Oral HIV Pathogenesis and Shedding

口腔艾滋病毒发病机制和脱落

• 批准号: 6947295
• 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 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6947295
• 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（由申请方提供）：HIV的主要感染部位是粘膜表面。虽然流行病学数据表明，口服艾滋病毒传播是罕见的，艾滋病毒可以在口腔分泌物中发现。在口咽部HIV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阳性男性，中位CD 4计数为356个细胞/穆尔; 52%接受了高效抗逆转录病毒治疗（HAART）。每周采集一次血液和口咽拭子，并评价HIV RNA。20名男性（29%）在舌和/或咽拭子中检出HIV RNA（vRNA）。广义估计方程显示血浆VL每增加1.0 log 10与咽部VL增加0.4 log 10相关（P<0.001），扁桃体切除术与咽部VL减少0.6 log 10相关（P = 0.007），但HAART与此无关（P > 0.1）。在20例vRNA+男性中，有5例（25%）从后口咽部分离出感染性HIV。培养阳性男性的中位粘膜VL为6.35 log 10拷贝/ml，而培养阴性男性为4.68 log 10拷贝/ml（P = 0.06）。未从唾液、口腔或唾液粘膜中分离出HIV。腭和舌扁桃体（n=6）活检显示vRNA和p24抗原淋巴滤泡内，尽管HAART抑制血浆VL。vRNA+细胞非常接近扁桃体囊并在扁桃体囊内迁移。HIV vRNA存在于扁桃体淋巴细胞、巨噬细胞（Mphi）和较少的树突状细胞（DC）中。粘多糖CCR 5+淋巴细胞是最普遍的vRNA+“dim”Mphi，通常携带> 10倍的vRNA和感染性病毒。口咽部病毒的检查显示出更大的env基因多样性比血液病毒同时检查，特别是从男性接受HAART（P = 0.004）。这些研究表明，origanyx是一个网站的艾滋病毒表达，其中Mphi似乎发挥了重要作用，在维持高滴度的复制艾滋病毒。先前存在的口腔病理和性行为，促进接触舌咽粘膜可能与病毒传播的风险增加。该提案致力于定义口咽部HIV感染的发病机制，并将重点确定支持HIV复制的解剖部位和细胞以及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，并描述了体外研究的组合，使用扁桃体外植体调查感染的动态，并在体内研究，以评估艾滋病毒的人口动态，可能是独特的口腔，包括耐药病毒的“区室化”。