人巨细胞病毒IE1核酸疫苗的免疫效果研究  

IMMUNOCOMPETENCE STUDY OF HUMAN CYTOMEGALOVIRUS IE1 DNA VACCIN IN MICE

作　　者：刘安元[1] 曹清香[1] 唐双阳[1] 陈琳[1] 万艳平[1] 

机构地区：[1]湖南省南华大学病原生物学研究所,衡阳421001

出　　处：《现代预防医学》2011年第9期1709-1712,共4页Modern Preventive Medicine

基　　金：湖南省自然科学基金项目(06JJ4034);湖南省卫生厅课题项目(B2004-077)

摘　　要：[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 observe the humoral or cellular immune responses,the HCMV IE1 DNA vaccine is injected by intramuscular cells in BALB/c mice,and it is expected to provide experimental basis for further studying DNA vaccine against HCMV infection.[Methods]BABL/c mice were immunized with pcDNA3.1（-）-IE1 or pcDNA3.1（-） or PBS buffer by intramuscular cells at 2-week interval for four times.Two weeks after the last time of immunization,HCMV IE1 gene or protein in intramuscular cells was detected by PCR or observed with immunohistochemistry.Two weeks after the last time of immunization,ELISA was used to detect the quantities of IL-4,IL-2 or IFN-γ in the supernatant of murine spleen lymphocyte culture after stimulated by PHA.The proliferation response of spleen cells was detected using MTT assay,and the stimulation indexes（SI） were counted.All of data were analyzed by SPSS13.0.[Results]In the intramuscular cells of immunized mice,the fragments were consistent to HCMV IE1 gene detected by PCR.The result of immunohistochemistry showed that HCMV IE1 gene could express in intramuscular cells of mice.CKs in mice inoculated with pcDNA3.1（-）-IE1 were much more than those with pcDNA3.1（-） or with PBS,it was significant difference between the experiment group and control group（IL-4：P﹤0.05,IL-2 and IFN-γ：P﹤0.01）;The stimulation index（SI） of spleen cells of DNA vaccine group was significantly higher than those of pcDNA3.1（-） group or PBS group（P﹤0.01） .[Conclusion]HCMV IE1 gene could exist constantly and express in intramuscular cells of BALB/c mice.The level of IL-4,IL-2 or IFN-r and stimulation indexs of spleen cells could be increased in BALB/c mice immunized with pcDNA3.1（-）-IE1.

关 键 词：人巨细胞病毒(HCMV) IE1 疫苗 免疫应答 

分 类 号：R186[医药卫生—流行病学]