Development of a treatment for the durable remission of HIV using autologous human CAR T cells that target B cell follicles

摘要 在我们成功的灵长类动物模型研究的基础上，我们的目标是开发一种持久缓解的一次性治疗方法 在没有抗逆转录病毒药物的情况下，艾滋病毒的感染。这种治疗是一种 自体HIV特异性嵌合抗原受体（CAR，特别是CD 4-MBL-CAR）T细胞疗法，其靶向 B细胞滤泡1，2. B细胞滤泡是一个免疫保护位点，由于低水平的免疫抑制， 病毒特异性CD 8 T细胞3 -6。我们在猴免疫缺陷病毒（SIV）中的临床前CAR T细胞试验研究- 恒河猴HIV感染模型的研究表明，该免疫疗法是安全有效的。这里我们 建议开发和评估一种可比较的人类T细胞免疫疗法。病毒特异性CD 8 T细胞发挥 在体外和体内对HIV-1和SIV都具有有效的抗病毒活性。尽管有大量的CD 8 T细胞， 在HIV-1感染的人类和SIV感染的猕猴中，这些细胞无法完全抑制 病毒复制这种不充分的抑制可能是由于大多数HIV-1和SIV 复制发生在次级淋巴组织的B细胞滤泡内集中的CD 4 + T细胞7 -11中， 相对较少的病毒特异性CD 8 T细胞残基3 -6。事实上，我们发现，在体内， 与B细胞外部相比，靶向SIV RNA+细胞的效应病毒特异性CD 8 T细胞内部低>40倍 恒河猴SIV感染过程中淋巴组织中的滤泡5.此外，我们发现，大多数 的病毒特异性CD 8 T细胞不能表达滤泡归巢分子CXCR 55，这可能解释了低水平的 病毒特异性CD 8 T细胞定位于并监视B细胞滤泡。这些数据表明， HIV和SIV特异性CD 8 T细胞完全抑制病毒复制可能是由于缺乏病毒特异性T细胞。 B细胞滤泡中的CD 8 T细胞。基于这些发现和我们的临床前灵长类动物研究的成功，我们 我建议开发一种HIV特异性CD 4-MBL-CAR T细胞疗法， 分子CXCR 5来指导HIV特异性T细胞向B细胞滤泡的迁移。我们特别建议 本研究的目的是：1）开发人CD 4-MBL-CAR/CXCR 5 T细胞，2）确定人CD 4-MBL-CAR/CXCR 5 T细胞的表型， ⑶ 4-MBL-CAR/CXCR 5 T细胞，3）测定人⑶ 4-MBL-CAR/CXCR 5 T细胞的功能。我们 提出的研究开发人CD 4-MBL-CAR/CXCR 5 T细胞，其定位于淋巴样细胞并在淋巴样细胞中起作用。 B细胞滤泡可能导致在没有抗逆转录病毒药物的情况下HIV感染的持久缓解。

Abstract Building on our successful primate model studies, we aim to develop a one-time treatment for durable remission of human immunodeficiency virus (HIV) in the absence of antiretroviral medications. This treatment is an autologous HIV-specific chimeric antigen receptor (CAR, specifically CD4-MBL-CAR) T cell therapy that targets B cell follicles1,2. B cell follicles are an immune protected site that permit viral replication due to low levels of virus-specific CD8 T cells3–6. Our preclinical CAR T cell pilot studies in a simian immunodeficiency virus (SIV)- infected rhesus macaque model of HIV indicated that this immunotherapy is safe and effective. Here, we propose to develop and evaluate a comparable human T cell immunotherapy. Virus-specific CD8 T cells exert potent antiviral activity against HIV-1 and SIV both in vitro and in vivo. Nevertheless, despite abundant CD8 T cell responses in HIV-1-infected humans and SIV-infected macaques, these cells are unable to fully suppress virus replication. This inadequate suppression is likely due to the fact that the majority of HIV-1 and SIV replication occurs in CD4+ T cells7–11 concentrated within B cell follicles in secondary lymphoid tissues, where relatively few virus-specific CD8 T cells reside3–6. In fact, we showed that the effector to target ratio of in vivo effector virus-specific CD8 T cell to target SIV RNA+ cells is >40-fold lower inside compared to outside of B cell follicles in lymphoid tissues during SIV infection in rhesus macaque5. Furthermore, we revealed that the majority of virus-specific CD8 T cells fail to express the follicular homing molecule CXCR55, likely explaining low levels of virus-specific CD8 T cells localizing to and surveilling B cell follicles. These data suggest that the inability of HIV- and SIV-specific CD8 T cells to fully suppress virus replication may be due to a deficiency of virus-specific CD8 T cells in B cell follicles. Based on these findings and the success of our preclinical primate studies, we propose to develop an HIV-specific CD4-MBL-CAR T cell therapy that employs the follicular homing molecule CXCR5 to direct the migration of HIV-specific T cells to B cell follicles. We specifically propose the following aims, 1) Develop human CD4-MBL-CAR/CXCR5 T cells, 2) Determine the phenotype of human CD4-MBL-CAR/CXCR5 T cells, 3) Determine the function of human CD4-MBL-CAR/CXCR5 T cells. Our proposed studies to develop human CD4-MBL-CAR/CXCR5 T cells that localize to and function in lymphoid B cell follicles may lead to durable remission of HIV infection in the absence of antiretroviral drugs.