Defining HIV and SIV Reservoirs in Lymphoid Tissues.
Defining HIV and SIV Reservoirs in Lymphoid Tissues.
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DOI：
10.20411/pai.v1i1.100
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发表时间：
2016
影响因子：
--
通讯作者：
Estes JD
中科院分区：
文献类型：
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作者：
Deleage C;Wietgrefe SW;Del Prete G;Morcock DR;Hao XP;Piatak M Jr;Bess J;Anderson JL;Perkey KE;Reilly C;McCune JM;Haase AT;Lifson JD;Schacker TW;Estes JD
治愈HIV-1的主要障碍是长寿的病毒储库，必须消除或大大减少。治疗策略主要集中在监测外周血（PB）中T细胞储库的变化，尽管淋巴组织（LT）是病毒持续存在的主要部位。为了追踪和区分组织隔室内的病毒储库，我们开发了特异性和灵敏性的下一代原位杂交方法来检测vRNA，包括vRNA+细胞和病毒颗粒（“RNAscope”）、vDNA+细胞（“DNAscope”）以及组合的vRNA和vDNA与免疫组织化学，以检测和表型相同组织切片中的活性和潜伏感染的细胞。与传统的原位杂交相比，RNAscope具有高灵敏度和更快的分析速度。高灵敏度、高特异性的DNAscope可检测SIV/HIV vDNA+细胞，包括vDNA和vRNA或免疫表型标记在同一切片上的重叠检测。在联合抗逆转录病毒治疗之前和期间对来自猕猴的LT样品的分析表明，B细胞滤泡是治疗期间潜伏和活性病毒持续存在的重要解剖隔室。这些新工具应该允许对病毒储库生物学和治疗策略的评估有新的见解。
A primary obstacle to an HIV-1 cure is long-lived viral reservoirs, which must be eliminated or greatly reduced. Cure strategies have largely focused on monitoring changes in T cell reservoirs in peripheral blood (PB), even though the lymphoid tissues (LT) are primary sites for viral persistence. To track and discriminate viral reservoirs within tissue compartments we developed a specific and sensitive next-generation in situ hybridization approach to detect vRNA, including vRNA+ cells and viral particles (“RNAscope”), vDNA+ cells (“DNAscope”) and combined vRNA and vDNA with immunohistochemistry to detect and phenotype active and latently infected cells in the same tissue section. RNAscope is highly sensitive with greater speed of analysis compared to traditional in situ hybridization. The highly sensitive and specific DNAscope detected SIV/HIV vDNA+ cells, including duplexed detection of vDNA and vRNA or immunophenotypic markers in the same section. Analysis of LT samples from macaques prior to and during combination antiretroviral therapy demonstrated that B cell follicles are an important anatomical compartment for both latent and active viral persistence during treatment. These new tools should allow new insights into viral reservoir biology and evaluation of cure strategies.