双特异抗体导向骨髓中活化T细胞清除骨髓中肿瘤细胞

Targeting T Lymphocytes by Bi-specific Monoclonal Antibodies to Eliminate Tumor Cells in Bone Marrow

先用抗CD3单抗和IL-2激活肺癌患者骨髓中的T淋巴细胞,然后按不同比例将人肺癌细胞株PC84045细胞掺入到骨髓细胞中,再加入能同时识别CD3抗原和人肺癌细胞表面抗原的双特异抗体CD3-ALTO4,在IL-2和IL-3存在下共同培养3d后测试.在T细胞与PC84045癌细胞之比为8:1时,对PC84045肺癌细胞净化效率为4Logs,16;1时为5Logs,并在NC裸鼠体内证实清除效果是确切的.净化后的骨髓CFU-GM、BFU-E收获率均>85%(P>0.05;P>0.05);而且净化后的骨髓中的T细胞仍保持着对PC84045肺癌细胞很强的杀伤能力(P<0.01).双特异抗体CD3-ALTO4能有效地导向骨髓中活化T细胞清除污染的肿瘤细胞,造血干细胞无明显损伤.本文方法处理的骨髓细胞还保持着高度特异的细胞毒活性,移植这样的骨髓细胞将会在体内进一步清除化疗残存的肿瘤细胞,加速移植后免疫功能再建,降低复发.

PC84045 human lung carcinoma cells (PC84045 HLCCs) were mixed with the BM cells activated by anti-CD3 McAb and IL-2 and then CD3-ALTO4 bi-specific McAbs was put into the cell suspension. The mixed cells were cultured for 3 days in presence of IL-2 and IL-3. Tumor cell colony formation method in half-solid culture system was used to assess the purging efficiency (PE) of the activated T cells to PC84045 HLCCs. The ratio of T cells to PC84045 HLCCs was 8 to 1, the PE was 4 Logs. The rario of T cells to PC84045 HLCCs was 16 tol, the PE was 5 Logs. When the purged BM cells were injected into the NC nude mice, none of subcutaneous node was found. The number of the CFU-GM, BFU-E of the purged BM cells was more than 85 % compared with that of the fresh BM cells (P<0.05, P<0.05). Also, it was significant that purged BM cells had specifically got the high cytotoxicity to PC84045 HLCCs P<0.01). CD3-ALT04 bi-specific McAb can target the activated T cells in BM cells to eliminate the tumor cells by an specific recognization to cell surface antigen. This method does not damage the heamotopoietic stem cells, and, further, induce the specific tumoricidial function of T cells from BM. To transplant those activated BM cells may eradicate the residual tumor cells in vivo after high-dose chemotherapy and accelerate the recovery of immune function to reduce the relapse of disease.