IMMUNOTOXIN-DEPLETION OF HIV-1+T CELLS

描述：我们之前已经证明，一种免疫毒素(IT)指示 抗CD45RO，并含有化学去糖基化的蓖麻毒素A链 (DGA)，杀死潜伏和活跃的CD4阳性的人类T细胞 体外感染HIV-1。我们现在建议确定此IT是否可以 杀死HIV阳性个体中受感染的T细胞，特别是潜伏 来自接受HAART的个体的细胞。这项研究将涉及完善一个 高灵敏的极限稀释法测定血吸虫感染频率 具有复制能力的细胞。实验将包括体外实验和体内实验。 体内感染的CD4阳性T细胞感染耐药和 HIV-1的药物敏感分离株。如果IT有效地杀死了目标细胞， 然后我们将确定它对CD4阳性的免疫记忆有什么影响 和CD8阳性T细胞群。这将通过治疗来实现 HIV阴性和HIV阳性患者的外周血细胞 然后用有丝分裂原、超抗原和抗原激活细胞。 然后将使用流式细胞术来评估细胞内的细胞因子 幼稚和记忆性CD4阳性和CD8阳性T细胞亚群 确定T细胞内存是否受损。这是一项临床前研究 评估一种新的治疗方式。我们的结果应该决定 未来临床试验的可行性。

DESCRIPTION: We have previously demonstrated that an immunotoxin (IT) directed against CD45RO and containing chemically deglycosylated ricin toxin A chain (dgA), kills CD4-postive human T-cells which are both latently and actively infected with HIV-1 in vitro. We now propose to determine whether this IT can kill infected T-cells from HIV positive individuals and, in particular, latent cells from individuals receiving HAART. This study will involve perfecting a highly sensitive limiting dilution assay to determine the frequency of replication-competent cells. Experiments will include both in vitro- and in vivo-infected CD4-postive T cells infected with drug-resistant and drug-sensitive isolates of HIV-1. If the IT effectively kills the target cells, we will then determine what effect it has on immune memory in the CD4-positive and CD8-positive T-cell populations. This will be accomplished by treating peripheral blood cells from both HIV-negative and HIV-positive individuals with the IT and then activating cells with mitogens, superantigens, and antigens. Flow cytometry will then be used to evaluate intracellular cytokines in sub-populations of naive and memory CD4-postive and CD8-postive T-cells to determine whether T cell memory is compromised. This is a preclinical study to evaluate a new therapeutic modality. Our results should determine the feasibility of a future clinical trial.