Sawamura Y, Hosokawa M, Kuppner M C, Kobayashi H, Aida T, Abe H, de Tribolet N
Department of Neurosurgery, Hokkaido University School of Medicine, Sapporo, Japan.
Cancer Res. 1989 Apr 1;49(7):1843-9.
The present study describes a method for in vitro expansion and characterization of antitumor-reactive lymphoid cells isolated from human malignant astrocytomas. Glioma-infiltrating lymphocytes were separated from 24 glioma specimens and cultured in medium containing interleukin 2 (50 to 2000 units/ml). Within 20 to 42 days after the initiation of culture, 20 of 24 cultures of glioma-derived lymphocytes expanded with a substantial increase in cell numbers, of at least 5 x 10(8) cells up to 5 x 10(9), with a simultaneous elimination of contaminating autologous glioma cells. The expanding glioma-derived lymphocytes consisted of 90 +/- 8% (SD) CD3+ T-cells including both CD4+ and CD8+ subpopulations. CD16 was expressed on 4 +/- 5% of the cells and three cultures studied exhibited 14% +/- 1 of Leu-19-positive cells. After 4 to 8 weeks of proliferation, interleukin 2 receptor expression decreased from 36 +/- 28% to less than 10% and the lymphocytes ceased to grow in all cultures. Glioma-derived effector lymphocytes could lyse almost all the autologous tumor targets as well as allogeneic glioma cells. The cytotoxic activity of long-term cultured peripheral blood lymphocytes obtained from the same patients appeared to be similar to that of glioma-derived lymphocytes in killing autologous tumor cells. In summary, glioma-derived lymphocytes expanded in bulk culture with high concentrations of interleukin 2 (2000 units/ml) consisted predominantly of T-lymphoblasts with the ability to kill autologous glioma cells. The tumor-infiltrating lymphocytes could be expanded to sufficient numbers for possible use in the adoptive immunotherapy of malignant gliomas.
本研究描述了一种从人类恶性星形细胞瘤中分离出的抗肿瘤反应性淋巴细胞的体外扩增及特性分析方法。从24个胶质瘤标本中分离出胶质瘤浸润淋巴细胞，并在含白细胞介素2（50至2000单位/毫升）的培养基中培养。培养开始后20至42天内，24个胶质瘤来源淋巴细胞培养物中有20个扩增，细胞数量大幅增加，至少从5×10⁸个细胞增至5×10⁹个，同时消除了污染的自体胶质瘤细胞。扩增的胶质瘤来源淋巴细胞由90±8%（标准差）的CD3⁺T细胞组成，包括CD4⁺和CD8⁺亚群。4±5%的细胞表达CD16，所研究的三个培养物显示14%±1的细胞为Leu-19阳性。增殖4至8周后，白细胞介素2受体表达从36±28%降至不足10%，所有培养物中的淋巴细胞均停止生长。胶质瘤来源的效应淋巴细胞几乎能裂解所有自体肿瘤靶细胞以及同种异体胶质瘤细胞。从同一患者获得的长期培养外周血淋巴细胞在杀伤自体肿瘤细胞方面的细胞毒性活性似乎与胶质瘤来源淋巴细胞相似。总之，在高浓度白细胞介素2（2000单位/毫升）的批量培养中扩增的胶质瘤来源淋巴细胞主要由具有杀伤自体胶质瘤细胞能力的T淋巴母细胞组成。肿瘤浸润淋巴细胞可扩增至足够数量，可能用于恶性胶质瘤的过继性免疫治疗。
