小鼠CTL体外非特异性扩增对其杀瘤活性的影响

目的 在体外用抗CD3单抗、抗CD28单抗和白细胞介素2（IL－2）扩增肿瘤特异性CTL，为肿瘤过继治疗提供足够数量的、具有高度杀瘤活性的效应细胞。方法 采用两种方案培养肿瘤细胞免疫的小鼠脾细胞。（1）抗CD3单抗刺激48h后，加入抗CD3单抗和20U／mlrIL－2（抗－CD3＋IL－2组）；（2）抗CD3单抗和抗CD28单抗同时刺激48h后，加入抗CD3单抗、抗CD28单抗和20U／ml rIL－2（抗－CD3＋抗－CD2＋IL－2组）。分别检测2组效应细胞的增殖水平、杀瘤活性及表型。结果 抗CD3＋IL－2组细胞的^3H－TdR掺入量在第6天、12天、20天分别为22126、52426、2072，抗－CD3＋抗－CD28＋IL－2组细胞的^3H-TdR掺入量在第6天、12天、30天分别为32168、12922、3265，后者明显高于前者。在培养12d时，抗－CD3＋IL－2组细胞对FBL03的最大杀伤活性为66．4％，抗－CD3＋抗－CD28＋IL－2组细胞对FBL－3的最大杀伤活性为77．8％。细胞表型FACS分析结果表明，抗－CD3＋抗－CD28＋IL－2组培养12d后的细胞90％以上为Thy1.2^ 细胞，且CD25^ 细胞在抗－CD3＋抗－CD28＋IL－2组、抗－CD3＋IL－2组分别为23．00％、8．15％。结论 抗CD3单抗、抗CD28单抗和低剂量IL－2同时非特异性刺激，可获得大量扩增的、具有高度杀瘤活性的肿瘤特异性CTL。

Anti-tumor effects of mouse CTL of non-specific expansion

Objective To obtain maximal expansion and anti tumor activity of CTL for tumor adoptive immunotherapy. Methods Anti CD3 McAb, anti CD28 McAb and rIL 2 were employed to activate and expand CTL. Splenocytes from tumor immunized mice were activated in vitro by either anti CD3 McAb for 48 hours and then cultured in anti CD3 McAb plus 20U/ml rIL 2 (the anti CD3 IL 2 group) or anti CD3 McAb, anti CD28 McAb plus 20U/ml rIL 2 (the anti CD3 anti CD28 IL 2 group). The proliferation, anti tumor activity of tumor specific T cells and cell phenotypes were studied in both groups. Results 3H TdR incorporation (cpm) in the anti CD3 IL 2 group were 22126, 5426 and 2072 on day 6, 12 and 20 respectively, while 3H TdR incorporation (cpm) in the anti CD3 anti CD28 IL 2 group were 32168, 12922 and 3265 on day 6, 12 and 20 respectively. The maximum anti tumor activities of both groups were 66.4% and 77.8% respectively on day 12. More than 90% effective cells in the anti CD3 anti CD28 IL 2 group were Thy1.2 on day 12. The percentage of CD25 cells in the anti CD3 anti CD28 IL 2 group and the anti CD3 IL 2 group were 23.00%, 8.15% respectively. Conclusion The protocol for obtaining remarkable proliferation and anti tumor activity of tumor specific CTL has been successfully optimized.