Kamal Ram P, Blanchfield Kristy, Belser Jessica A, Music Nedzad, Tzeng Wen-Pin, Holiday Crystal, Burroughs Ashley, Sun Xiangjie, Maines Taronna R, Levine Min Z, York Ian A
Battelle Memorial Institute, Atlanta, Georgia, USA
Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
J Virol. 2017 Sep 27;91(20). doi: 10.1128/JVI.01202-17. Print 2017 Oct 15.
Avian influenza viruses of the H7 hemagglutinin (HA) subtype present a significant public health threat, as evidenced by the ongoing outbreak of human A(H7N9) infections in China. When evaluated by hemagglutination inhibition (HI) and microneutralization (MN) assays, H7 viruses and vaccines are found to induce lower level of neutralizing antibodies (nAb) than do their seasonal counterparts, making it difficult to develop and evaluate prepandemic vaccines. We have previously shown that purified recombinant H7 HA appear to be poorly immunogenic in that they induce low levels of HI and MN antibodies. In this study, we immunized mice with whole inactivated reverse genetics reassortant (RG) viruses expressing HA and neuraminidase (NA) from 3 different H7 viruses [A/Shanghai/2/2013(H7N9), A/Netherlands/219/2003(H7N7), and A/New York/107/2003(H7N2)] or with human A(H1N1)pdm09 (A/California/07/2009-like) or A(H3N2) (A/Perth16/2009) viruses. Mice produced equivalent titers of antibodies to all viruses as measured by enzyme-linked immunosorbent assay (ELISA). However, the antibody titers induced by H7 viruses were significantly lower when measured by HI and MN assays. Despite inducing very low levels of nAb, H7 vaccines conferred complete protection against homologous virus challenge in mice, and the serum antibodies directed against the HA head region were capable of mediating protection. The apparently low immunogenicity associated with H7 viruses and vaccines may be at least partly related to measuring antibody titers with the traditional HI and MN assays, which may not provide a true measure of protective immunity associated with H7 immunization. This study underscores the need for development of additional correlates of protection for prepandemic vaccines. H7 avian influenza viruses present a serious risk to human health. Preparedness efforts include development of prepandemic vaccines. For seasonal influenza viruses, protection is correlated with antibody titers measured by hemagglutination inhibition (HI) and virus microneutralization (MN) assays. Since H7 vaccines typically induce low titers in HI and MN assays, they have been considered to be poorly immunogenic. We show that in mice H7 whole inactivated virus vaccines (WIVs) were as immunogenic as seasonal WIVs, as they induced similar levels of overall serum antibodies. However, a larger fraction of the antibodies induced by H7 WIV was nonneutralizing Nevertheless, the H7 WIV completely protected mice against homologous viral challenge, and antibodies directed against the HA head were the major contributor toward immune protection. Vaccines against H7 avian influenza viruses may be more effective than HI and virus neutralization assays suggest, and such vaccines may need other methods for evaluation.
H7血凝素(HA)亚型禽流感病毒对公众健康构成重大威胁,中国持续爆发的人感染A(H7N9)疫情就是明证。通过血凝抑制(HI)和微量中和(MN)试验评估发现,H7病毒和疫苗诱导产生的中和抗体(nAb)水平低于季节性流感病毒,这使得大流行前疫苗的研发和评估变得困难。我们之前已经表明,纯化的重组H7 HA免疫原性似乎较差,因为它们诱导产生的HI和MN抗体水平较低。在本研究中,我们用表达来自3种不同H7病毒[A/上海/2/2013(H7N9)、A/荷兰/219/2003(H7N7)和A/纽约/107/2003(H7N2)]的HA和神经氨酸酶(NA)的全灭活反向遗传学重配(RG)病毒,或用人A(H1N1)pdm09(A/加利福尼亚/07/2009样)或A(H3N2)(A/珀斯16/2009)病毒免疫小鼠。通过酶联免疫吸附测定(ELISA)测量,小鼠对所有病毒产生的抗体滴度相当。然而,通过HI和MN试验测量时,H7病毒诱导产生的抗体滴度显著较低。尽管诱导产生的nAb水平非常低,但H7疫苗在小鼠中对同源病毒攻击提供了完全保护,并且针对HA头部区域的血清抗体能够介导保护作用。与H7病毒和疫苗相关的明显低免疫原性可能至少部分与用传统的HI和MN试验测量抗体滴度有关,这可能无法真正衡量与H7免疫相关的保护性免疫。本研究强调了为大流行前疫苗开发其他保护相关性指标的必要性。H7禽流感病毒对人类健康构成严重风险。防范措施包括研发大流行前疫苗。对于季节性流感病毒,保护作用与通过血凝抑制(HI)和病毒微量中和(MN)试验测量的抗体滴度相关。由于H7疫苗在HI和MN试验中通常诱导产生低滴度,它们一直被认为免疫原性较差。我们表明,在小鼠中,H7全灭活病毒疫苗(WIV)与季节性WIV具有相同的免疫原性,因为它们诱导产生的总体血清抗体水平相似。然而,H7 WIV诱导产生的抗体中较大一部分是非中和性的。尽管如此,H7 WIV完全保护小鼠免受同源病毒攻击,并且针对HA头部的抗体是免疫保护的主要贡献者。针对H7禽流感病毒的疫苗可能比HI和病毒中和试验显示的更有效,并且此类疫苗可能需要其他评估方法。