Okamoto Y, Shimizu K, Miyao Y, Ushio Y, Matsui Y, Hayakawa T, Tsuda N, Mogami H
No To Shinkei. 1986 Mar;38(3):233-7.
We have studied the in vitro antitumor effectiveness of murine lymphokine-activated killer (LAK) cells induced by recombinant IL-2 (rIL-2). LAK cells were generated by placing 5 X 10(7) fresh C 57 BL/6 splenocytes (erythrocytes were lysed osmotically) in 10-cm (diameter) dishes (Falcon) containing 10 ml of complete medium (CM). The CM consisted of RPMI 1640 with 0.1 mM non-essential amino acids, 1 microM sodium pyruvate, 5 X 10(-5)M 2-mercaptoethanol, 50 micrograms/ml gentamicin sulfate, 0.03% glutamine, 10% heat-inactivated fetal calf serum (FCS) and 10 units/ml of rIL-2 (TGP-3, provided by TAKEDA Chemical Industries, Ltd). The dishes were incubated horizontally at 37 degrees C in a 5% CO2 atmosphere for 72-96 hr. The LAK cells were then harvested, washed three times, and resuspended in RPMI 1640 with 5% heat-inactivated FCS for the in vitro cytotoxicity assay. The antitumor cytotoxic activity of LAK cells was estimated in triplicate by 4 hr 51Cr release assays. The cytotoxic activity of LAK cells against syngeneic 203 glioma and normal syngeneic glioblasts was approximately 50% and a few %, respectively. The in vitro cytotoxicity of LAK cells against syngeneic EL-4 thymoma, allogeneic YAC-1 lymphoma and P-815 mastocytoma was 72%, 87% and 43%, respectively. Thus LAK cells have apparent tumor specificity in vitro and are easily generated. Fresh splenocytes of CBA/J mice were markedly lytic for natural killer (NK)-sensitive YAC-1 cells, but not for 203-glioma cells or NK-resistant P-815 cells.(ABSTRACT TRUNCATED AT 250 WORDS)
我们研究了重组白细胞介素-2(rIL-2)诱导的小鼠淋巴因子激活的杀伤(LAK)细胞的体外抗肿瘤效果。通过将5×10⁷个新鲜的C57BL/6脾细胞(红细胞经渗透压裂解)置于含有10ml完全培养基(CM)的10厘米(直径)培养皿(Falcon)中,来生成LAK细胞。CM由添加了0.1mM非必需氨基酸、1μM丙酮酸钠、5×10⁻⁵M 2-巯基乙醇、50μg/ml硫酸庆大霉素、0.03%谷氨酰胺、10%热灭活胎牛血清(FCS)和10单位/ml rIL-2(TGP-3,由武田化学工业株式会社提供)的RPMI 1640组成。将培养皿在37℃、5%二氧化碳气氛中水平孵育72 - 96小时。然后收获LAK细胞,洗涤三次,并重悬于含有5%热灭活FCS的RPMI 1640中用于体外细胞毒性测定。通过4小时的⁵¹Cr释放试验一式三份地估计LAK细胞的抗肿瘤细胞毒性活性。LAK细胞对同基因203胶质瘤和正常同基因胶质母细胞的细胞毒性活性分别约为50%和百分之几。LAK细胞对同基因EL-4胸腺瘤、异基因YAC-1淋巴瘤和P-815肥大细胞瘤的体外细胞毒性分别为72%、87%和43%。因此,LAK细胞在体外具有明显的肿瘤特异性且易于生成。CBA/J小鼠的新鲜脾细胞对自然杀伤(NK)敏感的YAC-1细胞有明显的裂解作用,但对203胶质瘤细胞或NK抗性的P-815细胞没有作用。(摘要截短于250字)