INSERM U1040, Montpellier, France.
Haematologica. 2011 Mar;96(3):400-7. doi: 10.3324/haematol.2010.029520. Epub 2010 Nov 25.
Anti-CD20 monoclonal antibodies are major therapeutic agents for patients with follicular lymphoma and work through complement-mediated cytotoxicity and antibody-dependent cellular cytotoxicity. Optimization of antibody-dependent cellular cytotoxicity, in particular by amplifying its effectors, could further increase the efficacy of anti-CD20 monoclonal antibodies.
We investigated the cytotoxic activity of Vγ9Vδ2 T cells against follicular lymphoma cells and whether this killing could be increased by promoting antibody-dependent cellular cytotoxicity with anti-CD20 monoclonal antibodies, in particular a type-II glycoengineered anti-CD20. Vγ9Vδ2 T cells were expanded in vitro in the presence of bromohydrin pyrophosphate (Phosphostim) and interleukin-2 and their ability to kill follicular lymphoma primary cells or cell lines was evaluated by flow cytometry cytotoxic T-lymphocyte assays in the presence or absence of three anti-CD20 monoclonal antibodies: the afucosylated GA101, the chimeric rituximab or the humanized ofatumumab. The ability of these cells to release perforin/granzyme and secrete interferon-γ when co-cultured with follicular lymphoma primary cells or cell lines in the presence or not of the three anti-CD20 monoclonal antibodies was also evaluated by CD107a staining and Elispot assays.
Phosphostim and interleukin-2 expanded Vγ9Vδ2 T cells were cytotoxic to primary follicular lymphoma cells and their cytotoxic potential was dramatically increased by GA101, a type II glycoengineered anti-CD20 monoclonal antibody, and to a lesser extent, by rituximab and ofatumumab. The increased cytotoxicity was associated with increased secretion of perforin/granzyme and interferon-γ.
In-vitro expanded Vγ9Vδ2 T cells efficiently kill primary follicular lymphoma cells and express CD16; anti-CD20 monoclonal antibodies, in particular GA101, dramatically increase the cytotoxic activity of expanded Vγ9Vδ2 T cells. These preclinical results prompt the development of clinical trials using this antibody dependent cellular cytotoxicity property of Vγ9Vδ2 T cells and anti-CD20 monoclonal antibodies.
抗 CD20 单克隆抗体是滤泡性淋巴瘤患者的主要治疗药物，通过补体介导的细胞毒性和抗体依赖性细胞毒性发挥作用。通过扩增抗体依赖性细胞毒性的效应物，优化抗体依赖性细胞毒性，可以进一步提高抗 CD20 单克隆抗体的疗效。
我们研究了 Vγ9Vδ2 T 细胞对滤泡性淋巴瘤细胞的细胞毒性作用，以及通过用抗 CD20 单克隆抗体，特别是 II 型糖基化工程抗 CD20 增强抗体依赖性细胞毒性是否可以增加这种杀伤作用。Vγ9Vδ2 T 细胞在溴代二磷酸氢盐（Phosphostim）和白细胞介素 2 的存在下体外扩增，并通过流式细胞术细胞毒性 T 淋巴细胞试验评估其杀伤滤泡性淋巴瘤原代细胞或细胞系的能力，该试验在存在或不存在三种抗 CD20 单克隆抗体的情况下进行：去岩藻糖基化的 GA101、嵌合型利妥昔单抗或人源化奥法妥珠单抗。当与滤泡性淋巴瘤原代细胞或细胞系共培养时，还通过 CD107a 染色和 Elispot 试验评估这些细胞在存在或不存在三种抗 CD20 单克隆抗体的情况下释放穿孔素/颗粒酶和分泌干扰素-γ的能力。
Phosphostim 和白细胞介素 2 扩增的 Vγ9Vδ2 T 细胞对原代滤泡性淋巴瘤细胞具有细胞毒性，GA101（一种 II 型糖基化工程抗 CD20 单克隆抗体）显著增强了其细胞毒性潜能，而利妥昔单抗和奥法妥珠单抗的增强作用则较小。增加的细胞毒性与穿孔素/颗粒酶和干扰素-γ的分泌增加有关。
体外扩增的 Vγ9Vδ2 T 细胞有效地杀伤原代滤泡性淋巴瘤细胞，并表达 CD16；抗 CD20 单克隆抗体，特别是 GA101，显著增强了扩增的 Vγ9Vδ2 T 细胞的细胞毒性活性。这些临床前结果促使开发利用 Vγ9Vδ2 T 细胞的抗体依赖性细胞毒性和抗 CD20 单克隆抗体的临床试验。
