表达 HIV-1建立感染相关病毒（T／F）膜蛋白的重组痘病毒与蛋白疫苗联合免疫豚鼠后体液免疫特征研究

摘要

目的：研究表达HIV-1建立感染相关病毒（ T／F）膜蛋白的重组痘苗病毒rTT-B、rTT-C和rTT-CON与gp140蛋白疫苗联合免疫豚鼠后抗体反应情况。方法采取Prime-boost免疫策略对豚鼠进行rTT初免1次，间隔4、8周后gp140蛋白疫苗加强免疫2次。免疫前和末次免疫后2、6、10周采豚鼠血，检测血清中HIV-1特异性结合抗体、中和抗体以及抗体的相对亲和力。结果（1） rTT-B、rTT-C和rTT-CON三种病毒和蛋白联合免疫均诱导了强烈的针对HIV-1 B′／C、B、AE亚型特异性结合抗体，抗体滴度范围为111430～1024000，其中，rTT-C和rTT-CON和蛋白联合免疫2周后诱导的B′／C和AE亚型特异性抗体滴度显著高于rTT-B（P＜0．05），但3种病毒蛋白联合免疫诱导的B亚型膜蛋白特异性抗体滴度差异无统计学意义。（2）3种病毒蛋白联合免疫都能诱导较强滴度针对SF162和ZM109的中和抗体，抗体滴度范围为83．76～649．30，3种病毒间差异无统计学意义。（3）3种病毒蛋白联合免疫都能诱导与免疫保护有关的HIV-1 V1V2-gp70特异性抗体，3种病毒间差异无统计学意义。（4）3种病毒蛋白联合免疫诱导的抗体对HIV-1 B′／C、B、AE亚型膜蛋白具有较强亲和力，3种病毒间差异无统计学意义。结论重组痘苗病毒rTT-B、rTT-C和rTT-CON与gp140联合免疫豚鼠后可以有效诱导出针对同源和异源病毒膜蛋白的结合及中和抗体。%Objective To analyze the antibody responses in guinea pigs vaccinated with recombi-nant vaccinia virus( rTT) strains expressing transmitted/founder ( T/F) HIV-1 membrane proteins in combi-nation with gp140 protein.Methods Guinea pigs were primed with rTT strains and boosted twice with gp140 protein in every four weeks.Serum samples were collected from guinea pigs before immunization and in 2, 6 and 10 weeks after the last immunization for the detection of HIV-1-specific binding antibodies, neu-tralizingantibodiesandtherelativeavidityofantibodies.Results (1)Thebindingantibodiesspecificto HIV-1 B′/C, B, AE subtypes were efficiently induced by the immunization of rTT-B, rTT-C and rTT-CON vaccinia strains in combination with gp140 protein.The antibody titers ranged from 111 430 to 1 024 000. More antibodies against HIV-1 B′/C and AE subtypes were induced in guinea pigs by the immunization of rTT-C and rTT-CON strains in combination with gp140 protein than those by using rTT-B strain prime-protein boost strategy (P<0.05).No significant differences with the titers of HIV-1 B subtype specific antibody were observed among the guinea pigs immunized with the three strategies.( 2 ) High titers of SF162 and ZM109 neutralizing antibodies were induced in guinea pigs immunized with rTT-B, rTT-C and rTT-CON vac-cinia strains in combination with gp140 protein, ranging from 83.76 to 649.30.No significant differences were found among the three groups.(3) The HIV-1 V1V2-gp70 specific antibodies associated with protec-tive immunity were induced by immunization of the three virus prime-protein boost strategies.No significant differences were observed among them.(4) Antibodies induced in guinea pigs by immunization of the three strategies showed strong affinity to membrane proteins of HIV-1 B′/C, B, AE subtype strains.No significant differences were found among the three immunization strategies.Conclusion A strong humoral immune re-sponse was induced in guinea pigs primed with recombinant vaccinia virus strains expressing T/F virus HIV-1 membrane proteins and boosted with gp140 protein.