[In vitro cytolysis of B-lymphoma cells mediated by an anti-CD3/anti-CD20 bispecific single-chain antibody].
After having successfully constructed and expressed the gene of the anti-CD3/anti-CD20 bispecific single-chain antibody (bscCD3 x CD20), here we analyzed its in vitro bioactivity of mediating the lysis of Ramous human B-lymphoma cells in the presence of T-enriched human peripheral blood lymphocytes (PBL). Obvious opoptosis characters were observed by Annexin V/PI(AV/PI) stained and scanning electron microscope. As evaluated by non-radioactive cytotoxity assay, the bscCD3 x CD20 showed potent bioactivity of mediating human B-lymphoma cells lysis in the presence of T-enriched human PBL. The potency of cytotoxicity depended on the ratios of effect cells to target cells (E:T) used. Further, the antibody showed a dose and time-dependent effect on mediating Ramous cells lysis. The specific lysis reached about 87.3% at an antibody concentration of 5microg/mL and E:T used at 10:1. Clear changes in apoptogenes expression profiles were detected by apoptosis gene array after Ramous cells were treated with the antibody and PBL. Among the upregulated apoptogenes, ATM and P53 showed an increase of 187 times and 15 times respectively, which suggested that ATM-p53 pathway may be the main apoptosis way of Ramous cells induced by T cells in the presence of the bscCD3 x CD20.
Yu
Rui
R
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Li
Shi-Chong
SC
Wu
Ben-Chuan
BC
Liu
Hong
H
Ye
Ling-Ling
LL
Liu
Xing-Mao
XM
Wang
Qi-Wei
QW
Chen
Zhao-Lie
ZL
D018033:Antibodies, Bispecific / Q000276:immunology*; D018951:Antigens, CD20 / Q000276:immunology*; D017209:Apoptosis / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000276:immunology* / Q000473:pathology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014407:Tumor Cells, Cultured
D018033:Antibodies, Bispecific; D018951:Antigens, CD20; D017252:CD3 Complex