。 然而，该研究职业奖的首要目标是为 PI 提供与严密监督的临床研究期相重叠的教学培训，从而使 PI 成为一名可行的独立研究员。