一种鉴定人血液中HIV gp140结合记忆B细胞的方法。
A method for identification of HIV gp140 binding memory B cells in human blood.
作者信息
Scheid Johannes F, Mouquet Hugo, Feldhahn Niklas, Walker Bruce D, Pereyra Florencia, Cutrell Emily, Seaman Michael S, Mascola John R, Wyatt Richard T, Wardemann Hedda, Nussenzweig Michel C
机构信息
Laboratory of Molecular Immunology, The Rockefeller University, New York, New York 10065, USA.
出版信息
J Immunol Methods. 2009 Apr 15;343(2):65-7. doi: 10.1016/j.jim.2008.11.012. Epub 2008 Dec 25.
Antibodies to HIV are potentially important reagents for basic and clinical studies. Historically, these reagents have been produced by random cloning of heavy and light chains in phage display libraries [Burton, D.R., Barbas, C.F. III, Persson, M.A.A., Koenig, S., Chanock, R.M., and Lerner, R.A., (1991), A large array of human monoclonal antibodies to type 1 immunodeficiency virus from combinatorial libraries of asymptomatic seropositive individuals. Proc. Natl. Acad. Sci. U. S. A. 88, 10134-10137.] and electrofusion techniques [Buchacher, A., Predl, R., Tauer, C., Purtscher, M., Gruber, G., Heider, R., Steindl, F., Trkola, A., Jungbauer, A., and Katinger, H., (1992), Human monoclonal antibodies against gp41 and gp120 as potential agent for passive immunization. Vaccines 92, 191-195]. Here we describe a method to identify and potentially enrich human memory B cells from HIV infected patients that show serum titers of neutralizing antibodies. When biotinylated gp140 is used to stain peripheral blood mononuclear cells it identifies a distinct population of gp140 binding B cells by flow cytometry.
抗HIV抗体是基础研究和临床研究中潜在的重要试剂。从历史上看，这些试剂是通过在噬菌体展示文库中随机克隆重链和轻链来生产的[伯顿，D.R.，巴尔巴斯，C.F.三世，佩尔松，M.A.A.，凯尼格，S.，查诺克，R.M.，和勒纳，R.A.，（1991年），来自无症状血清阳性个体组合文库的大量针对1型免疫缺陷病毒的人单克隆抗体。美国国家科学院院刊88，10134 - 10137。]以及电融合技术[布哈彻，A.，普雷德尔，R.，陶尔，C.，普尔切尔，M.，格鲁伯，G.，海德，R.，施泰因德尔，F.，特科拉，A.，容鲍尔，A.，和卡廷格，H.，（1992年）, 抗gp41和gp120的人单克隆抗体作为被动免疫的潜在试剂。疫苗92，191 - 195]。在这里，我们描述了一种从显示中和抗体血清滴度的HIV感染患者中鉴定并可能富集人记忆B细胞的方法。当用生物素化的gp140对外周血单核细胞进行染色时，通过流式细胞术可识别出一个独特的gp140结合B细胞群体。
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