干扰素α2a与白细胞介素-2联合作用对体外HepG2.2.15细胞增殖及乙型肝炎病毒复制的影响

Effect of interferon α2a combined with interleukin-2 on proliferation of HepG2.2.15 cells and replication of hepatitis B virus in vitro

目的探讨干扰素α2a(interferonα2a,IFNα2a)与白细胞介素-2(interleukin-2,IL-2)联合作用对体外肝癌细胞HepG2.2.15增殖及乙型肝炎病毒(hepatitis B virus,HBV)复制的影响。方法用不同浓度的IFNα2a(5 000、10 000、15 000 IU/ml)和IL-2(2 500、5 000、10 000 IU/ml)单独及联合作用HepG2.2.15细胞96 h后,采用WST-1法检测细胞增殖情况,倒置相差显微镜观察联合作用组细胞形态学变化,ELISA法检测联合作用组细胞上清中HBsAg和HBeAg的表达,斑点杂交法检测联合作用组细胞HBV-DNA的复制情况。结果与不同浓度的IFNα2a和IL-2单独作用相比,二者联合作用对HepG2.2.15细胞增殖的抑制作用明显增强(P<0.05);IFNα2a与IL-2联合作用组HepG2.2.15细胞显微镜下可见细胞逐渐变圆,胞浆中颗粒增多,增殖变慢,细胞间接触不紧密,细胞周围碎片增多;联合作用组细胞培养上清中HBsAg和HBeAg的表达明显受到抑制(P<0.05);5 000 IU/ml IFNα2a+10 000 IU/ml IL-2、10 000 IU/ml IFNα2a+10 000 IU/ml IL-2和15 000 IU/ml IFNα2a+10 000 IU/ml IL-2对HepG2.2.15细胞HBV-DNA的复制有抑制作用。结论 IFNα2a与IL-2联合作用对HepG 2.2.15细胞增殖及HBV-DNA复制均有抑制作用。本实验为后续体内动物实验及筛选合适的疫苗佐剂奠定了基础,也为新型乙型肝炎疫苗及治疗性乙型肝炎疫苗的研究提供了参考。

Objective To investigate the effect of interferon α2a（IFNα2a） combined with interleukin-2（IL-2） on proliferation of HepG2.2.15 cells and replication of hepatitis B virus（HBV） in vitro.Methods HepG2.2.15 cells were treated with various concentrations of IFNα2a（5 000,10 000 and 15 000 IU / ml） and IL-2（2 500,5 000 and 10 000 IU / ml） alone and in combination for 96 h respectively,the determined for proliferation by WST-1 method.The morphology of cells treated with IFNα2a in combination with IL-2 was observed by inverted phase contrast microscopy,while the expressions of HBsAg and HBeAg in supernatant by ELISA,and the replication of HBV DNA by dot blot.Results The treatment with various concentrations of IFNα2a in combination with IL-2 showed enhanced inhibitory effect on proliferation of HepG2.2.15 cells as compared with that with IFNα2a or IL-2 alone（P 0.05）.The cells treated with IFNα2a combined with IL-2 became round gradually under microscope,while the particles increased in the cytoplasm.The cells grew slowly,between which the contact was loose,and around which the pieces increased.The expressions of HBsAg and HBeAg in the culture supernatant of HepG22.2.15 cells treated with IFNα2a combined with IL-2 were inhibited significantly（P 0.05）.However,5 000 IU / ml IFNα2a ＋ 10 000 IU / ml IL-2,10 000 IU / ml IFNα2a ＋ 10 000 IU / ml IL-2 and 15 000 IU / ml IFNα2a ＋ 10 000 IU / ml IL-2 showed inhibitory effect on replication of HBV-DNA in HepG2.2.15 cells.Conclusion IFNα2a combined with IL-2 inhibited both the proliferation of HepG2.2.15 cells and the replication of HBV in vitro,which laid a foundation of further animal experiments in vivo and screening of appropriate vaccine adjuvant,and provided a reference for study on novel and therapeutic hepatitis B vaccines.