Hu Hui, Lu Xinya, Tao Ling, Bai Bingke, Zhang Zhenfeng, Chen Yao, Zheng Fangliang, Chen Jianjun, Chen Ze, Wang Hanzhong
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, People's Republic of China.
Clin Vaccine Immunol. 2007 Jul;14(7):894-901. doi: 10.1128/CVI.00019-07. Epub 2007 May 9.
DNA vaccines induce humoral and cellular immune responses in animal models and humans. To analyze the immunogenicity of the severe acute respiratory syndrome (SARS) coronavirus (CoV), SARS-CoV, spike DNA vaccine and the immunoregulatory activity of interleukin-2 (IL-2), DNA vaccine plasmids pcDNA-S and pcDNA-IL-2 were constructed and inoculated into BALB/c mice with or without pcDNA-IL-2 by using three different immunization routes (the intramuscular route, electroporation, or the oral route with live attenuated Salmonella enterica serovar Typhimurium). The cellular and humoral immune responses were assessed by enzyme-linked immunosorbent assays, lymphocyte proliferation assays, enzyme-linked immunospot assays, and fluorescence-activated cell sorter analyses. The results showed that specific humoral and cellular immunities could be induced in mice by inoculating them with SARS-CoV spike DNA vaccine alone or by coinoculation with IL-2-expressing plasmids. In addition, the immune response levels in the coinoculation groups were significantly higher than those in groups receiving the spike DNA vaccine alone. The comparison between the three vaccination routes indicated that oral vaccination evoked a vigorous T-cell response and a weak response predominantly with subclass immunoglobulin G2a (IgG2a) antibody. However, intramuscular immunization evoked a vigorous antibody response and a weak T-cell response, and vaccination by electroporation evoked a vigorous response with a predominant subclass IgG1 antibody response and a moderate T-cell response. Our findings show that the spike DNA vaccine has good immunogenicity and can induce specific humoral and cellular immunities in BALB/c mice, while IL-2 plays an immunoadjuvant role and enhances the humoral and cellular immune responses. Different vaccination routes also evoke distinct immune responses. This study provides basic information for the design of DNA vaccines against SARS-CoV.
DNA疫苗可在动物模型和人类中诱导体液免疫和细胞免疫反应。为分析严重急性呼吸综合征(SARS)冠状病毒(CoV)、SARS-CoV刺突DNA疫苗的免疫原性以及白细胞介素-2(IL-2)的免疫调节活性,构建了DNA疫苗质粒pcDNA-S和pcDNA-IL-2,并通过三种不同免疫途径(肌肉注射途径、电穿孔或用减毒活鼠伤寒沙门氏菌经口服途径)将其接种到BALB/c小鼠体内,接种时有的小鼠同时接种pcDNA-IL-2,有的不接种。通过酶联免疫吸附测定、淋巴细胞增殖测定、酶联免疫斑点测定和荧光激活细胞分选分析来评估细胞免疫和体液免疫反应。结果表明,单独接种SARS-CoV刺突DNA疫苗或与表达IL-2的质粒共接种均可在小鼠中诱导特异性体液免疫和细胞免疫。此外,共接种组的免疫反应水平显著高于单独接种刺突DNA疫苗的组。三种接种途径的比较表明,口服接种引发强烈的T细胞反应,主要产生较弱的免疫球蛋白G2a(IgG2a)亚类抗体反应。然而,肌肉注射引发强烈抗体反应和较弱的T细胞反应,电穿孔接种引发强烈反应,主要产生IgG1亚类抗体反应和中等强度的T细胞反应。我们的研究结果表明,刺突DNA疫苗具有良好的免疫原性,可在BALB/c小鼠中诱导特异性体液免疫和细胞免疫,而IL-2起免疫佐剂作用,可增强体液免疫和细胞免疫反应。不同的接种途径也会引发不同的免疫反应。本研究为抗SARS-CoV DNA疫苗的设计提供了基础信息。