Development of effective mucosal vaccine against bacterial afferent infectious diseases

为了预防（疫苗）肠致病性大肠杆菌（ETEC）和副溶血性弧菌感染，重要的是刺激粘膜分泌性免疫球蛋白A（sIgA）的产生，以阻断肠上皮的定殖和中和毒素。重组霍乱毒素B亚单位（rCT B）被用作粘膜澄清剂。疫苗抗原的纯化利用His-tag融合载体（pQE-30 Xa）纯化ETEC菌毛定植因子抗原（CFA/I和CFA/III）的粘附素（CfaE和CfaJ）和副溶血性弧菌耐热直接溶血素（mTDH）的低溶血突变毒素.实验动物的免疫应答：BALB/c小鼠鼻内注射CfaE或CfaJ，可诱导小鼠产生CfaE和CfaJ特异性血清IgG和唾液、鼻腔、肺、阴道、用mTDH鼻内免疫的BALB/c小鼠，无论是否存在rCTB，均显示TDH特异性血清IgG抗体应答和粘膜免疫应答（sIgA）。尤其是在肺和小肠中。此外，免疫小鼠可以保护免受野生型TDH的致死性攻击。

For prevention (vaccine) of enterotoxigenic Escherichia coli (ETEC) and Vibrio parahaemolyticus infections, it is important to stimulate the production of mucosal secretory immunoglobulin A (sIgA) for blocking of colonization to intestinal epithelium and neutralizing of toxins. Recombinant cholera toxin B subunit (rCTB) was used as a mucosal adjuvant.1. Purification of vaccine antigenAdhesins (CfaE and CofJ) of pilus colonization factor antigens (CFA/I and CFA/III) of ETEC and less hemolytic mutant toxin of thermostable direct hemolysin (mTDH) of V. parahaemolyticus were purified by using His-tag fusion vector (pQE-30Xa).2. Immune response in experiment animalIntranasal administration of CfaE or CofJ to BALB/c mice with and without rCTB induced CfaE-and CofJ-specific serum IgG and mucosal immune response (sIgA) in the saliva, the nasal cavity, the lung, the vagina, the small intestine and the feces.BALB/c mice immunized intranasally with mTDH irrespective of the presence of rCTB showed TDH-specific serum IgG antibody response and mucosal immune response (sIgA), especially in the lung and the small intestine. Moreover, mice immunized could protect against lethal challenge with wild-type TDH.