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目的应用原核表达系统制备IL-1β重组蛋白,并作为流感喷鼻疫苗的佐剂,研究其作为喷鼻疫苗佐剂的有效性。方法从Genbank获得IL-1β的基因cDNA序列,在大肠杆菌中表达。分离纯化表达产物,与甲型H1N1流感喷鼻疫苗均匀混合,免疫Balb/c小鼠,通过对体液免疫、黏膜免疫和细胞免疫水平的检测,证明rIL-1β蛋白佐剂的有效性。结果 rIL-1β蛋白在大肠杆菌中获得了高效表达,表达量约是30%,纯度是94.5%。制备的rIL-1β蛋白具有无毒、稳定和良好的生物活性。通过对小鼠的免疫实验证明,rIL-1β作为喷鼻疫苗佐剂对体液、黏膜和细胞免疫应答均有良好的增强效果。结论 rIL-1β蛋白具有黏膜佐剂的作用,为深入研究佐剂效应提供了实验数据。
OBJECTIVE: To prepare IL-1β recombinant protein by using prokaryotic expression system and to evaluate its effectiveness as an adjuvant for nasal spray vaccine. Methods The cDNA sequence of IL-1β was obtained from Genbank and expressed in E.coli. The expressed product was isolated and purified and mixed with the H1N1 influenza nasal spray vaccine to immunize Balb / c mice. The humoral immunity, mucosal immunity and cellular immunity levels were tested to prove the effectiveness of rIL-1β adjuvant. Results The rIL-1β protein was highly expressed in E. coli. The expression level was about 30% and the purity was 94.5%. The prepared rIL-1β protein has non-toxic, stable and good biological activity. Immunization experiments on mice demonstrated that rIL-1β has a good enhancing effect on humoral, mucosal and cellular immune responses as a nasal vaccine adjuvant. Conclusion The rIL-1β protein has mucosal adjuvant effect and provides experimental data for further study of adjuvant effect.