协同刺激信号α-CD28单抗增强肿瘤特异性CTL体内外的杀瘤活性

Anti-CD28 McAb Enhance Anti tumor Activity of Tumor Specific Cytotoxic T Lymphocyte in Vitro and in Vivo

目的 采用α CD2 8单抗增强α CD3单抗刺激的肿瘤特异性T细胞的杀瘤活性。方法 采用 2种方案培养FBL 3免疫的小鼠脾细胞。 (1)使用α CD3单抗 48h后 ,加入α CD3单抗和 2 0U/mlrIL 2 (α CD3 +IL 2组 ) ;(2 )α CD3单抗和α CD2 8单抗同时使用 48h后 ,加入α CD3单抗、α CD2 8单抗和 2 0U/mlrIL 2 (α CD3+α CD2 8+IL 2组 )。3H TdR掺入法检测两组效应细胞的增殖水平。标准 4h 51Cr释放法检测两组效应细胞的杀伤活性。在培养第 12天 ,每只荷瘤小鼠过继转输 1× 10 7CTL ,观察小鼠的存活期。结果 α CD3 +IL 2组、α CD3+α CD2 8+IL 2组细胞的3H TdR掺入量分别为 8.36× 10 4 、1.31× 10 5,后者明显高于前者。在培养 12d时 ,效靶比为 12 .5∶1时 ,α CD3+IL 2组细胞对FBL 3的特异性杀伤活性为 35 .6 % ,α CD3 +α CD2 8+IL 2组细胞对FBL 3的特异性杀伤活性为 49.5 %。分别过继转输两组效应细胞治疗荷瘤小鼠 ,荷瘤小鼠的平均存活期分别为 16 .7和 2 0 .8d。结论 协同刺激信号α CD2

Purpose To enhance anti tumor activity of α?CD3 McAb activated tumor specific cytotoxic T lymphocyte(CTL) by α?CD28 McAb in vitro and vivo for tumor adoptive immunotherapy. Methods Two different modles were applied to culture splenocytes from FBL?3 immunized mice:(1) activated by α?CD3 McAb for 48 h and then cultured in α?CD3 McAb plus 20 U/ml rIL?2(α?CD3+IL?2 group);(2) activated by α?CD3 McAb,α?CD28 McAb for 48 h and then cultured in α?CD3 McAb,α?CD28 McAb plus 20 U/ml rIL?2(α?CD3+α?CD28+IL?2 group).The proliferation ( 3H?TdR incorporation assay)and anti tumor activity(standard 4 h? 51 Cr release assay) of FBL?3 specific CTL were studied in both groups.On day 12,each bearing tumor mouse was transferred with 1×10 7 CTL by ip. Results 3H?TdR incorporation (cpm) in α?CD3+IL?2 group and α?CD3+α?CD28+IL?2 group were 8.36×10 4,1.31×10 5 respectively.On day 12,the anti tumor activities of both groups were 35.6% and 49.5% respectively (E/T=12.5/1).The CTL of α?CD3+IL?2 group and α?CD3+α?CD28+IL?2 group were transferred to bearing tumor mouse respectively.The lifespans of bearing tumor mouse were 16.7 d and 20.8 d respectively. Conclusions α?CD28 McAb can enhance antitumor activity of tumor specific CTL activated by α?CD3 McAb.