Shimizu K, Miyao Y, Okamoto Y, Matsui Y, Ushio Y, Tsuda N, Hayakawa T, Ishida N, Mogami H
No To Shinkei. 1986 Mar;38(3):265-71.
We studied whether lymphokine-activated killer (LAK) cells were capable of being induced in vitro from peripheral blood lymphocytes (PBL) of patients with malignant glioma, by using recombinant IL-2 (rIL-2). We then investigated whether they possessed anti-tumor efficacy against malignant gliomas (ONS-12, -20, -44). Human LAK cells were generated by placing 5 X 10(6) PBL into each well of 24-well plates (Corning) containing 2 ml of complete medium (CM) with 10 units of rIL-2 (TGP-3, provided by TAKEDA Chemical Industries, Ltd.). The CM consisted of RPMI 1640 with 0.1 mM nonessential amino acids, 1 microM sodium pyruvate, 5 X 10(-5) M 2-mercaptoethanol, 50 micrograms/ml gentamicin sulfate, 0.03% glutamine and 1% heat-inactivated human AB serum. The plates were incubated horizontally at 37 degrees C in a 5% CO2 atmosphere for 72-96 hours. The LAK cells were then harvested, washed three times with Hanks balanced solution, and resuspended in RPMI 1640 with 1% heat-inactivated human AB serum for the in vitro cytotoxicity assays. The anti-tumor cytotoxic activity of LAK cells was estimated in triplicate by 4-hr 51Cr release assays. The cytotoxic activity of the LAK cells against autogeneic ONS-44 glioma cells and PHA blasts was approximately 30% and a few %, respectively. The Natural Killer (NK) activity of the patient with ONS-44 glioma cells was equivalent to that of healthy subject.(ABSTRACT TRUNCATED AT 250 WORDS)
我们研究了使用重组白细胞介素-2(rIL-2)能否在体外从恶性胶质瘤患者的外周血淋巴细胞(PBL)中诱导出淋巴因子激活的杀伤(LAK)细胞。然后我们研究了这些细胞对恶性胶质瘤(ONS-12、-20、-44)是否具有抗肿瘤功效。将5×10⁶个PBL放入含有2毫升完全培养基(CM)和10单位rIL-2(由武田化学工业有限公司提供的TGP-3)的24孔板(康宁公司)的每个孔中,从而产生人LAK细胞。CM由含有0.1 mM非必需氨基酸、1 microM丙酮酸钠、5×10⁻⁵ M 2-巯基乙醇、50微克/毫升硫酸庆大霉素、0.03%谷氨酰胺和1%热灭活人AB血清的RPMI 1640组成。将平板在37℃、5%二氧化碳气氛中水平孵育72 - 96小时。然后收获LAK细胞,用汉克斯平衡液洗涤三次,并重悬于含有1%热灭活人AB血清的RPMI 1640中用于体外细胞毒性测定。通过4小时⁵¹Cr释放测定法一式三份地估计LAK细胞的抗肿瘤细胞毒性活性。LAK细胞对自体ONS-44胶质瘤细胞和成淋巴细胞的细胞毒性活性分别约为30%和百分之几。ONS-44胶质瘤细胞患者的自然杀伤(NK)活性与健康受试者相当。(摘要截短至250字)