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[目的]用人巨细胞病毒(HCMV)IE1核酸疫苗pcDNA3.1(-)-IE1免疫BALB/c小鼠,初步研究其产生的体液免疫和细胞免疫应答水平。[方法]将pcDNA3.1(-)-IE1通过肌肉注射免疫BALB/c小鼠,通过PCR测定和免疫组化检测其在肌细胞中的表达,细胞因子测定、淋巴细胞转化实验检测免疫效果。[结果]PCR检测到与IE1大小一致的片段,免疫组化结果显示IE1基因在小鼠肌细胞中表达IE1目的蛋白。小鼠脾淋巴细胞经PHA刺激后,实验组IL-4、IL-2、IFN-γ含量以及淋巴细胞转化率显著增高,与对照组比较差异有统计学意义(P﹤0.05)。[结论]pcDNA3.1(-)-IE1核酸疫苗能在BALB/c小鼠肌细胞中稳定存在并能表达HCMV IE1蛋白,pcDNA3.1(-)-IE1核酸疫苗可诱导BALB/c小鼠脾细胞分泌IL-4、IL-2、IFN-r并刺激BALB/c小鼠脾细胞增殖。
[Objective] To study the humoral and cellular immune response of BALB / c mice immunized with human cytomegalovirus (HCMV) IE1 DNA vaccine pcDNA3.1 (-) - IE1. [Method] The BALB / c mice were immunized with pcDNA3.1 (-) - IE1 by intramuscular injection. The expression of pcDNA3.1 (-) - IE1 was detected by PCR and immunohistochemistry in muscle cells. Cytokines and lymphocyte transformation were used to test the immune effect. [Result] The fragment with the same size as IE1 was detected by PCR. The results of immunohistochemistry showed that IE1 gene expressed IE1 protein in mouse muscle cells. After stimulated by PHA, the levels of IL-4, IL-2 and IFN-γ in experimental splenic lymphocytes were significantly increased as compared with the control group (P <0.05). [Conclusion] The pcDNA3.1 (-) - IE1 DNA vaccine can stably exist in myocytes of BALB / c mice and express the HCMV IE1 protein. The pcDNA3.1 (-) - IE1 DNA vaccine can induce the splenic Cells secrete IL-4, IL-2, IFN-r and stimulate splenocyte proliferation in BALB / c mice.