Eisenthal A, Rosenberg S A
Surgery Branch, National Cancer Institute, Bethesda, MD 20892.
Clin Exp Metastasis. 1988 Sep-Oct;6(5):387-400. doi: 10.1007/BF01760574.
We have cross-linked, using succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) as a heterobifunctional reagent, anti-B16 melanoma monoclonal antibody to lymphokine activated killer (LAK) cells, independent of the Fc receptor. The conditions of such linkage were optimized so that the cytotoxic properties of LAK cells, as measured in a 4 h chromium release assay against fresh tumor cells, were preserved. Using the techniques described here, covalent cross linking of anti-B16 antibody to LAK cells preserved the reactivity of this antibody to antigens on B16 melanoma cells, and preserved the cytotoxic properties of the antibody-bound LAK cells to lyse B16 tumor cells and other tumor cells in vitro. Cross-linking antibody remained active on the surface of LAK cells for as long as 24 h after the completion of binding. Treatment of established B16 melanoma pulmonary or subcutaneous (s.c.) tumors with LAK cells cross-linked to anti-B16 melanoma monoclonal antibody did not significantly alter their therapeutic efficacy over untreated cells. The possible explanations for these in vivo observations and suggested approaches to increase the efficacy of the cross-linked LAK cells are discussed.
我们使用琥珀酰亚胺基4-(N-马来酰亚胺甲基)环己烷-1-羧酸酯(SMCC)作为异双功能试剂，将抗B16黑色素瘤单克隆抗体与淋巴因子激活的杀伤(LAK)细胞交联，该交联不依赖于Fc受体。对这种交联的条件进行了优化，以便在针对新鲜肿瘤细胞的4小时铬释放试验中测定的LAK细胞的细胞毒性特性得以保留。使用本文所述的技术，抗B16抗体与LAK细胞的共价交联保留了该抗体对B16黑色素瘤细胞上抗原的反应性，并保留了抗体结合的LAK细胞在体外裂解B16肿瘤细胞和其他肿瘤细胞的细胞毒性特性。交联抗体在结合完成后在LAK细胞表面保持活性长达24小时。用与抗B16黑色素瘤单克隆抗体交联的LAK细胞治疗已建立的B16黑色素瘤肺或皮下(s.c.)肿瘤，与未处理的细胞相比，其治疗效果没有显著改变。讨论了这些体内观察结果的可能解释以及提高交联LAK细胞疗效的建议方法。
