抗CD_3/抗CD_(20)偶联物的制备及其介导激活T细胞杀伤活性研究

Preparation of anti CD_3/anti CD_(20) Heteroconjugate and Its Specific T Cell Mediated Lysis

目的:研究抗CD3/抗CD20抗体偶联物介导的激活T细胞杀伤活性。方法:通过SPDP偶联,经分子筛层析法纯化得到抗CD3/抗CD20抗体偶联物,并用FACS和玫瑰花环试验鉴定纯化产物;采用51Cr释放试验测定该抗体偶联物介导的体外靶向杀伤活性。结果:纯化的抗CD3/抗CD20抗体偶联物具有与Jurkat(CD3+)和Daudi细胞(CD20+)的结合活性,且能同时将Jurkat和Daudi细胞结合形成玫瑰花环;在体外能介导激活的T细胞杀伤Daudi细胞。结论:抗CD3/抗CD20抗体偶联物体外能介导激活的T细胞杀伤表达CD20抗原的肿瘤细胞,为B细胞恶性肿瘤临床治疗提供实验依据。

Objective:To study the T cell-mediated lysis of malignant human B cells by an ti-CD 3 /anti-CD 20 heteroconjugate.Methods:The heteroconjugate was prepared by SPDP meth ods and purified by size exclusion chromatography.The product was identified by FACS and rosette formation assay.The effect of heteroconjugate mediated lysis of CD 20 -expressing tumor cells was determined by 51 Cr release assay in vitro.Results:The anti-CD 3 /anti-CD 20 hetero con jugate could specifically bind both Jurkat cells(CD 3 +)and Daudi cells(CD 20 +)and cross-link these target cells.The heteroconjugate appeared to be potent in targeting activated peripheral blood mononu-clear cell(PBMC)to lyse Daudi cells in vitro.Conclusion:Our study showed an im portant role of bispecific antibody for retargeting peripheral blood lymphocytes to lyse CD 20 antigen expressing tumor cells.