Wilson Jason R, Tzeng Wen-Pin, Spesock April, Music Nedzad, Guo Zhu, Barrington Robert, Stevens James, Donis Ruben O, Katz Jacqueline M, York Ian A
Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
University of South Alabama, Mobile, AL, USA.
Virology. 2014 Jun;458-459:114-24. doi: 10.1016/j.virol.2014.04.011. Epub 2014 May 10.
We infected mice with the 2009 influenza A pandemic virus (H1N1pdm09), boosted with an inactivated vaccine, and cloned immunoglobulins (Igs) from HA-specific B cells. Based on the redundancy in germline gene utilization, we inferred that between 72-130 unique IgH VDJ and 35 different IgL VJ combinations comprised the anti-HA recall response. The IgH VH1 and IgL VK14 variable gene families were employed most frequently. A representative panel of antibodies were cloned and expressed to confirm reactivity with H1N1pdm09 HA. The majority of the recombinant antibodies were of high avidity and capable of inhibiting H1N1pdm09 hemagglutination. Three of these antibodies were subtype-specific cross-reactive, binding to the HA of A/South Carolina/1/1918(H1N1), and one further reacted with A/swine/Iowa/15/1930(H1N1). These results help to define the genetic diversity of the influenza anti-HA antibody repertoire profile induced following infection and vaccination, which may facilitate the development of influenza vaccines that are more protective and broadly neutralizing.
Protection against influenza viruses is mediated mainly by antibodies, and in most cases this antibody response is narrow, only providing protection against closely related viruses. In spite of this limited range of protection, recent findings indicate that individuals immune to one influenza virus may contain antibodies (generally a minority of the overall response) that are more broadly reactive. These findings have raised the possibility that influenza vaccines could induce a more broadly protective response, reducing the need for frequent vaccine strain changes. However, interpretation of these observations is hampered by the lack of quantitative characterization of the antibody repertoire. In this study, we used single-cell cloning of influenza HA-specific B cells to assess the diversity and nature of the antibody response to influenza hemagglutinin in mice. Our findings help to put bounds on the diversity of the anti-hemagglutinin antibody response, as well as characterizing the cross-reactivity, affinity, and molecular nature of the antibody response.
我们用2009年甲型大流行性流感病毒（H1N1pdm09）感染小鼠，用灭活疫苗加强免疫，然后从HA特异性B细胞中克隆免疫球蛋白（Ig）。基于种系基因利用的冗余性，我们推断72 - 130个独特的IgH VDJ和35种不同的IgL VJ组合构成了抗HA回忆反应。IgH VH1和IgL VK14可变基因家族的使用最为频繁。克隆并表达了一组代表性抗体以确认其与H1N1pdm09 HA的反应性。大多数重组抗体具有高亲和力，能够抑制H1N1pdm09血凝。其中三种抗体具有亚型特异性交叉反应性，可与A/南卡罗来纳/1/1918（H1N1）的HA结合，另一种与A/猪/爱荷华/15/1930（H1N1）反应。这些结果有助于确定感染和接种疫苗后诱导的流感抗HA抗体库谱的遗传多样性，这可能有助于开发更具保护性和广泛中和作用的流感疫苗。
针对流感病毒的保护主要由抗体介导，在大多数情况下，这种抗体反应具有局限性，仅能针对密切相关的病毒提供保护。尽管保护范围有限，但最近的研究结果表明，对一种流感病毒免疫的个体可能含有反应性更广泛的抗体（通常占总体反应的少数）。这些发现增加了流感疫苗可以诱导更广泛保护反应的可能性，从而减少对频繁更换疫苗毒株的需求。然而，由于缺乏抗体库的定量特征描述，这些观察结果的解释受到了阻碍。在本研究中，我们使用流感HA特异性B细胞的单细胞克隆来评估小鼠对流感血凝素抗体反应的多样性和性质。我们的研究结果有助于界定抗血凝素抗体反应的多样性，以及表征抗体反应的交叉反应性、亲和力和分子性质。
