空肠弯曲菌PEB1重组蛋白的免疫原性研究

目的探讨空肠弯曲菌PEB1重组蛋白的免疫原性及对机体的保护作用。方法以不同剂量的含弗氏佐剂的纯化PEB1重组蛋白为疫苗,分别经皮下注射和肌肉注射的方法免疫Balb/C小鼠,ELISA法测定血清中特异性抗体的含量,MTT法测定细胞增殖效应。CJ菌液攻击实验,观察攻击后小鼠的精神状态、病死情况,并做血液、肠道分泌液细菌培养。结果用PEB1重组蛋白免疫Balb/C小鼠,可诱导出高滴度的特异性抗体,能有效地刺激淋巴细胞增殖,加入刺激原的各实验组SI值均较对照组显著增高。结论PEB1重组蛋白免疫接种后可诱导出高滴度特异性抗体和致敏淋巴细胞,能有效保护小鼠免受大剂量空肠弯曲菌的攻击,是一种比较理想的基因工程亚单位疫苗候选抗原。细菌攻击实验结果表明,PEB1基因工程亚单位疫苗能使实验动物产生有效的免疫保护力,明显降低动物的患病率和病死率,并能有效地降低血液和肠道分泌液的细菌含量。

Immunogenicity of recombinant PEB1 protein of campylobacter jejuni

Objective To explore the immunogenicity and protection of campylobacter jejuni （CJ） recombinant PEB1 protein. Methods Balb/C mice were immunized by different doses of the purified PEB1 protein with Freund＇ s adjuvant using subcutaneous injection or intramuscular injection ELISA was used to detect the specific antibody titer and MTT method was used for measuring the stimulation index （SI） of the spleen lymphocyte transformation. The animals were observed for the incidence and death of sickness after attacking with（CJ）. CJ bacteria cultures were also performed from intes- tinal excreting fluid and blood sample. Results The specific IgG antibody was found in serum of Balb/C mice immunized by recombinant PEB1 protein. The proliferating responses of immunized mice spleen lymphocyte were induced when stimula- ted by PHA or PEB1 protein, and the SI value of immunized groups were significantly higher than control group. Effective immunological protection with lower sickness incidence and mortality was seen in the mice suffered from massive （C J） infection. The number of （CJ） in intestine and blood was reduced. Conclusions Effective humoral immunity and cell media- ted immunity was induced after immunization by recombinant PEB1 protein. The Balb/C mice immunized with recombinant PEB1 protein get survival from high dose CJ infection. It suggests the recombinant PEB1 protein is a valuable candidate for campylobacter jejuni genetic engineering subunit vaccine.