In vitro immune modulation by antibodies coupled to tumour cells
In vitro immune modulation by antibodies coupled to tumour cells
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DOI：
10.1038/sj.gt.3300534
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发表时间：
1997-12-01
期刊：
影响因子：
5.1
通讯作者：
Farzaneh, F
中科院分区：
文献类型：
--
作者：
Darling, D;GaleaLauri, J;Farzaneh, F
修饰自体肿瘤细胞以表达免疫共刺激剂 B7.1 是癌症免疫治疗的潜在策略。此前，这涉及将遗传物质引入细胞、体外培养以及细胞表面蛋白质产物的确认。只有在短时间内获得足够的肿瘤并进行有效修饰，这才有可能。虽然离体肿瘤细胞培养和转染/感染程序已取得进展，但目前的基因转移手段对于某些肿瘤类型仍然不够有效。我们描述了一种高效的体外程序，可以修饰群体中 99% 以上的细胞，从而表达具有潜在免疫调节活性的细胞表面蛋白。该过程可在短短 24 小时内完成，对细胞数量没有上限，利用琥珀酰亚胺酯用生物素共价标记细胞表面蛋白。然后生物素化的细胞膜蛋白锚定并形成亲和素桥以固定蛋白 G-生物素。这可以通过 Fc 区结合免疫球蛋白 (Ig) 分子，从而使抗体的可变区自由且功能可用。在本研究中，刺激性小鼠抗人 CD28 单克隆抗体与肿瘤细胞表面的结合用于表明修饰的细胞能够在体外共刺激 T 细胞。该方法的简单性以及常用试剂的使用，代表着向现实的、真正的“现成的”非基因免疫治疗方案又迈出了一步。
Modification of autologous tumour cells to express the immune costimulator B7.1 is a potential strategy for immunotherapy of cancer. Previously, this has involved introduction of genetic material into cells, in vitro culture, and confirmation of the protein product on the cell surface. This is possible only if sufficient tumour is obtainable and efficiently modified in a short time. Whilst progress has been made on ex vivo tumour cell culture and transfection/infection procedures there are still tumour types for which the present means of gene transfer are not efficient enough. We describe a highly efficient in vitro procedure for the modification of over 99% of the cells in a population, allowing the expression of cell surface proteins with potential immune modulatory activities. This procedure, which can be completed in as little as 24 h with no upper limit on cell number, utilizes succinimide ester to label cell surface proteins with biotin covalently. Biotinylated cell membrane proteins then anchor and avidin bridge for immobilizing protein G-biotin. This can serve to bind immunoglobulin (Ig) molecules via their Fc region such that the variable region of the antibody is freely and functionally available. In the present study th binding of a stimulatory mouse anti-human CD28 monoclonal antibody to the surface of tumour cells is used to show that the modified cells are capable of co-stimulating T cells in vitro. The simplicity of the method, and the use of common reagents, represents a further step towards a realistic, truly 'off-the-shelf', nongene immunotherapy protocol.