CAJ | 학술논문

目的构建登革病毒1～4型重组亚单位疫苗,分析重组疫苗的免疫原性.方法利用连接肽将登革病毒1型与2型,3型与4型的外膜蛋白DⅢ基因片段连接在一起,克隆人原核表达载体pET-30a,在大肠埃希菌中表达DEN1/2型、DEN3/4型融合重组蛋白.重组蛋白经高效液相色谱柱分离纯化后,混和免疫小鼠,采用中和实验法测定血清中和抗体效价,同时进行乳鼠体内中和实验,观察中和抗体对乳鼠的免疫保护作用.结果重组蛋白能诱导小鼠产生针对登革病毒1～4型的中和抗体,平均中和效价分别为1:34.9、1:45.3、1:24.7和1:38.4.中和抗体能保护新生乳鼠免受致死剂量DEN1～4的攻击,保护率分别为100%,100%,83%,83%.结论构建的重组亚单位疫苗具有良好的免疫原性,能诱导小鼠产生中和抗体,为构建单一四价疫苗提供了实验室依据.
목적 구건등혁병독1～4형중조아단위역묘,분석중조역묘적면역원성.방법 이용련접태장등혁병독1형여2형,3형여4형적외막단백DⅢ기인편단련접재일기,극륭인원핵표체재체pET-30a,재대장애희균중표체DEN1/2형、DEN3/4형융합중조단백.중조단백경고효액상색보주분리순화후,혼화면역소서,채용중화실험법측정혈청중화항체효개,동시진행유서체내중화실험,관찰중화항체대유서적면역보호작용.결과 중조단백능유도소서산생침대등혁병독1～4형적중화항체,평균중화효개분별위1:34.9、1:45.3、1:24.7화1:38.4.중화항체능보호신생유서면수치사제량DEN1～4적공격,보호솔분별위100%,100%,83%,83%.결론 구건적중조아단위역묘구유량호적면역원성,능유도소서산생중화항체,위구건단일사개역묘제공료실험실의거.
Objective To construct sub-unit vaccines of dengue virus type 1 to 4 and to analyze its immunogenicity. Methods Envelope domain Ⅲ s of dengue serotypes 1 and 2, as well as 3 and 4, were spliced by a linker (Gly-Gly-Ser-Gly-Ser) 3 and cloned into vector pET-30a, then transformed into E. coli to express recombinant fusion proteins. The recombinant proteins were purified by high-performance liquid chromatography and mixed to immunize BALB/c mice. The neutralizing antibodies were tested by neutralizing assay, as well as in newborn mice challenged intracranially with dengue virus type 1 to 4.combination with sera from mice immunized with recombinant proteins were protected, whereas 83% protection was obtained when challenged with DEN-3 or 4. Conclusion The recombinant proteins possess excellent immunogenicity to induce neutralizing antibodies and would be valuable for development of a tetravalent sub-unit vaccine.