小鼠CTL体外非特异性扩增对其杀瘤活性的影响  

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

作　　者：黄辉[1] 俞红[2] 熊思东[1] 林云璐[1] 

机构地区：[1]复旦大学医学院免疫学教研室,上海200032 [2]复旦大学医学院微生物学教研室,上海200032

出　　处：《中华微生物学和免疫学杂志》2001年第2期143-145,共3页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目! ( 3 95 70 80 8;3 93 70 773 )

摘　　要：目的　在体外用抗CD3单抗、抗CD2 8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 2 0U mlrIL 2 (抗 CD3+IL 2组 ) ;(2 )抗CD3单抗和抗CD2 8单抗同时刺激 48h后 ,加入抗CD3单抗、抗CD2 8单抗和 2 0U mlrIL 2 (抗 CD3+抗 CD2 8+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、2 0天分别为 2 2 12 6、5 42 6、2 0 72 ,抗 CD3+抗 CD2 8+IL 2组细胞的3H TdR掺入量在第 6天、12天、2 0天分别为 32 16 8、12 92 2、32 6 5 ,后者明显高于前者。在培养 12d时 ,抗 CD3+IL 2组细胞对FBL 3的最大杀伤活性为 6 6 .4% ,抗 CD3+抗 CD2 8+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 CD2 8+IL 2组培养 12d后的细胞 90 %以上为Thy1.2 + 细胞 ,且CD2 5 + 细胞在抗 CD3+抗 CD2 8+IL 2组、抗 CD3+IL 2组分别为 2 3.0 0 %、8.15 %。结论　抗CD3单抗、抗CD2 8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。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.

关 键 词：抗CD28单抗 抗CD3单抗 肿瘤特异性T细胞 肿瘤过继疗法 

分 类 号：R730.5[医药卫生—肿瘤]