单价抗CD20抗体诱导人B细胞淋巴瘤Raji细胞凋亡

[Apoptosis of human B cell lymphoma Raji cells induced by monovalent anti-CD20 antibody].

作者信息

Liu Yin-Xing, Fan Dong-Mei, Xiong Dong-Sheng, Xu Yuan-Fu, Shao Xiao-Feng, Xu Yuan-Sheng, Peng Hui, Yang Ming, Qin Lan, Zhu Zhen-Ping, Yang Chun-Zheng

机构信息

The National Laboratory of Experimental Hematology,Institute of Hematology,Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, PR China.

出版信息

Ai Zheng. 2003 Dec;22(12):1249-53.

PMID:14693045

Abstract

BACKGROUND & OBJECTIVE: The anti-CD20 antibodies and fragments have been applied for treatment of non-Hodgkin's lymphomas (NHL) in clinic. The new anti-CD20 antibodies and their fragments (unmodified or radiolabeled) yet have been exploited for those patients with incomplete response to rituximab. The chimeric antibody fragments Fab and F(ab) '(2) derived from HI(47)(a mouse anti-CD20 antibody) have been constructed. The present study was designed to determine the effect of HI(47) and chimeric anti-CD20 antibody fragments on growth inhibition and apoptosis of lymphoma cells.

METHODS

The binding of anti-CD20 antibodies to CD20 positive human B cell lymphoma Raji cells was examined using immunofluorescence assay. MTT method was used to evaluate the effect of chimeric antibody fragments on Raji cells growth. Annexin V staining and DNA ladder were used to examine the apoptosis of Raji cells induced by chimeric antibody fragments.

RESULTS

HI(47) and its chimeric antibody fragments were capable of binding to CD20 positive Raji cells with the binding rate of above 90%. HI(47) did not compete with rituximab in binding with Raji cells. The growth of Raji cells was inhibited by HI(47) and its fragments with the inhibition rates of (57+/-1.5)%, (65.2+/-2.5)%,and (77.2+/-3.2)%, respectively, at the concentration of 100 microg/ml. The monovalent antibody fragment Fab (20 microg/ml) induced the apoptosis of Raji cells with early apoptosis rate of 17%.

CONCLUSION

The chimeric anti-CD20 antibody fragments derived from HI(47) have inhibitory effect on Raji cells and can induce apoptosis of Raji cells.

摘要

背景与目的

抗CD20抗体及其片段已在临床上用于治疗非霍奇金淋巴瘤（NHL）。新型抗CD20抗体及其片段（未修饰的或放射性标记的）已被开发用于对利妥昔单抗反应不完全的患者。源自HI47（一种小鼠抗CD20抗体）的嵌合抗体片段Fab和F(ab)'(2)已被构建。本研究旨在确定HI47和嵌合抗CD20抗体片段对淋巴瘤细胞生长抑制和凋亡的影响。

方法

采用免疫荧光测定法检测抗CD20抗体与CD20阳性人B细胞淋巴瘤Raji细胞的结合。采用MTT法评估嵌合抗体片段对Raji细胞生长的影响。采用Annexin V染色和DNA梯状条带检测嵌合抗体片段诱导的Raji细胞凋亡。

结果

HI47及其嵌合抗体片段能够与CD20阳性的Raji细胞结合，结合率在90%以上。HI47与利妥昔单抗在与Raji细胞结合方面不存在竞争。在100μg/ml浓度下，HI47及其片段对Raji细胞的生长有抑制作用，抑制率分别为(57±1.5)%、(65.2±2.5)%和(77.2±3.2)%。单价抗体片段Fab（20μg/ml）诱导Raji细胞凋亡，早期凋亡率为17%。