CAJ | 학술논문

本试验目的是研究牛IL-2重组质粒对口蹄疫病毒(Foot-and-mouth disease virus,FMDV)G-H环多肽免疫小鼠的佐剂效应.作者构建了含有信号肽序列的牛IL-2真核表达质粒pcDNA3.1-pBoIL-2,并体外鉴定其表达,同时原核表达并纯化了Asial型FMDV G-H环多肽.采用不同组合的pcDNA3.1-pBoIL-2、FMDV G-H环多肽分组免疫小鼠,并设阴、阳性对照,通过间接ELISA试验、微量血清中和试验、淋巴细胞增殖试验和实时定量PCR试验对各免疫组小鼠体液和细胞免疫水平进行综合评价.结果表明,与G-H+pcDNA3.1免疫组比较,G-H+pcD-NA3.1-pBoIL-2免疫组特异性抗体水平、中和抗体滴度、淋巴细胞增殖水平和细胞因子相对表达水平显著提高(P<0.05).结果提示,牛IL-2真核表达质粒可以显著提高G-H环多肽的免疫效果.
본시험목적시연구우IL-2중조질립대구제역병독(Foot-and-mouth disease virus,FMDV)G-H배다태면역소서적좌제효응.작자구건료함유신호태서렬적우IL-2진핵표체질립pcDNA3.1-pBoIL-2,병체외감정기표체,동시원핵표체병순화료Asial형FMDV G-H배다태.채용불동조합적pcDNA3.1-pBoIL-2、FMDV G-H배다태분조면역소서,병설음、양성대조,통과간접ELISA시험、미량혈청중화시험、림파세포증식시험화실시정량PCR시험대각면역조소서체액화세포면역수평진행종합평개.결과표명,여G-H+pcDNA3.1면역조비교,G-H+pcD-NA3.1-pBoIL-2면역조특이성항체수평、중화항체적도、림파세포증식수평화세포인자상대표체수평현저제고(P<0.05).결과제시,우IL-2진핵표체질립가이현저제고G-H배다태적면역효과.
To study the immune response induced by G-H loop peptide of Asial foot-and-mouth disease virus (FMDV) with BOIL-2 recombinant plasmid as adjuvant, the immune effects in im-munized mice were investigated. Recombinant plasmids pcDNA3.1-pBoIL-2 was constructed and the condition of the expression was identified in vitro. G-H peptide was expressed in Rosetta (DE3)pLysS and were successfully purified through GST affinity purification. Five groups of Kunming mice each with 8 were inoculated with different combinations including PBS, pcD-NA3.1-pBoIL-2 and/or G-H peptide, subsequently, immune effects have been evaluated through indirect ELISA assay, the neutralization test, lymphocytes proliferation assay and Quantitive Re-al-time PCR. The results showed that in comparing with the group immunized with G-H+pcD-NA3.1, the group communication with pcDNA3.1-pBoIL-2 induced significantly humoral and cellular immune responses. The results of this study indicate that the co-inoculation of G-H loop peptide with rBoIL-2 (as adjuvant) showed good immune effect.