重组IFN-α-IL-18对鸡淋巴细胞组胺的诱导生成和NF-κB p65的激活  

Recombinant IFN-α-IL-18 increases level of histamine produced by chicken lymphocyte and activate NF-κB p65 activity

作　　者：杨丹芳[1] 李银聚[1] 杨惠[1] 刘一尘[1] 程相朝[1] 张春杰[1] 

机构地区：[1]河南科技大学动物疫病与公共卫生重点实验室,洛阳471003

出　　处：《中国免疫学杂志》2017年第7期995-999,共5页Chinese Journal of Immunology

基　　金：河南省科技攻关项目(132102110117)

摘　　要：目的:探讨鸡IFN-α-IL-18融合蛋白对鸡外周血淋巴细胞组胺诱导产生和NF-κB p65的激活与转运。方法:无菌采取健康鸡抗凝血,分离鸡外周血淋巴细胞,以RPMI1640完全培养液调整细胞密度为5×10~6细胞/ml,将细胞分成10组,分别加入酵母表达并纯化的IFN-α-IL-18,IL-18和IFN-α蛋白终浓度250 ng/ml,500 ng/ml和1 000 ng/ml,对照组细胞仅加RPMIl640培养基,每组各设3个重复,37℃、5%CO_2培养箱培养48 h,分别测定淋巴细胞中组氨酸脱羧酶活性,组胺含量,IFN-γ,PI3K,MAPK和细胞核中NF-κB p65的含量。结果:检测结果表明,重组IFN-α-IL-18和IL-18均能显著促进鸡外周血淋巴细胞中组氨酸脱羧酶活性(P<0.01),增加组胺含量(P<0.01),诱生IFN-γ(P<0.01),提高PI3K(P<0.01)和细胞核中NF-κB p65的含量(P<0.01),较高浓度的IFN-α对淋巴细胞有类似的作用;IFN-α-IL-18、IL-18和IFN-α对鸡淋巴细胞中的MAPK作用不明显,仅在较高浓度下IFN-α-IL-18和IL-18对细胞中MAPK产生影响(P<0.05)。结论:IFN-α-IL-18及IL-18能够明显促进淋巴细胞中组胺的生成,并由此加强IFN-γ的诱生;IFN-α-IL-18、IL-18和IFN-α可能通过促进淋巴细胞中PI3K的表达激活NF-κB并促进NF-κB p65的核质转运,这为进一步开展IFN-α-IL-18融合蛋白的功能研究及其在疫病防控中的作用机制探索奠定了基础。Objective： To discuss the effects of recombinant IFN-α-IL-18 fusion protein on chicken lymphocyte histamine induced and the NF-κB p65 activation and nucleo-cytoplasmic transport. Methods： The healthy chickens blood was sterile adopted with anticoagulant,then separation of the chicken peripheral blood lymphocyte and divided into 10 groups： The yeast expression and purification protein IFN-α-IL-18, IL-18, IFN-α were added with 250 ng/ml,500 ng/ml and 1 000 ng/ml respectively while the control was only added RPMI1640 with 3 repetitions for each group. Then the histidine decarboxylase activity, histamine, IFN-γ, PI3K, MAPK and NF-κB p65 in cell nucleus were detected. Results： The recombinant IFN-α-IL-18 and IL-18 could significantly promote the activity of histidine decarboxylase （P〈0. 01 ） ,increase the contents of histamine （P〈0. 01 ） ,induce IFN-γ （P〈0. 01 ） ,improve the contents of PI3K （P〈0. 01） and the NF-κB 1365 levels in nucleus （P〈0. 01） ,and the higher concentration of IFN-α had a similar effect to lymphocytes. The effects of IFN-α-IL-18, IL-18 and IFN-α on MAPK was acratia. Conclusion： IFN-α-IL-18 and IL-18 can stimulate chicken peripheral blood lymphocyte populations increased histamine contents significantly and promote the induction of IFN-α. IFN-α- IL-18,IL-18 and IFN-α increase PI3K expression in lymphocyte associated with the NF-KB activation and NF-κB 1o65 nucleo- cytoplasmic transport. The study built foundation for the function of IFN-α-IL-18 and the exploration of the mechanism of controlling epidemic diseases.

关 键 词：IFN-α-IL-18 组氨酸脱羧酶 组胺 PI3K 核因子ΚB 

分 类 号：S852.4[农业科学—基础兽医学]