CAJ | 학술논문

目的研究细胞因子佐剂白介素23(IL-23)和佐剂CpG2216对呼吸道合胞病毒(RSV)重组疫苗G1F/M2免疫原性的影响.方法将编码IL-23的质粒(简称IL-23)单独作为佐剂以及将IL-23和CpG2216联合作为佐剂与G1F/M2共免疫BABL/c小鼠三次,在每次免疫后10 d取血,分离血清,用间接ELISA法测定三次免疫后的血清IgG抗体效价,观察佐剂对体液免疫的影响.第3次免疫后2周处死小鼠,取脾细胞,用LDH法检测CTL活性、ELISPOT法检测分泌IL-4和γ-干扰素(IFN-γ)的淋巴细胞水平、流式细胞仪检测CD4+、CD8+的效应记忆和中央记忆T细胞,观察佐剂对细胞免疫的影响.结果 ELISA结果表明:除PBS组外,各佐剂组均诱导了高滴度的IgG抗体,其中IL-23+G1F/M2组的IgG水平显著低于PBS+G1F/M2组和IL-23+CpG2216+G1F/M2组,另外,PBS+G1F/M2组与IL-23+CpG2216相比差异无统计学意义;CTL杀伤实验显示:IL-23+CpG2216+G1F/M2组的CTL杀伤活性显著高于其他组;ELISPOT结果显示IL-23+CpG2216+G1F/M2组分泌IFN-γ淋巴细胞数量显著多于其他组,且各组分泌IFN-γ的淋巴细胞数显著多于分泌IL-4的淋巴细胞;流式细胞仪检测结果表明,IL-23和IL-23+CpG2216对G1F/M2诱导产生记忆细胞有明显增强作用.结论佐剂IL-23和佐剂CpG2216联合使用能够增强RSV重组疫苗的免疫应答.
목적 연구세포인자좌제백개소23(IL-23)화좌제CpG2216대호흡도합포병독(RSV)중조역묘G1F/M2면역원성적영향.방법 장편마IL-23적질립(간칭IL-23)단독작위좌제이급장IL-23화CpG2216연합작위좌제여G1F/M2공면역BABL/c소서삼차,재매차면역후10 d취혈,분리혈청,용간접ELISA법측정삼차면역후적혈청IgG항체효개,관찰좌제대체액면역적영향.제3차면역후2주처사소서,취비세포,용LDH법검측CTL활성、ELISPOT법검측분비IL-4화γ-간우소(IFN-γ)적림파세포수평、류식세포의검측CD4+、CD8+적효응기억화중앙기억T세포,관찰좌제대세포면역적영향.결과 ELISA결과표명:제PBS조외,각좌제조균유도료고적도적IgG항체,기중IL-23+G1F/M2조적IgG수평현저저우PBS+G1F/M2조화IL-23+CpG2216+G1F/M2조,령외,PBS+G1F/M2조여IL-23+CpG2216상비차이무통계학의의;CTL살상실험현시:IL-23+CpG2216+G1F/M2조적CTL살상활성현저고우기타조;ELISPOT결과현시IL-23+CpG2216+G1F/M2조분비IFN-γ림파세포수량현저다우기타조,차각조분비IFN-γ적림파세포수현저다우분비IL-4적림파세포;류식세포의검측결과표명,IL-23화IL-23+CpG2216대G1F/M2유도산생기억세포유명현증강작용.결론 좌제IL-23화좌제CpG2216연합사용능구증강RSV중조역묘적면역응답.
Objective To investigate the role of IL-23 and CpG2216 adjuvant in the cellular immune response induced by recombinant protein vaccine of respiratory syncytial virus (RSV). Methods BABL/c mice were immunized with RSV recombinant vaccines G1F/M2 protein with pcDNA3. l-IL-23 eukaryotic expression plasmid(IL-23) combined or alone with CpG2216 as immunologic adjuvant. An interval of 10 days was invariably employed for second, and third immunizations. Mice were phlebotomized 10 days after every immunization and the serum was separated. Specific antibody IgG titers in sera from vaccinated mice were assessed by indirect ELISA. Spleens from the mice immunized were removed two weeks after the third immunization. Specific lysis activity was assessed using a LDH Cytotoxicity Assay Kit. IFN-γ- and IL-4-secreting cells were quantified using a ELISPOT kit. Percentages of CD4+/CD8+ effector cells or memory cells in the spleen cells of mice were analyzed with flow cytometer. Results The mice of all groups induced specific IgG antibodies compared with PBS group after the third immunization ( P <0. 05),except the level of IgG in IL-23 + G1F/M2 group is the lowest. The specific lysis activity induced by IL-23+ CpG2216+ G1F/M2 was significantly higher than other groups. IL-23 + CPG2216+G1F/M2 group, induced significantly more IFN-γ secreting cells than other groups ( P <0. 05). In addition, the level of IFN-γ-secreting cells was remarkably higher than that of IL-4-secreting cells in each group. Both IL-23 and IL-23 + CpG2216 have significant advantages to enhanced immune memory response. Conclusions IL-23 + CpG2216 , as an adjuvant, can enhanced significantly the cellular immune responses of RSV recombinant vaccine G1F/M2.