甲型H7N9流感病毒疫苗经不同途径免疫小鼠长期保护效果的比较
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摘要
目的比较甲型H7N9流感全病毒灭活疫苗和裂解疫苗经3种免疫途径免疫小鼠的长期保护效果。方法采用不同剂量(0. 15和1. 5μg)的甲型H7N9流感全病毒灭活疫苗及裂解疫苗单独免疫或辅以Al(OH)_3佐剂共同免疫,分别经腹腔、肌肉及皮下注射免疫小鼠1次,免疫后不同时间点(3～360 d)收集血清,通过ELISA法检测血清IgG抗体水平。免疫后365 d,用含10×LD_(50)致死量的甲型H7N9流感病毒同源鼠肺适应株A/Shanghai/2/2013(H7N9)经滴鼻攻毒,20μL/只,攻毒3 d,检测小鼠肺洗液的病毒滴度,攻毒21 d,观察小鼠存活和体重丢失情况。结果甲型H7N9流感全病毒灭活疫苗和裂解疫苗接种1次均可诱导小鼠产生较高水平的血清IgG抗体,全病毒灭活疫苗比裂解疫苗IgG抗体滴度峰值高2～8倍,且Al(OH)_3佐剂可有效提高两种疫苗诱导的血清IgG抗体水平。免疫后365 d,全病毒灭活疫苗及含佐剂裂解疫苗免疫均可保护小鼠抵抗同源流感病毒的致死量攻击,且Al(OH)_3佐剂可有效提高H7N9裂解疫苗的保护效果。3种免疫途径中腹腔注射和肌肉注射较皮下注射产生的IgG抗体维持时间更长,免疫剂量相同时,肌肉免疫途径对小鼠的保护效果略优于腹腔和皮下免疫途径。结论 3种途径1次免疫甲型H7N9流感全病毒灭活疫苗或辅以Al(OH)_3佐剂的裂解疫苗,在小鼠模型中可提供至少365 d的保护,腹腔和肌肉免疫途径血清抗体持续时间较皮下途径更长,肌肉免疫途径保护效果略优于另两种途径。
Objective To compare the long-term protective effects of inactivated whole and split virion H7N9 influenza vaccines in mice by three immunization routes. Methods Female BALB/c mice were immunized once with inactivated whole or split virion H7N9 influenza vaccines with or without aluminum hydroxide adjuvant at dosages of 0. 15 and 1. 5 μg,by intraperitoneal(i.p.),intramuscular(i.m.)or subcutaneous(s.c.)route. Serum samples were collected at different time points(3 ～ 360 d)after immunization and determined for the titers of IgG antibody by ELISA. The mice were challenged with homologous virus strain A/Shanghai/2/2013(H7N9)at 10 × LD_(50) by intranasal drip 365 d after immunization,20 μL for each. The virus titer in bronchoalveolar wash was determined 3 d,while the survival and weight loss were monitored 21 d,after the challenge. Results A single dose of inactivated whole and split virion H7N9 influenza vaccine induced high serum IgG antibody level in mice. The peak of IgG antibody level of mice induced with whole virion vaccine was 2 ～ 8 times higher than that with split virion vaccine,while the aluminum hydroxide adjuvant significantly enhanced the IgG antibody levels induced by the two kinds of vaccine. The whole virion vaccine and the split virion vaccine containing aluminum hydroxide adjuvant protected the mice against a lethal challenge with homologous influenza virus 365 d after immunization,while the aluminum hydroxide adjuvant significantly enhanced the protective effect of H7N9 split vaccine. The antibody levels induced by i.p. and i.m. routes were more persistent than that by s.c. route. However,at the same dosage,the protective effect of vaccine by i.m. route was slightly superior to those by i.p. and s.c. routes. Conclusion The protection lasting for at least 365 d were induced by a single dose of inactivated whole virion H7N9 influenza vaccine or the split viron vaccine containing aluminum hydroxide adjuvant by three immunization routes. The antibody levels induced by i.p. and i.m. routes were more persistent,while the protective effect by i.m. route was slightly superior to those by the other two routes.
Objective To compare the long-term protective effects of inactivated whole and split virion H7N9 influenza vaccines in mice by three immunization routes. Methods Female BALB/c mice were immunized once with inactivated whole or split virion H7N9 influenza vaccines with or without aluminum hydroxide adjuvant at dosages of 0. 15 and 1. 5 μg,by intraperitoneal(i.p.),intramuscular(i.m.)or subcutaneous(s.c.)route. Serum samples were collected at different time points(3 ～ 360 d)after immunization and determined for the titers of IgG antibody by ELISA. The mice were challenged with homologous virus strain A/Shanghai/2/2013(H7N9)at 10 × LD_(50) by intranasal drip 365 d after immunization,20 μL for each. The virus titer in bronchoalveolar wash was determined 3 d,while the survival and weight loss were monitored 21 d,after the challenge. Results A single dose of inactivated whole and split virion H7N9 influenza vaccine induced high serum IgG antibody level in mice. The peak of IgG antibody level of mice induced with whole virion vaccine was 2 ～ 8 times higher than that with split virion vaccine,while the aluminum hydroxide adjuvant significantly enhanced the IgG antibody levels induced by the two kinds of vaccine. The whole virion vaccine and the split virion vaccine containing aluminum hydroxide adjuvant protected the mice against a lethal challenge with homologous influenza virus 365 d after immunization,while the aluminum hydroxide adjuvant significantly enhanced the protective effect of H7N9 split vaccine. The antibody levels induced by i.p. and i.m. routes were more persistent than that by s.c. route. However,at the same dosage,the protective effect of vaccine by i.m. route was slightly superior to those by i.p. and s.c. routes. Conclusion The protection lasting for at least 365 d were induced by a single dose of inactivated whole virion H7N9 influenza vaccine or the split viron vaccine containing aluminum hydroxide adjuvant by three immunization routes. The antibody levels induced by i.p. and i.m. routes were more persistent,while the protective effect by i.m. route was slightly superior to those by the other two routes.
引文
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[13]European Pharmacopoeia 8.0[S].Strasbourg:EDQM,2008:1664.
[14]HE P,ZOU Y,HU Z.Advances in aluminum hydroxide-based adjuvant research and its mechanism[J].Hum Vaccin Immunother,2015,11(2):477-488.
[15]GHERARDI R K,AOUIZERATE J,CADUSSEAUJ,et al.Aluminum adjuvants of vaccines injected into the muscle:normal fate,pathology and associated disease[J].Morphologie,2016,100(329):85-94.
[16]MCKEE A S,MARRACK P.Old and new adjuvants[J].Curr Opin Immunol,2017,47:44-51.
[17]AJANA F,SANA C,CAULIN E,Are there differences in immunogenicity and safety of vaccines according to the injection method[J].Med Mal Infect,2008,38(12):648-657.
[18]SEALE J R C.Importance of injecting vaccines into muscle.Parenteral vaccines must be given subcutaneously in patients with congenital bleeding disorders[J].BMJ,2001,322(7282):364.
[19]COOK I F,BARR I,HARTEL G,et al.Reactogenicity and immunogenicity of an inactivated influenza vaccine administered by intramuscular or subcutaneous injection in elderly adults[J].Vaccine,2006,24(13):2395-2402.
[2]WHO.Influenza at the human-animal iterface,26 January 26-2March 2018[EB/OL].(2018-03-20)[2018-03-20]http://www.who.int/influenza/hu man_animal_interface/Influenza_Summary_IRA_HA_interface_02_03_2018.pdf?ua=1.
[3]WANG X,JIANG H,WU P,et al.Epidemiology of avian influenza A H7N9 virus in human beings across five epidemics in mainland China,2013-17:an epidemiological study of laboratory-confirmed case series[J].Lancet Infect Dis,17(8):822-832.
[4]WANG X,et al.Assessment of human-to-human transmissibility of avian influenza A(H7N9)virus across five waves by analyzing clusters of case-patients in mainland China,2013-2017[J].Clin Infect Dis,2018,doi:10.1093/cid/ciy541.
[5]JACKSON L A,CAMPBELL J D,FREY S E,et al.Effect of varying doses of a monovalent H7N9 influenza vaccine with and without AS03 and MF59 adjuvants on immune response:a randomized clinical trial[J].JAMA,2015,314(3):237-246.
[6]MADAN A,SEGALL N,FERGUSON M,et al.Immunogenicity and safety of an AS03-adjuvanted H7N9 pandemic influenza vaccine in a randomized trial in healthy adults[J].J Infect Dis,2016,214(11):1717-1727.
[7]WU U I,HSIEH S M,LEE W S,et al.Safety and immunogenicity of an inactivated cell culture-derived H7N9 influenza vaccine in healthy adults:A phase I/II,prospective,randomized,open-label trial[J].Vaccine,2017,35(33):4099-4104.
[8]RUDENKO L,KISELEVA I,NAYKHIN A N,et al.Assessment of human immune responses to H7 avian influenza virus of pandemic potential:results from a placebo-controlled,randomized double-blind phase I study of live attenuated H7N3 influenza vaccine[J].PLoS One,2014,9(2):e87962.
[9]MADAN A,FERGUSON M,RHEAULT P,et al.Immunogenicity and safety of an AS03-adjuvanted H7N1 vaccine in adults 65 years of age and older:A phase II,observer-blind,randomized,controlled trial[J].Vaccine,2017,35(15):1865-1872.
[10]CHUNG K Y,COYLE E M,JANI D,et al.ISCOMATRIXTMadjuvant promotes epitope spreading and antibody affinity maturation of influenza A H7N9 virus like particle vaccine that correlate with virus neutralization in humans[J].Vaccine,2015,33(32):3953-3962.
[11]ZHENG D,GAO F,ZHAO C,et al.Comparative effectiveness of H7N9 vaccines in healthy individuals[J].Hum Vaccin Immunother,2018,1-11.
[12]LIN Y J,SHIH Y J,CHEN C H,et al.Aluminum salts as an adjuvant for pre-pandemic influenza vaccines:a meta-analysis[J].Scientific Reports,2018,30(1):8.
[13]European Pharmacopoeia 8.0[S].Strasbourg:EDQM,2008:1664.
[14]HE P,ZOU Y,HU Z.Advances in aluminum hydroxide-based adjuvant research and its mechanism[J].Hum Vaccin Immunother,2015,11(2):477-488.
[15]GHERARDI R K,AOUIZERATE J,CADUSSEAUJ,et al.Aluminum adjuvants of vaccines injected into the muscle:normal fate,pathology and associated disease[J].Morphologie,2016,100(329):85-94.
[16]MCKEE A S,MARRACK P.Old and new adjuvants[J].Curr Opin Immunol,2017,47:44-51.
[17]AJANA F,SANA C,CAULIN E,Are there differences in immunogenicity and safety of vaccines according to the injection method[J].Med Mal Infect,2008,38(12):648-657.
[18]SEALE J R C.Importance of injecting vaccines into muscle.Parenteral vaccines must be given subcutaneously in patients with congenital bleeding disorders[J].BMJ,2001,322(7282):364.
[19]COOK I F,BARR I,HARTEL G,et al.Reactogenicity and immunogenicity of an inactivated influenza vaccine administered by intramuscular or subcutaneous injection in elderly adults[J].Vaccine,2006,24(13):2395-2402.