Shao Shengwen, Zhou Hongchang, Tong Yimin, Ren Yanli, Chen Zhihui
Department of Microbiology, Medical School of Huzhou Teachers College, Huzhou 313000, China.
Wei Sheng Yan Jiu. 2011 May;40(3):295-8.
To explore the feasibility of induction of neutralization antibodies against hepatitis C virus (HCV) infection by HCV envelope 2 protein (E2) DNA vaccines immunization.
Two kinds of expression plasmids of HCV envelope 2 protein, plasmid pCI-1b661 Delta encoding hydrophobic carboxyl terminal truncated E2 and pCI-1b661 Delta encoding E2 with deletion of hypervariable region 1 (HVR1) and carboxyl terminal, were constructed and respectively transfeted 293T cells, and truncated E2 protein in whole cell lysate and supernatant of 293T cells were analyzed by Western blot. After BALB/c mouse were intramuscularly immunized by the plasmids, sera antibodies against HVR1 were detected by ELISA and the neutralization activity of the antibodies were assayed with HCV pseudotype particle (HCVpp).
Both plasmids could express secretary truncated E2 protein. All the mice immunized with plasmid pCI-1b661 produced HVR1 antibodies,while no HVR1 antibodies were detected in pCI-1b661 Delta immunized mice. The sera neutralization percentages against HCVpp in pCI1lb661 Delta and pCI-lb661 Delta immunized mice were (78.5 +/- 13.8)% and (38.7 +/- 6.5)%, respectively (P <0.01). Sera neutralization activity against HCVpp was positive correlated with the level of HVR1 antibodies in pCI-1b661 immunized mice (r = 0.967, P<0.01).
DNA vaccines expressing truncated E2 protein could induce neutralization antibodies against HCV, and neutralization antibodies mainly was consisted of the antibodies against HVR1.
探讨丙型肝炎病毒(HCV)包膜2蛋白(E2)DNA疫苗免疫诱导抗HCV感染中和抗体的可行性。
构建两种HCV包膜2蛋白表达质粒,即编码疏水羧基末端截短E2的质粒pCI-1b661 Delta和编码缺失高变区1(HVR1)及羧基末端的E2的质粒pCI-1b661 Delta,分别转染293T细胞,用蛋白质免疫印迹法分析293T细胞全细胞裂解物和上清液中的截短E2蛋白。用这些质粒对BALB/c小鼠进行肌肉免疫后,用酶联免疫吸附测定法检测血清中抗HVR1抗体,并用HCV假型颗粒(HCVpp)测定抗体的中和活性。
两种质粒均可表达分泌型截短E2蛋白。用质粒pCI-1b661免疫的所有小鼠均产生了HVR1抗体,而用pCI-1b661 Delta免疫的小鼠未检测到HVR1抗体。pCI1lb661 Delta和pCI-lb661 Delta免疫小鼠血清对HCVpp的中和百分比分别为(78.5±13.8)%和(38.7±6.5)%(P<0.01)。pCI-1b661免疫小鼠血清对HCVpp的中和活性与HVR1抗体水平呈正相关(r=0.967,P<0.01)。
表达截短E2蛋白的DNA疫苗可诱导抗HCV中和抗体,且中和抗体主要由抗HVR1抗体组成。