Cytotoxicity of peripheral blood lymphocyte-mediated by an anti-human ovarian carcinoma-anti-human CD3 bispecific single-chain antibody(BHL-I)in vitro and its mechanisms
10.3321/j.issn:0253-3766.2008.01.003
- VernacularTitle:抗人卵巢癌-抗人CD3单链双特异性抗体体外介导的细胞毒作用及其机制
- Author:
Jie-Zuan YANG
1
;
Lan-Juan LI
;
Jian-Qin HE
;
Xiang-Bin WANG
;
Xiao-Ning WANG
Author Information
1. 浙江大学医学院附属第一医院
- Keywords:
BHL-I;
scBsAb;
Cytotoxicity;
Cytokines
- From:
Chinese Journal of Oncology
2008;30(1):8-11
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the phenomenon that SKOV3 was killed by the PBL mediated by BHL-Iin vitro and tlle possible related mechanisms.Methods MTr colorimetric assay was used to analyze in vitro if PBL is cytotoxicitic to tlle target cells at different ratios of activated PBL and target cells (25:1,12.5:1,6.25:1,3.2:1),BHL-I mediating time(24,36,48 h),targets(SKOV3,MCF-7)and concentration of BHL-I(100,50,25,12.5,6.25,3.2,1.6,Oμg/ml).The levels of perforin,GrB mRNA and the concentrations of hIFN-γand hTNF-α,which expressed and secreted by the effector(PBL)in the process of target cell being killed,were evaluated by RT-PCR or ELISA.respectively.Results The cytotoxicity of PBL to SKOV3 cells was more efficient than to MCF-7 cells(P<0.01),and tlle PBL exhibited most effective cytotoxicity to SKOV3 cells when the concent ration ratio of PBL to SKOV3 was 12.5:1.and the mediated time of 25 μg/ml BHL-1 was 36 h.Furthermore,The PBL expressed a hisher level of perforin,GrB mRNA and hTNFF-α,hIFN-γ when the BHL-I was present,and the IL-2 increased the level of these cytokines expressed by PBL.Condusion The anti-human ovarian carcinoma×anti-human CD3 bispecific single-chain antibody BHL-I can induce PBL efficiently to kill the SKOV3 cells containing related antigens.The cytotoxic activity may be associated with the eytokines(pefforin,GrB,hTNF-α,hIFN-γ)which expressed or secreted by activated PBL,while the IL-2 Call enhance the cytotoxicity of PBL mediated by BHL-I.
