CAJ | 학술논문

为了筛选大肠杆菌疫苗的有效抗原，本研究表达了重组蛋白Stx2B和重组蛋白LTB-Stx2B，并分别用重组蛋白Stx2B、表达了重组蛋白LTB-Stx2B的灭活菌液和灭活菌裂解液免疫昆明鼠，比较了不同抗原诱导小鼠的体液免疫应答能力；体内体外实验对免疫血清的毒素中和作用进行了检测。结果显示，重组蛋白LTB-Stx2B能诱导小鼠更高水平的体液免疫应答，其血清抗体可以通过乳汁传递给新生仔鼠；小鼠二免后的血清不仅能中和LT和Stx2对小鼠的毒性作用，而且能中和Stx2对Vero细胞的毒性，其中和效价为1：52，高于其他免疫原组血清。该研究为大肠杆菌疫苗的研究奠定了基础。
위료사선대장간균역묘적유효항원，본연구표체료중조단백Stx2B화중조단백LTB-Stx2B，병분별용중조단백Stx2B、표체료중조단백LTB-Stx2B적멸활균액화멸활균렬해액면역곤명서，비교료불동항원유도소서적체액면역응답능력；체내체외실험대면역혈청적독소중화작용진행료검측。결과현시，중조단백LTB-Stx2B능유도소서경고수평적체액면역응답，기혈청항체가이통과유즙전체급신생자서；소서이면후적혈청불부능중화LT화Stx2대소서적독성작용，이차능중화Stx2대Vero세포적독성，기중화효개위1：52，고우기타면역원조혈청。해연구위대장간균역묘적연구전정료기출。
The objective of the present study was to investigate into immunogenic antigens for use in E.coli vaccines. The recombinant Rossetta/pCold I-LTB-Stx2B was expressed in E. coli with induction of IPTG. The cells were treated with sonication and lysates containing recombinant proteins Stx2B and LTB-Stx2B were inactivated. The lysates were used to immunize Kunming mice. The humoral immune responses of vaccinated mice were examined. The neutralizing effects of serum antibodies against LT and Stx2 were tested in vivo and in vitro. The results showed that the recombinant protein LTB-Stx2B induced a higher level of humoral immune response in vaccinated mice. Besides, specific antibodies were transferred to newborn mice through the colostrums. The serum samples collected after the second immunization protected mice from the challenge with LT and Stx2 and neutralized the toxicity of Stx2 in mice and on Vero cells. The neutralizing titer was up to 1:52 tested on Vero cell. This study provided preliminary information on development of efficient E. coli vaccines.