肺癌淋巴结细胞培养及体内外的杀伤作用

Study on the culture of the tumor-draining lymph node tissue of lung cancer patients and its killing and wonding activity both in vivo and in vitro

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 of the tumor-draining lymph node tissue （TDLT） of lung cancer patients and its killing and wonding activity both in vivo and in vitro. Methods The tumor-draining lymph node tissue （TDLT） of lung cancer patients was cultured with low dose IL-2 and be counted. Cell growth, cell surface markers and its killing and wounding activity were determined. Tumor was implanted preventively into subcultaneous tissues of nude mice. The TDLN was used on treating the implanted tumor and the therapeutic effect was observed. Results We generated 1.0,1.4 and 1.2 × 10^8 cells when 1.0 g TDLT was cultured 7,14,21 days. The majority of the cells were CD3-posirive T cells （84.22%） and CD83-positive ceils （20.40%）. The ratio of TDLN killing the tumor cells （69.21% ） is lower than the ratio of LAK cells （30.17 %, P 〈 0.05）. TDLN coudld inhibit the growth of the implanted tumor. The average sizes of TDLN groups, IL-2 groups and LAK groups were （388.75 ±33.64）, （742.56± 160.14）, （672.24 ± 86.17）, （544.79± 30.88） mm3 （ P 〈 0.01 ）. Conlusion TDLT may be play an important role while used for immunotherapy for patients with lung cancer.