摘要
为鉴定禽流感病毒核蛋白(NP)中的T细胞表位,本研究根据蛋白结构预测技术预测了4个潜在细胞毒性T淋巴细胞(CTL)表位(NP120-128、NP163-172、NP85-92、NP67-74),并合成相应的候选短肽。构建了含有NP基因的重组质粒p CAGGS-NP,将其转染293T细胞。间接免疫荧光和western blot结果表明NP蛋白能够在293T细胞中获得表达。重组质粒免疫SPF鸡后能够诱导鸡体产生特异的NP抗体。在合成的4条短肽中,NP67-74体外刺激免疫过p CAGGS-NP的鸡脾淋巴细胞可以显著提高脾淋巴细胞IFN-γ的分泌量,提示短肽NP67-74能够在体外诱导活化鸡淋巴细胞产生CTL,可能是AIV NP的CTL表位。该研究结果对流感病毒免疫机制和通用疫苗研究具有借鉴意义。
In order to identify the T cell epitope in avian influenza virus （A1V） nucleoprotein （NP）, four T cell epitope candidate peptides （NP104-112 , NP163-172, NP85-92 and NP67-74） of the AIV NP were predicted with the protein structure prediction technology and synthesized. To verify the immunogenicity of these peptides, the recombinant plasmid pCAGGS-NP was constructed and the expression of NP was confirmed by indirect immunofluorescence and western blot in transfected 293T cell. The antibodies against NP were also detected by ELISA in pCAGGS-NP inoculated SPF chickens. The splenic cells were collected from inoculated chickens and stimulated with the peptides of NP120-128, NP163-172, NP85-92, NP67-74 or negative control, respectively. The results of real-time PCR show that spleen lymphocytes stimulated with the peptide NP67.74 significantly increased the IFN-7 secretion, which indicated that the peptide NP67-74 might be AIV NP CTL epitope. The results possessed significance to mechanism of protective immunity against influenza virus and the research towards universal flu vaccine.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2015年第11期861-865,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
现代农业产业技术体系(CARS-42-Z12)
公益性行业(农业)科研专项(201303033)
十二五国家科技支撑计划(2015BAD12B03)
