生物谷报道：抗体疗法的应用受到限制的部分原因是，生成完全属于人体的单克隆抗体非常困难。现在，通过识别人体免疫系统对某种特定病原体产生反应的时间窗口，Wrammert等人研究出一种有效生成高亲和力人体单克隆抗体的新方法。在接受针对流感病毒的增强免疫接种的个体中，血液中流感特定的IgG+抗体分泌细胞的数量在接种之后一周达到峰值，占全部外周B-细胞的大约6%。通过在这个时间窗口内收获B-细胞，就有可能产生超过50个人体单克隆抗体，它们 与三个具有高亲和力的目标流感病毒毒株结合在一起。整个过程所需时间不到一个月，要比传统单克隆生成方法快得多。该方法可应用于任何已有疫苗的传染病。
生物谷推荐英文原文：
Nature 453, 667-671 (29 May 2008) | doi:10.1038/nature06890; Received 16 October 2007; Accepted 4 March 2008; Published online 30 April 2008
Rapid cloning of high-affinity human monoclonal antibodies against influenza virus
Jens Wrammert1,10, Kenneth Smith3,10, Joe Miller1, William A. Langley1, Kenneth Kokko2, Christian Larsen2, Nai-Ying Zheng3, Israel Mays3, Lori Garman3, Christina Helms3, Judith James4,6,7, Gillian M. Air8, J. Donald Capra5,9, Rafi Ahmed1 & Patrick C. Wilson3,6,9
Emory Vaccine Center and Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA
Emory Transplant Center and Department of Surgery, Emory University School of Medicine, Atlanta, Georgia 30322, USA
Immunobiology and Cancer Research Program, The Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA
Arthritis and Immunology Research Program, The Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA
Molecular Immunogenetics Research Program, The Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA
Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA
Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA
Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA
These authors contributed equally to this work.
Correspondence to: Patrick C. Wilson3,6,9 Correspondence and requests for materials should be addressed to P.C.W. (Email: wilsonp@omrf.org).
Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14–21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine1. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.
