EVALUATION OF RANTES ANALOGUES FOR PROTECTION FROM SHIV

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 迫切需要有效的艾滋病毒预防战略，但最近关键疫苗和“杀微生物剂”临床试验的失败突显了在相关动物模型中验证新方法的必要性。然而，我们现在知道CCR5是病毒融合的主要受体，RANTES和RANTES类似物可以结合CCR5，使其无法与病毒结合。因此，我们一直在开发RANTES类似物作为杀微生物剂的候选者。然而，Achour等人最近的一份报告。提示原代T细胞内存在CCR5池，如果受体在阻断后迅速重新表达，这种方法可能会使这种方法变得无用。通过使用一系列互补的技术来检测CCR5的表达(抗体标记、Western印迹、qRT-PCR)，我们证实了CCR5的胞内池不存在，并证实了Achour等人的结果。假阳性是由于在细胞固定和通透性过程中产生了与抗CCR5抗体无关的结合部位而产生的。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Effective strategies for HIV prevention are urgently needed, but recent failures in key vaccine and 'microbicide' clinical trials highlight the need for new approaches validated in relevant animal models. However we now know that CCR5 is the major receptor for viral fusion and that RANTES and RANTES analogs can bind CCR5, making it inaccessible for viruses. Thus we have been developing RANTES analogs as microbicide candidates. However a recent report by Achour et al. suggested that primary T cells harbor pools of intracellular CCR5 which could render such an approach useless if receptors were rapidly re-expressed after blockage. By using a series of complementary techniques to measure CCR5 expression (antibody labeling, Western blot, qRT-PCR), we established that intracellular pools of CCR5 do not exist and that the results obtained by Achour et al. were false-positives that arose due to the generation of irrelevant binding sites for anti-CCR5 antibodies during fixation and permeabilization of cells.