Expression of the immune inhibitors of human B cell activating factor belonging to the tumor necrosis factor family and evaluation of immune response in vaccinated mice
- VernacularTitle:肿瘤坏死因子家族的B细胞激活因子免疫抑制分子的表达及免疫原性分析
- Author:
Hui-Guang GAO
;
Feng-Tian HE
;
Rong-Fen LI
;
Qing JI
;
Gang HUANG
;
Da-Qiang HU
;
Li ZHANG
;
Wei GONG
;
Ying HU
- Publication Type:Journal Article
- Keywords:
Tumor necrosis factors;
Suppressor factor;
immunologic;
T-help cell epitope
- From:
Chinese Journal of Rheumatology
2001;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare T-helper cell(Th)epitnpe-modified human soluble B cell activating factor belonging to the tumor necrosis factor family(BAFF)mutants and evaluate their immune response in vaccinated mice.Methods Recombinant cDNAs were cloned and ligated into the prokaryotie expression vec- tor pQE-80L respectively.The recombinant proteins were induced to express by IPTG in E.coli DH5?and purified with Ni-NTA chromatography.BALB/c mice were immunized with recombinant proteins respectively and the titres of the antibodies that were cross-reactive with BAFF in sera were analyzed by ELISA.Inhibiting ability of the antibodies in sera was analyzed by MTT assay.Results The recombinant proteins were highly expressed in E.coli DH5?.After purification,the purity of recombinant proteins was more than 90%.BALB/c mice immunized with recombinant proteins produced high levels of BAFF-specific antibodies.Cell proliferation assay showed that the sera of immunized mice could inhibit the proliferation-inducing activity of recombinant sBAFF and natural sBAFF.Conclusion The immune inhibitors of human BAFF which can induce polyclonal antibodies that are cross-reactive with BAFF are successfully prepared.These results may provide the basis for further study of their therapeutic effects.
