CAJ | 학술논문

目的了解原核表达的登革病毒(dengue virus, DV)的E蛋白结构域Ⅲ直接抑制登革病毒感染及其抗体的中和作用.方法在大肠埃希菌中表达1~4型登革病毒E蛋白结构域Ⅲ(EⅢ).重组蛋白纯化后,进行阻断DV-2感染BHK~21细胞试验.用重组蛋白制备免疫血清,检测抗体中和作用.结果在大肠埃希菌中成功表达了1-4型登革病毒E蛋白结构域埃希菌,4型重组E蛋白结构域Ⅲ均能够阻断2型DV感染,4型重组蛋白的免疫血清均能中和2型DV,但中和抗体效价不同.结论原核表达的登革病毒结构域Ⅲ可以直接抑制病毒感染,所产生的抗体具有中和作用.直接抑制和中和抗体均对同型病毒作用较强.
목적 료해원핵표체적등혁병독(dengue virus, DV)적E단백결구역Ⅲ직접억제등혁병독감염급기항체적중화작용.방법 재대장애희균중표체1~4형등혁병독E단백결구역Ⅲ(EⅢ).중조단백순화후,진행조단DV-2감염BHK~21세포시험.용중조단백제비면역혈청,검측항체중화작용.결과 재대장애희균중성공표체료1-4형등혁병독E단백결구역애희균,4형중조E단백결구역Ⅲ균능구조단2형DV감염,4형중조단백적면역혈청균능중화2형DV,단중화항체효개불동.결론 원핵표체적등혁병독결구역Ⅲ가이직접억제병독감염,소산생적항체구유중화작용.직접억제화중화항체균대동형병독작용교강.
Objective To observe the ability of dengue virus recombinant envelope protein domain expressed in E. coil to inhibit virus infection and induce the neutralizing antibody. Methods E Ⅲ protein of Dengue virus serotypes 1-4 were expressed in E. coli BL21 (DE3) then purified. Recombinant proteins were tested to inhibit DV2 from infecting BHK-21 cell. Rabbits were immunized with recombinant proteins to produce anti-E Ⅲserum. Antibody titers were determined by neutralizing assay. Results The recombinant E Ⅲ proteins of Dengue virus serotypes 1-4 were expressed in E. coli. They effectively protected BHK cells in culture against DV2infection. All four type anti-E Ⅲ sera can neutralize DV2 but their efficacies are different. Conclusion E Ⅲproteins of dengue virus expressed in E. coli can directly inhibit DV2 infection. Neutralizing antibodies were induced by E Ⅲ proteins. Both E Ⅲ protein of dengue virus and the neutralizing antibodies they induced are more efficient in inhibiting homologous dengue serotypes infection than heterologous serotypes.