论文部分内容阅读
目的:研究空肠弯曲菌pcDNA3.1(-)-peb1A壳聚糖佐剂疫苗的免疫原性和保护性。方法:健康雄性昆明小鼠分为实验组和对照组。实验组设pcDNA3.1(-)-peb1A组3个剂量和壳聚糖-pcDNA3.1(-)-peb1A组3个剂量;对照组设空载体pcDNA3.1(-)(100μg/100μl)组和生理盐水(NS)组,各组均采用小鼠股四头肌注射法。在第0、10、20天免疫,于每次免疫后第10天采集各组小鼠血清,以间接ELISA法检测血清中特异性IgM、IgG的含量。分离各组小鼠脾淋巴细胞包被细胞培养板,以细胞ELISA法检测小鼠脾淋巴细胞CD20、CD21表达水平。设空肠弯曲菌液灌胃攻击免疫后小鼠,检测肠道分泌液细菌培养数量。结果:裸DNA组和壳聚糖-DNA组各组特异性IgG水平均高于两对照组(P<0.05)。裸DNA组和壳聚糖-DNA组小鼠脾淋巴细胞CD20、CD21表达水平均高于两对照组(P<0.05),壳聚糖-DNA组高于裸DNA组(P<0.05)。肠道分泌液细菌培养结果细菌指数裸DNA组的低于两对照组;佐剂DNA组低于裸DNA组。结论:重组质粒pcDNA3.1(-)-peb1A经肌肉注射免疫小鼠,具有较好的免疫原性和保护作用,壳聚糖能增强pcDNA3.1(-)-peblA的免疫原性,有望成为空肠弯曲菌基因疫苗的候选佐剂。
Objective: To study the immunogenicity and protective effect of jejunum jejuni pcDNA3.1 (-) - peb1A chitosan adjuvant vaccine. Methods: Healthy male Kunming mice were divided into experimental group and control group. Three doses of pcDNA3.1 (-) - peb1A and three doses of chitosan-pcDNA3.1 (-) - peb1A were set in the experimental group and the blank vector pcDNA3.1 (-) (100μg / 100μl) And normal saline (NS) group, all mice were injected quadriceps femoris. The mice were immunized on days 0, 10 and 20, and the serum of each group was collected on the 10th day after each immunization. The contents of specific IgM and IgG in serum were detected by indirect ELISA. The splenic lymphocytes of each group were separated and coated on the cell culture plate. The expression of CD20 and CD21 in splenic lymphocytes of mice was detected by ELISA. Set the Campylobacter jejuni gavage challenge immunized mice to detect intestinal secretion bacterial culture quantity. Results: The specific IgG levels in each group of naked DNA group and chitosan-DNA group were higher than those of the two control groups (P <0.05). The expression of CD20 and CD21 in spleen lymphocytes of nude DNA group and chitosan-DNA group were higher than that of the two control groups (P <0.05), chitosan-DNA group was higher than that of naked DNA group (P <0.05). Bacterial culture results of intestinal excretion Bacterial index was lower in the nude DNA group than in the two control groups; adjuvant DNA group was lower than in the nude DNA group. CONCLUSION: The recombinant plasmid pcDNA3.1 (-) - peb1A is immunized by intramuscular injection and has good immunogenicity and protective effect. Chitosan can enhance the immunogenicity of pcDNA3.1 (-) - peblA and is expected to become Candidate adjuvant for Campylobacter jejuni genetic vaccine.