Kikuchi T, Sakai H, Nakamura N, Watanabe M, Ohno T
Department of Neurosurgery, Tokyo Jikei University School of Medicine, Japan.
No To Shinkei. 1990 Jun;42(6):575-80.
We analyzed cytolytic activity and cell surface phenotypes of LAK (lymphokine activated killer) cells which were stimulated with recombinant IL-2 and an anti-CD3 antibody. This study involved 9 patients with astrocytomas, one patient with ganglioglioma, and one patient with medulloblastoma and their ages ranged between 5 and 80yr. Peripheral blood lymphocytes were separated from about 40 ml venous blood on a Ficoll-Paque gradient. After PBL's were cultured with r-IL 2 and anti-CD 3 antibody for about two weeks, more than 10(8) LAK cells could be obtained. Their cytolytic activity was measured by standard 4 hours-51Cr release assay and K 562, Daudi, U 251 MG, MCF-7, and autologous tumor cells were used as target cells. Their surface phenotypes (CD 3, CD 4, CD 8, CD 16, CD 25, Leu 7, HLA-DR) were measured by FACS. These LAK cells showed weaker killing activity against all kinds of tumor cells than usual LAK cells (LAK cells stimulated with r-IL 2 alone) did. Their surface phenotypes were more sensitive to CD 3 and less sensitive to CD 16 and Leu 7 than those of usual LAK cells. One of weak points of so-called LAK therapy is that many PBL's have to be obtained by leukapheresis and because of that, it is difficult to undertake LAK therapy to children or elderly patients. However, by using an anti-CD 3 antibody, enough LAK cells as effector of LAK therapy could be obtained more easily.(ABSTRACT TRUNCATED AT 250 WORDS)
我们分析了用重组白细胞介素-2和抗CD3抗体刺激的淋巴因子激活的杀伤细胞(LAK细胞)的细胞溶解活性和细胞表面表型。本研究纳入了9例星形细胞瘤患者、1例神经节胶质瘤患者和1例髓母细胞瘤患者,年龄在5至80岁之间。通过Ficoll-Paque梯度从约40ml静脉血中分离外周血淋巴细胞。外周血淋巴细胞与重组白细胞介素-2和抗CD3抗体培养约两周后,可获得超过10⁸个LAK细胞。通过标准的4小时⁵¹Cr释放试验测定其细胞溶解活性,并使用K562、Daudi、U251MG、MCF-7和自体肿瘤细胞作为靶细胞。通过荧光激活细胞分选术(FACS)测定其表面表型(CD3、CD4、CD8、CD16、CD25、Leu7、HLA-DR)。这些LAK细胞对各种肿瘤细胞的杀伤活性比普通LAK细胞(仅用重组白细胞介素-2刺激的LAK细胞)弱。与普通LAK细胞相比,它们的表面表型对CD3更敏感,对CD16和Leu7不太敏感。所谓LAK疗法的一个缺点是必须通过白细胞分离术获取大量外周血淋巴细胞,因此,对儿童或老年患者进行LAK疗法很困难。然而,通过使用抗CD3抗体,可以更轻松地获得足够数量的LAK细胞作为LAK疗法的效应细胞。(摘要截短于250字)