摘要
目的初步研究日本血吸虫大陆株副肌球蛋白核酸疫苗在C57BL/6小鼠诱生的免疫应答特征。方法利用TRIzol方法抽提日本血吸虫大陆株成虫总RNA,并通过逆转录聚合酶链反应;扩增编码日本血吸虫大陆株副肌球蛋白(SjC97)的全基因cDNA,将其克隆入质粒表达载体(pcDNA/Amp)构建成日本血吸虫大陆株副肌球蛋白全基因核酸疫苗(pCMV-SjC97);将其通过后腿胫前肌免疫C57BL/6小鼠,共免疫3次,每次间隔3周;利用酶联免疫吸附试验(ELISA)分析pCMV-SjC97免疫所诱生的特异性抗体IgG及IgG亚类,以及特异性抗体IgM及IgE。并初步分析了pCMV-SjC97免疫C57BL/6小鼠脾淋巴细胞抗原特异性体外增殖能力。结果pCMV-SjC97免疫能够诱生特异性抗体IgG及IgM;pCMV-SjC97诱生的抗体滴度与注射的DNA间存在一定的量-效关系;IgG亚类分析显示,pCMV-SjC97免疫主要诱生IgG2a和IgG2b亚类,未测到IgG1及IgG3亚类。PCMV-SjC97免疫能够诱导小鼠Th细胞激活。结论pCMV-SjC97核酸疫苗能够在C57BL/6小鼠诱生较强的体液免疫应答;pCMV-SjC97免疫主要诱生Th1型细胞免疫应答。
Objective To study the characteristics of immune response in C57BL/6 mice by nucleic acid encoding full-length paramyosin of Chinese S. japonicum. Methods Total RNA was isolatal from adult Chinese S. japonicum using TRIzol reagent. The full-length cDNA encoding paramyosin of Chinese S. japonicum was amplified by RT-PCR and cloned into expression vector pcDNA/Amp (pCMV-SjC97), which was intramuscularly injected into C57 BL/6 mice. The mice were vaccinated with DNA the three-week interval for a total of three inoculations. The specific anti-SjC97 IgM, IgG and IgE antibodies and IgG subclasses were determined by ELISA test. Results The pCMV-SjC97 vaccine induced specific anti-SjC97 IgM and IgG antibodies. The antibody resporise correlated well with the administered amount of pCMV-SjC97. The IgG subclasses, predominantly IgG2a and IgG2b, were observed in the mice vaccinated with pCMV-SjC97, but no IgG1 and IgG3 responses were seen.Conclusion The strong immune response, especially Th1 type, can be induced in the mice with pCMV-SjC97 vaccine.
出处
《中华医学杂志》
CAS
CSCD
北大核心
1999年第11期848-851,共4页
National Medical Journal of China
基金
中国博士后基金!986
国家863高技术计划资助!102070401
TMRC/NIH部分资助!50 AI39461
关键词
日本血吸虫
原肌球蛋白
疫苗
核酸疫苗
Schistosoma japonicum
Tropomyosin
Vaccines
Immune active