Bast R C, Knapp R C, Donahue V C, Thurston J G, Mitchell A K, Feeney M, Schlossman S F
J Natl Cancer Inst. 1980 Feb;64(2):365-72. doi: 10.1093/jnci/64.2.365.
Antisera were raised in New Zealand White rabbits against purified populations of murine ovarian carcinoma (MOT) cells that were freed from contaminating host leukocytes and erythrocytes. In contrast to other antisera raised against this tumor, heteroantisera from rabbits immunized with purified tumor cell suspensions consistently retained antitumor activity after exhaustive absorption with syngeneic (C3HeB/FeJ) adult and fetal tissues. Absorbed antisera inhibited tumor growth in vivo and reacted with MOT cells in vitro as judged by indirect immunofluorescence, binding of staphylococcal protein A, and complement-mediated cytotoxicity. Appropriately absorbed antisera failed to bind to fetal tissues or to adult spleen, ovary, and kidney cells. Antisera with similar specificity could be obtained with the use of populations purified on a fluorescence-activated cell sorter or on discontinuous rabbit serum albumin gradients. Optimal titers against tumor were raised with multiple injections of 5 x 10(5) gradient-purified MOT cells.
用从污染的宿主白细胞和红细胞中分离出来的纯化鼠卵巢癌细胞(MOT)群体,在新西兰白兔中制备抗血清。与针对这种肿瘤制备的其他抗血清不同,用纯化肿瘤细胞悬液免疫的兔子产生的异种抗血清,在用同基因(C3HeB/FeJ)成年和胎儿组织进行彻底吸收后,始终保留抗肿瘤活性。通过间接免疫荧光、葡萄球菌蛋白A结合以及补体介导的细胞毒性判断,吸收后的抗血清在体内抑制肿瘤生长,在体外与MOT细胞发生反应。适当吸收的抗血清不能与胎儿组织或成年脾脏、卵巢和肾脏细胞结合。使用在荧光激活细胞分选仪或不连续兔血清白蛋白梯度上纯化的群体,可以获得具有相似特异性的抗血清。通过多次注射5×10⁵个梯度纯化的MOT细胞,可产生针对肿瘤的最佳效价。