Bioengineering T cells to target carbohydrate to treat opportunistic fungal infection
Bioengineering T cells to target carbohydrate to treat opportunistic fungal infection
复制标题
DOI：
10.1073/pnas.1312789111
复制
发表时间：
2014-07-22
影响因子：
11.1
通讯作者：
Cooper, Laurence J. N.
中科院分区：
文献类型：
--
作者：
Kumaresan, Pappanaicken R.;Manuri, Pallavi R.;Cooper, Laurence J. N.
临床级T细胞经离体遗传修饰以表达嵌合抗原受体（汽车），从而将其特异性重定向至体内靶向肿瘤相关抗原。我们现在已经开发出这种分子策略，使细胞毒性T细胞特异性针对真菌。我们使模式识别受体Dectin-1适应于在与曲霉属胚细胞的细胞壁中的碳水化合物结合后通过嵌合的CD 28和CD 3-zeta（命名为“D-CAR”）激活T细胞。使用类似于食品和药物管理局批准的用于制造用于临床试验的CD 19特异性CAR（+）T细胞的方法，在人工活化和繁殖细胞上选择性地繁殖用睡美人系统遗传修饰以稳定表达D-CAR的T细胞。D-CAR（+）T细胞表现出对β-葡聚糖的特异性，这导致体外和体内曲霉菌菌丝生长的损伤和抑制。用类固醇处理D-CAR（+）T细胞并没有显著损害抗真菌活性。这些数据支持CAR（+）T细胞靶向碳水化合物抗原，并提供了一种临床上有吸引力的策略，以使用T细胞基因疗法增强对机会性真菌感染的免疫力。
Clinical-grade T cells are genetically modified ex vivo to express chimeric antigen receptors (CARs) to redirect their specificity to target tumor-associated antigens in vivo. We now have developed this molecular strategy to render cytotoxic T cells specific for fungi. We adapted the pattern-recognition receptor Dectin-1 to activate T cells via chimeric CD28 and CD3-zeta (designated "D-CAR") upon binding with carbohydrate in the cell wall of Aspergillus germlings. T cells genetically modified with the Sleeping Beauty system to express D-CAR stably were propagated selectively on artificial activating and propagating cells using an approach similar to that approved by the Food and Drug Administration for manufacturing CD19-specific CAR(+) T cells for clinical trials. The D-CAR(+) T cells exhibited specificity for beta-glucan which led to damage and inhibition of hyphal growth of Aspergillus in vitro and in vivo. Treatment of D-CAR(+) T cells with steroids did not compromise antifungal activity significantly. These data support the targeting of carbohydrate antigens by CAR(+) T cells and provide a clinically appealing strategy to enhance immunity for opportunistic fungal infections using T-cell gene therapy.