HIV-1-Specific Chimeric Antigen Receptor T Cells Fail To Recognize and Eliminate the Follicular Dendritic Cell HIV Reservoir In Vitro.
HIV-1-Specific Chimeric Antigen Receptor T Cells Fail To Recognize and Eliminate the Follicular Dendritic Cell HIV Reservoir In Vitro.
复制标题
HIV-1 特异性嵌合抗原受体 T 细胞无法在体外识别并消除滤泡树突状细胞 HIV 储库。
DOI：
10.1128/jvi.00190-20
复制
发表时间：
2020
影响因子：
5.4
通讯作者：
Burton,GregoryF
中科院分区：
文献类型：
--
作者：
Ollerton,MatthewT;Berger,EdwardA;Connick,Elizabeth;Burton,GregoryF
治愈艾滋病毒感染的主要障碍是在抗逆转录病毒治疗期间持续存在具有复制能力的病毒库。HIV特异性嵌合抗原受体(CAR)T细胞已被开发用于靶向潜伏感染的CD4+T细胞，这些细胞自发地或在故意潜伏逆转后表达病毒。HIV特异性CAR-T细胞是否能够识别和清除HIV结合免疫复合体(IC)的滤泡树突状细胞(FDC)储存库尚不清楚。我们使用人外周血单个核细胞(PBMC)和一个能够表达CD4(结构域1和2)和甘露糖结合凝集素(MBL)的碳水化合物识别结构域(CD4-MBL CAR)的CAR-T细胞来创建HIV特异性T细胞。用羧基荧光素琥珀酰亚胺酯释放法检测CAR-T细胞的细胞毒作用，用流式细胞仪检测干扰素-γ的产生和CD107a膜积聚来评价CAR-T细胞的活化。CD4-MBL CAR-T细胞对表达Env的细胞系和HIV感染的CD4+T细胞表现出较强的裂解和功能性反应，但对携带HIV-IC的FDC无效。在无细胞实验中，CD4-MBL CAR-T细胞对无细胞HIV或浓缩、固定化的HIV-IC无反应。封闭细胞间黏附分子-1(ICAM-1)抑制了CD4-MBL CAR-T细胞对表达Env的细胞系和HIV感染的CD4+T细胞的杀伤反应，提示黏附分子等因素是稳定CAR-Env相互作用所必需的，从而引发细胞毒反应。因此，CD4-MBL CAR-T细胞不能消除与FDC相关的HIV储存库，必须采取替代策略来根除这个储存库。治疗HIV感染的主要重点是消除潜伏感染的CD4+T细胞。很少有研究涉及存在于滤泡树突状细胞(FDCs)上的独特的感染性艾滋病毒储存库，它在抗逆转录病毒治疗期间持续存活，并可能在抗逆转录病毒治疗停止后导致病毒反弹。我们评估了一种新型的HIV特异性嵌合抗原受体(CAR)T细胞在体外靶向HIV感染的CD4+T细胞和FDC储存库的有效性。虽然CAR-T细胞清除了表达HIV的CD4+T细胞，但它们对与HIV结合的FDC没有反应或消除。这些发现揭示了CAR-T细胞疗法根除艾滋病毒的根本局限性。
The major obstacle to a cure for HIV infection is the persistence of replication-competent viral reservoirs during antiretroviral therapy. HIV-specific chimeric antigen receptor (CAR) T cells have been developed to target latently infected CD4+T cells that express virus either spontaneously or after intentional latency reversal. Whether HIV-specific CAR-T cells can recognize and eliminate the follicular dendritic cell (FDC) reservoir of HIV-bound immune complexes (ICs) is unknown. We created HIV-specific CAR-T cells using human peripheral blood mononuclear cells (PBMCs) and a CAR construct that enables the expression of CD4 (domains 1 and 2) and the carbohydrate recognition domain of mannose binding lectin (MBL) to target native HIV Env (CD4-MBL CAR). We assessed CAR-T cell cytotoxicity using a carboxyfluorescein succinimidyl ester (CFSE) release assay and evaluated CAR-T cell activation through interferon gamma (IFN-γ) production and CD107a membrane accumulation by flow cytometry. CD4-MBL CAR-T cells displayed potent lytic and functional responses to Env-expressing cell lines and HIV-infected CD4+T cells but were ineffective at targeting FDC bearing HIV-ICs. CD4-MBL CAR-T cells were unresponsive to cell-free HIV or concentrated, immobilized HIV-ICs in cell-free experiments. Blocking intercellular adhesion molecule-1 (ICAM-1) inhibited the cytolytic response of CD4-MBL CAR-T cells to Env-expressing cell lines and HIV-infected CD4+T cells, suggesting that factors such as adhesion molecules are necessary for the stabilization of the CAR-Env interaction to elicit a cytotoxic response. Thus, CD4-MBL CAR-T cells are unable to eliminate the FDC-associated HIV reservoir, and alternative strategies to eradicate this reservoir must be sought.IMPORTANCEEfforts to cure HIV infection have focused primarily on the elimination of latently infected CD4+T cells. Few studies have addressed the unique reservoir of infectious HIV that exists on follicular dendritic cells (FDCs), persistsin vivoduring antiretroviral therapy, and likely contributes to viral rebound upon cessation of antiretroviral therapy. We assessed the efficacy of a novel HIV-specific chimeric antigen receptor (CAR) T cell to target both HIV-infected CD4+T cells and the FDC reservoirin vitro. Although CAR-T cells eliminated CD4+T cells that express HIV, they did not respond to or eliminate FDC bound to HIV. These findings reveal a fundamental limitation to CAR-T cell therapy to eradicate HIV.