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一次性治疗持久缓解的方法 在没有抗逆转录病毒药物的情况下，人类免疫缺陷病毒(艾滋病毒)的感染。这种疗法是一种 靶向HIV特异性嵌合抗原受体(CAR，特别是CD4-MBL-CAR)T细胞治疗 B细胞滤泡1、2.B细胞滤泡是一种免疫保护部位，由于低水平的 病毒特异性CD8 T细胞3-6。我们的临床前CAR T细胞在猴免疫缺陷病毒(SIV)中的初步研究- 恒河猴HIV感染模型表明，该免疫疗法是安全有效的。在这里，我们 建议开发和评估一种类似的人类T细胞免疫疗法。病毒特异性CD8 T细胞发挥作用 在体外和体内对HIV-1和SIV具有很强的抗病毒活性。然而，尽管CD8 T细胞数量丰富 HIV-1感染者和SIV感染猕猴的细胞反应，这些细胞无法完全抑制 病毒复制。这种不充分的抑制很可能是因为大多数HIV-1和SIV 复制发生在次级淋巴组织中集中在B细胞滤泡内的CD4T细胞7-11，其中 相对较少的病毒特异性CD8T细胞残留3-6。事实上，我们在体内证明了效靶比 效应病毒特异性CD8T细胞对SIV RNA细胞的靶向比B细胞外低40倍 SIV感染猕猴期间淋巴组织中的滤泡5。此外，我们还透露，大多数人 的病毒特异性CD8 T细胞不表达毛囊归巢分子CXCR55，这可能解释了低水平的 病毒特异性CD8 T细胞定位于B细胞滤泡并对其进行监测。这些数据表明， HIV和SIV特异性CD8T细胞完全抑制病毒复制可能是由于缺乏病毒特异性 B细胞滤泡中的CD8T细胞。基于这些发现和我们临床前灵长类研究的成功，我们 建议开发一种HIV特异性的CD4-MBL-CAR T细胞疗法，该疗法采用卵泡归巢 分子CXCR5引导HIV特异性T细胞向B细胞滤泡迁移。我们特别建议 目的：1)培养人CD4-MBL-CAR/CXCR5 T细胞；2)确定人T细胞的表型 CD4-MBL-CAR/CXCR5 T细胞，3)决定人CD4-MBL-CAR/CXCR5 T细胞的功能。我们的 开发定位于淋巴组织并在淋巴组织中发挥功能的人CD4-MBL-CAR/CXCR5 T细胞的拟议研究 在没有抗逆转录病毒药物的情况下，B细胞滤泡可能会导致艾滋病毒感染的持久缓解。

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.