摘要
目的对小鼠进行两种基因免疫方法的比较:分别以真核质粒pcDN3.1(+)为载体和以伤寒减毒活菌苗为载体,携带丙肝病毒高变区1(HVR1)相关模拟表位的DNA序列,诱导细胞免疫应答,以寻找较好的疫苗免疫途径。方法根据HCV HVR1模拟表位的肽序列合成DNA序列,并将其连接到pcDN3.1(+)(pcDN3.1-SP)真核质粒上,然后用重组质粒转化伤寒减毒活菌苗Ty21a(Ty21a-SP)。Ty21a-SP和pcDN3.1-SP分别经口服和肌注免疫小鼠后,杀鼠、分离脾细胞,脾细胞经混合肽刺激后,用流式细胞仪检测CD8+IFN-γ+细胞;用非放射性MTS法检测细胞增殖反应;以非放射性LDH法检测CTL反应。结果对小鼠用pcDN3.1-SP和Ty21a-SP免疫后,取脾淋巴细胞经混合肽刺激后,明显增殖,CD8+IFNγ-+淋巴细胞比例增高,并诱导较强的CTL反应,但pcDN3.1-SP免疫后的上述反应较Ty21a-SP免疫弱。结论使用伤寒减毒活菌苗作为载体进行基因免疫有利于产生细胞免疫反应。
Objective To compare the cellular immune responses induced by two different vectors, eukaryotic plasmid pcDN3. 1（＋） and attenuated typhia live vaccine Ty21a, -associated mimic epitopes of HCV hypervariable region 1, so to find a better way for vaccine immunization. Methods The DNA sequence based on the peptides sequence of HCV hypervariable region I related consensus polymimotopas was synthesize and then inserted into plasmid pcDN3. 1 （＋） to construct recombinant plasmid pcDN3. 1 （＋）-SP, which then was transfected into live attenuated typhia vaccine Ty21a to construct Ty21a-SP. The mice were immunized orally with Ty21a-SP and intramuscularly with peDN3. 1（＋）-SP, respectively, and then sacrificed by exsanguination. The splenocytes were separated and restimulated with pooled synthesized peptides, and then collected. Flow cytometry was employed to identify CD6 ＋ IFN-γ＋ T cells, and non-radioactive MTS method was adopted to test T cell proliferation, and non-radioactive LDH method was used to test cytotoxic T cytolytic reaction （CTL）. Results Compared with control groups, the proliferation of splenocytes was apparently enhanced, the proportion of CD8＋IFN-γ＋ cells obviously increased, and CTL responses also significantly increased after the spenocytes of mice immunized by pcDN3. 1-SP and Ty21a-SP were restimulated with synthesized peptides. And the responses mentioned above in the mice immunized by Ty21a-SP were stronger than those mice immunized by peDN3. 1 SP. Conclusion Using attenuated typhia live vaccine Ty21a as DNA vector is an effective way to induce cellular immune responses
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2008年第10期1202-1204,共3页
Medical Journal of Chinese People's Liberation Army
关键词
模拟表位
伤寒疫苗
细胞免疫反应
基因免疫
mimotopes
attenuated typhia live vaccine
cellular immune responses
gene immunization
