Effects of recombinant IFN-α-IL-18 on chicken lymphocyte transformation and NF-κB activity in vitro
10.3969/j.issn.1000-484X.2015.08.008
- VernacularTitle:重组IFN-α-IL-18体外对鸡外周血淋巴细胞转化及核因子κB的影响
- Author:
Hui YANG
;
Yinju LI
;
Yichen LIU
;
Xiangchao CHENG
;
Danfang YANG
;
Haifeng HAN
;
Xiuzhe JIN
- Publication Type:Journal Article
- Keywords:
Interferon-α-interleukin-18;
Yeast expression;
The lymphocyte transformation test;
NF-κB
- From:
Chinese Journal of Immunology
2015;(8):1040-1044,1048
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To discuss the effects of recombinant IFN-α-IL-18 fusion protein on chicken lymphocyte transformation and the chicken nuclear factor kappa B ( NF-κB ) activity.Methods: The recombinant plasmids pPICZ-IFN-α, pPICZ-IL-18 and pPICZ-IFN-α-IL-18 were constructed,and transformed into P.Pastoris X-33 strain by electroporation.The recombinant proteins were ex-pressed under the induction of 1% methanol,and detected by SDS-PAGE and Western blot.The biological activities of the expressed products were detected by the lymphocyte transformation test,the NF-κB concentration in chicken lymphocyte and lymphocytic nucleus were detected at different times with ELISA.Results:SDS-PAGE and Western blot showed that the expressed products existed in super-natant,and the molecular weights were about 22 kD,23 kD and 43 kD,respectively.The expressed products IL-18 and IFN-α-IL-18 could stimulate chicken lymphocyte transformation (P<0.05),the biological activity of IFN-αstimulating lymphocyte transform was feeble (P>0.05).Compared with the control group,the total NF-κB concentration in chicken lymphocyte induced by IL-18 and IFN-α-IL-18 were increased (P<0.05),and the NF-κB in lymphocytic nucleus was increased significantly (P<0.01).Otherwise,the total NF-κB in lymphocyte induced by IFN-αincrease was limited (P>0.05),but the NF-κB in lymphocytic nucleus showed the remarkable increase ( P<0.05 ) .Conclusion: IFN-α-IL-18 and IL-18 can promote lymphocyte transformation significantly, the activity of IFN-αinduced lymphocyte transformation is imperfect.The biological activity of stimulating lymphocyte transformation is associated with the NF-κB expression,activation and nucleo-cytoplasmic transport.The study built foundation for the function of IFN-α-IL-18 fusion protein and the exploration of the role of controlling epidemic diseases.
