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目的探讨肺癌淋巴结细胞的培养方法及体内外杀伤作用。方法将肺癌引流淋巴结剪碎小于1 mm~3,悬于含有白细胞介素2(IL-2)的RPMI 1640培养基,置于37℃,5％CO_2培养箱培养,并行显微镜下观察和细胞计数。细胞扩增后用流式细胞仪测定细胞的免疫表型,测定其对自体肿瘤细胞的杀伤活性。将肺癌组织接种子裸鼠皮下,观察TDLN对移植瘤的抑制效果。结果淋巴组织在低浓度的IL-2培养1周,开始释放细胞群落,持续产生1～2月。1．0 g淋巴组织中在 7、14、21 d分别获得1．0×10~8、1．4×10~8和1．2×10~8细胞。表型分析显示TDLN细胞主要由CD3~+ T细胞(84．22％),和CD83~+成熟树突细胞(20．40％)组成,TDLN细胞对自体肿瘤细胞的杀伤率为 69．21％,明显低于外周血LAK细胞的杀伤率(30．17％,P<0．05)。TDLN处理后4周,移植瘤体积为(388．75±33．64)mm~3,而生理盐水组、IL-2组和LAK组分别为(742．56±160．14)mm~3、 (672．24±86．17)mm~3、(544．79±30．88)mm~3(P<0．01)。结论肺癌淋巴结细胞可在肺癌免疫治疗中发挥重要作用。
Objective To investigate the culture method of lymph node cells in lung cancer and the killing effect in vitro and in vivo. Methods Lymph node of lung cancer was cut to less than 1 mm ~ 3 and suspended in RPMI 1640 medium containing IL-2. The cells were cultured in a 5% CO 2 incubator at 37 ℃ and observed under microscope and cell count . After the cells were expanded, the immunophenotypes of the cells were measured by flow cytometry and their cytotoxic activity on the autologous tumor cells was measured. Lung cancer tissue was inoculated subcutaneously in nude mice to observe the inhibitory effect of TDLN on the xenografts. Results Lymphoid tissue was cultured in low concentration of IL-2 for 1 week to release the cell population and continued to produce from January to February. 1.0 × 10 ~ 8, 1.4 × 10 ~ 8 and 1.2 × 10 ~ 8 cells were obtained in 1.0 g lymphoid tissue at 7, 14 and 21 days respectively. Phenotypic analysis showed that TDLN cells mainly consisted of CD3 + T cells (84.22%) and CD83 + mature dendritic cells (20.40%). The killing rate of TDLN cells to autologous tumor cells was 69.21% , Significantly lower than that of peripheral blood LAK cells (30.17%, P <0.05). At 4 weeks after TDLN treatment, the volume of xenograft tumor was (388.75 ± 33.64) mm ~ 3, while it was (742.56 ± 160.14) mm ~ 3 in normal saline group, IL-2 group and LAK group, (672.24 ± 86.17) mm ~ 3, (544.79 ± 30.88) mm ~ 3 (P <0.01). Conclusion Lymph node cells of lung cancer play an important role in the immunotherapy of lung cancer.