Phenotypic analysis of latently infected CD4+ T cells.

产品说明：人们对利用小分子重新激活潜伏感染细胞等策略根除HIV-1的可能性持乐观态度。最近的一项研究表明，这种方法实际上是可能的;单剂量的伏立诺他分子显示出在体内诱导CD 4 + T细胞中的HIV表达。虽然这种策略最终可能是有效的，但重新激活每一个潜伏感染的CD 4 + T细胞可能非常具有挑战性。 补充策略可以是使用与毒素缀合的抗体物理地消除潜伏感染的细胞。这可能类似于在开始化疗之前手术“减积”肿瘤。然而，靶向潜伏感染的CD 4 + T细胞一直是一项具有挑战性的任务，因为这些细胞的表型在很大程度上仍然未知。在这项新的提议中，我们计划使用针对200多种不同T细胞标志物的抗体，以确定潜伏感染的CD 4 + T细胞的表型。在最近的一项研究中，这种方法已成功地用于确定活化T细胞的表型。这将类似于筛选活化潜伏感染的CD 4 + T细胞的小分子，但在这种情况下，将使用单克隆抗体（Mab）而不是化合物的文库，并且读出将是Mab与感染的静息CD 4 + T细胞或最近再活化的潜伏感染的CD 4 + T细胞的结合，而不是与CD 4 + T细胞结合。 重新激活自己。我们计划通过对接受抑制性HAART方案的HIV-1感染患者的静息CD 4 + T细胞进行分选来验证我们的结果。我们将专门对筛选中识别的标记进行分类，以确定这些细胞中是否存在HIV-1 DNA和潜伏复制能力病毒的过度表达。 这项工作将对HIV-1根除策略产生重大影响，因为它可能导致潜在感染细胞的清除。

DESCRIPTION: There has been a lot of optimism about potentially eradicating HIV-1 using strategies such as reactivation of latently infected cells by small molecules. A recent study has suggested that this approach may in fact be possible; a single dose of the molecule vorinostat was shown to induce HIV expression in CD4+ T cells in vivo. While this strategy may end up being effective, it may be very challenging to reactivate every last latently infected CD4+ T cell. A complementary strategy may be to physically eliminate latently infected cells using antibodies conjugated to toxins. This may be analogous to surgically "debulking" a tumor prior to initiating chemotherapy. However, targeting latently infected CD4+ T cells has been a challenging task because the phenotype of these cells remains largely unknown. In this new proposal, we plan to use antibodies to more than 200 different T cell markers in order to determine the phenotype of latently infected CD4+ T cells. Such an approach has been successfully used to determine the phenotype of activated T cells in a recent study. This will be analogous to screening for small molecules that activate latently infected CD4+ T cells, but in this case, a library of monoclonal antibodies (Mabs) rather than compounds will be used and the readout will be binding of Mabs to infected resting CD4+ T cells or recently reactivated latently infected CD4+ T cells rather than reactivation itself. We plan to validate our results by sorting resting CD4+ T cells from HIV-1 infected patients on suppressive HAART regimens. We specifically will sort for markers that are identified in our screen to determine whether there is an overrepresentation of HIV-1 DNA and latent replication-competent virus in these cells. This work will have major implications for strategies for HIV-1 eradication since it will potentially lead to the clearance of latently infectd cells.