McHugh R S, Nagarajan S, Wang Y C, Sell K W, Selvaraj P
Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Cancer Res. 1999 May 15;59(10):2433-7.
Modification of tumor cells with one or more costimulatory adhesion molecules has been proposed as a means to develop therapeutic cancer vaccines for use in human immunotherapy. Expression of B7-1 (CD80) in tumors by gene transfer creates an immunogenic tumor cell that induces antitumor immunity and protects mice from further challenge with wild-type tumor cells. In this report, we demonstrate that protein transfer of glycosyl-phosphatidylinositol (GPI)-anchored costimulatory molecules into tumor cell membranes could be used as an alternative to gene transfer for tumor immunotherapy. Incubation of isolated tumor membranes with purified GPI-anchored B7-1 results in stable incorporation of B7-1 on tumor cell membranes within a few hours. Immunization of C57BL/6 mice with EG7 tumor membranes modified to express GPI-B7-1 by protein transfer induces tumor-specific T-cell proliferation and CTLs. In addition, immunization with these EG7 membranes protects mice from parental tumor challenge. The protein transfer approach used here does not require foreign vectors or live tumor cells and is completed within a matter of hours. Irradiated cells or membrane preparations from fresh or frozen tumor tissue can be used. Therefore, protein transfer of glycolipid-anchored molecules provides an efficient and novel approach to modify tumor membranes for human immunotherapy. This approach is not limited to costimulatory molecules because any cell surface protein can be converted to a GPI-anchored form by recombinant techniques.
用一种或多种共刺激黏附分子修饰肿瘤细胞已被提议作为开发用于人类免疫治疗的治疗性癌症疫苗的一种方法。通过基因转移使肿瘤表达B7-1（CD80）可产生一种免疫原性肿瘤细胞，该细胞可诱导抗肿瘤免疫并保护小鼠免受野生型肿瘤细胞的进一步攻击。在本报告中，我们证明将糖基磷脂酰肌醇（GPI）锚定的共刺激分子蛋白转移到肿瘤细胞膜中可作为肿瘤免疫治疗中基因转移的一种替代方法。将分离的肿瘤膜与纯化的GPI锚定的B7-1一起孵育，可在数小时内使B7-1稳定整合到肿瘤细胞膜上。用通过蛋白转移修饰以表达GPI-B7-1的EG7肿瘤膜免疫C57BL/6小鼠可诱导肿瘤特异性T细胞增殖和细胞毒性T淋巴细胞（CTL）。此外，用这些EG7膜免疫可保护小鼠免受亲代肿瘤的攻击。这里使用的蛋白转移方法不需要外源载体或活肿瘤细胞，并且在数小时内即可完成。可以使用来自新鲜或冷冻肿瘤组织的辐照细胞或膜制剂。因此，糖脂锚定分子的蛋白转移为修饰肿瘤膜用于人类免疫治疗提供了一种有效且新颖的方法。这种方法不限于共刺激分子，因为任何细胞表面蛋白都可以通过重组技术转化为GPI锚定形式。
