T细胞表位多肽疫苗对禽流感病毒的免疫保护评价
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摘要
为了探究NP蛋白T细胞表位多肽NP_(67-74)-KLH对流感通用疫苗4M2e-MAP免疫效果的影响,本研究将多肽NP_(67-74)-KLH和4M2e-MAP添加完全弗氏佐剂和不完全弗氏佐剂后免疫BALB/c小鼠,通过攻毒保护试验来评价免疫效果。攻毒以后小鼠体重变化曲线说明,联合免疫多肽疫苗NP_(67-74)-KLH和4M2e-MAP的试验组小鼠体重变化趋势比单一免疫4M2e-MAP合成肽疫苗更加趋于平稳,并且联合免疫试验组小鼠体重恢复较快。荧光定量和肺部组织病理切片结果均表明,复合多肽(NP_(67-74)-KLH和4M2eMAP)能在一定程度上干扰流感病毒在脏器肺中的复制并能够降低病毒对肺部的损伤程度,在抵抗病毒过程中可以产生有效保护。该研究为多肽疫苗联合应用以抵抗流感研发提供了较好思路。
To explore the effect of T cell epitope peptide NP_(67-74)- KLH of NP protein on immune effect of 4M2e- MAP,the BALB / c mice were immunized by NP_(67-74)- KLH and 4M2e- MAP after adding the complete freund's adjuvant and incomplete freund 's adjuvant to evaluate the immune effect. The weight change curve of mice after challenge showed that the weight change trend of mice immunized combinedly by NP_(67-74)- KLH and 4M2e- MAP was more smooth than that immunized by 4M2e- MAP,and the weight of mice in combined immunization group regained quickly. The results of fluorescent quantitation and lung tissue pathological slices showed that the compound polypeptide of NP_(67-74)- KLH with 4M2e- MAP could inhibit the virus replication in the lungs,reduce the pathological damage to lungs and get the effective protection during the course of resisting virus. This provided a good reference for the combined use of peptide vaccine against avian influenza research.
To explore the effect of T cell epitope peptide NP_(67-74)- KLH of NP protein on immune effect of 4M2e- MAP,the BALB / c mice were immunized by NP_(67-74)- KLH and 4M2e- MAP after adding the complete freund's adjuvant and incomplete freund 's adjuvant to evaluate the immune effect. The weight change curve of mice after challenge showed that the weight change trend of mice immunized combinedly by NP_(67-74)- KLH and 4M2e- MAP was more smooth than that immunized by 4M2e- MAP,and the weight of mice in combined immunization group regained quickly. The results of fluorescent quantitation and lung tissue pathological slices showed that the compound polypeptide of NP_(67-74)- KLH with 4M2e- MAP could inhibit the virus replication in the lungs,reduce the pathological damage to lungs and get the effective protection during the course of resisting virus. This provided a good reference for the combined use of peptide vaccine against avian influenza research.
引文
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[2]Claas E C J,Osterhaus A D M E,Rimmelzwaan G F,et al.Human influenza A H5N1 virus related to a highly pathogenic avian influenza virus[J].The Lancet,1998,351:472-477.
[3]Li Y,Shi J,Zhong G,et al.Continued evolution of H5N1 influenza virus in wild birds,domestic poultry,and humans in China from 2004 to 2009[J].J.Virol.,2010,84(17):8389-9397.
[4]Boon A C,de Mutsert G,Graus Y M,et al.The magnitude and specificity of influenza A virus-specific cytotoxic T-lymphocyte responses in humans is related to HLA-A and-B phenotype[J].J.Virol.,2002,76(2):582-90.
[5]Fan J,Liang X,Horton M S,et al.Preclinical study of influenza virus A M2 peptide conjugate vaccines in mice,ferrets,and rhesus monkeys[J].Vaccine,2004,22(23/24):2993-3003.
[6]Fiers W,De Filette M,Birkett A,et al.A“universal”human influenza A vaccine[J].Virus Res.,2004,103(1/2):173-176.
[7]Pertmer T M,Oran A E,Moser J M,et al.DNA vaccines for influenza virus:differential effects of maternal antibody on immune responses to hemagglutinin and nucleoprotein[J].J.Virol.,2000,74(17):7787-7793.
[8]Walzinger F,Ebner K,Lion T.Delection and monitoring of virus infections by real-time PCR[J].Molecular Aspects of Medicine,2006,27:254-298.