Oral HIV Pathogenesis and Shedding

口腔艾滋病毒发病机制和脱落

• 批准号: 6659013
• 负责人: Robert W. Coombs
• 金额: $ 33.31万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6659013
• 关键词: AIDS education /prevention; HIV infections; clinical research; communicable disease transmission; counseling; drug resistance; helper T lymphocyte; human subject; immunocytochemistry; immunopathology; in situ hybridization; interview; macrophage; mucosa; oral health; oral pharyngeal; pathologic process; polymerase chain reaction; tissue /cell culture; virus RNA; virus load; virus replication

DESCRIPTION (provided by applicant): The primary sites of acquisition of HIV are at mucosal surfaces. Although epidemiological data suggests oral HIV transmission is infrequent, HIV can be found in oral secretions. In a preliminary study of oropharyngeal HIV shedding, we examined 70 HIV+ men from Seattle and Peru, with a median CD4 count of 356 cells/mul; 52% were receiving highly active anti-retroviral therapy (HAART). Blood and oropharyngeal swabs were taken at weekly intervals and evaluated for HIV RNA. 20 men (29%) had detectable HIV RNA (vRNA) in lingual and/or pharyngeal swabs. Generalized estimating equations were used to show that each 1.0 log10 increase in plasma VL was associated with a 0.4 log10 increase in pharyngeal VL (P<0.001), and tonsillectomy was associated with a 0.6 log10 reduction in pharyngeal VL (P = 0.007), but HAART was not (P > 0.1). Infectious HIV was isolated from the posterior oropharynx in 5 (25%) of 20 vRNA+ men. Median mucosal VL was 6.35 log10 copies/ml in culture-positive men vs. 4.68 log10 copies/ml in culture-negative men (P = 0.06). HIV was not isolated from saliva, buccal or salivary mucosa. Biopsies of palatine and lingual tonsil (n=6) revealed vRNA and p24 antigen within lymphoid follicles, despite HAART-suppressed plasma VL. vRNA+ cells were in close proximity to and migrating within the tonsil capsule. HIV vRNA was present within tonsilar lymphocytes, macrophages (Mphi) and fewer dendritic cells (DC). Mucosal CCR5+ lymphocytes were the most prevalent vRNA+ 'dim' Mphi often harbored > 10-fold more vRNA and infectious virus. An examination of oropharyngeal viruses revealed greater env gene diversity than blood viruses examined simultaneously, particularly from men receiving HAART (P = 0.004). These studies demonstrate that the oropharyx is a site of HIV expression, where Mphi appear to play an important role in maintaining high titers of replicative HIV. Pre-existing oral pathology and sexual acts that facilitate contact with the lingual-pharyngeal mucosa may be associated with increased risk of viral transmission. This proposal is devoted to defining the pathogenesis of HIV infection in the oropharynx and will focus on determining anatomic site(s) and cells that support HIV replication and factors that influence the frequency and titer of virus at mucosal surfaces. It employs state-of-the-art methodologies and describes a combination of in vitro studies, using tonsil explants to investigate the dynamics of infection, and in vivo studies to assess HIV population dynamics that may be unique to the oral cavity, including 'compartmentalization' of drug-resistant virus.

描述（由申请人提供）：艾滋病毒收购的主要地点位于粘膜表面。尽管流行病学数据表明口服HIV传播很少，但在口腔分泌中可以发现HIV。在对口咽艾滋病毒脱落的初步研究中，我们检查了来自西雅图和秘鲁的70名HIV+男性，中位CD4计数为356个细胞/mul； 52％的人接受了高度活跃的抗逆转录病毒疗法（HAART）。每周抽取血液和口咽拭子，并评估HIV RNA。有20名男性（29％）在脑脊髓和/或咽拭子中具有可检测的HIV RNA（VRNA）。使用广义估计方程来表明，等离子体VL中的每1.0 Log10增加与咽VL的0.4 log10相关（p <0.001），扁桃体切除术与咽VL的0.6 log10降低有关（p = 0.007），但是哈特不是（p> 0.1）。在20个VRNA+男性中有5个（25％）中，从后口腔中分离出感染性HIV。在培养阳性男性中，中位粘膜VL为6.35 log10副本/ml，培养文化男性中的4.68 log10副本/ml（p = 0.06）。 HIV不是从唾液，颊或唾液粘膜中分离出来的。尽管Haart抑制了血浆VL，但palatine和舌扁桃体（n = 6）的活检显示淋巴卵泡中的VRNA和P24抗原。 VRNA+细胞在扁桃体胶囊内靠近和迁移。 HIV VRNA存在于Tonsilar淋巴细胞，巨噬细胞（MPHI）和较少的树突状细胞（DC）中。粘膜CCR5+淋巴细胞是最普遍的VRNA+'DIM'MPHI，通常具有> 10倍的VRNA和感染性病毒。对口咽病毒的检查发现，与同时研究的血液病毒相比，ENV基因的多样性更大，特别是从接受HAART的男性（P = 0.004）。这些研究表明，口咽是HIV表达的部位，其中MPHI似乎在维持较高的复制HIV滴度中起着重要作用。促进与舌咽粘膜接触的先前的口腔病理和性行为可能与病毒传播的风险增加有关。该建议致力于定义口咽中HIV感染的发病机理，并将着重于确定支持HIV复制的解剖部位和细胞，这些细胞支持HIV复制和影响粘膜表面病毒频率和滴度的因素。它采用了最先进的方法论，并描述了体外研究的组合，使用扁桃体外植体研究感染的动力学以及体内研究，以评估可能是口腔腔所特有的HIV种群动态，包括'carptertallization carpmentalization '耐药病毒。