Production and Characterization of Human Monoclonal Antibodies from the Cells of A(H1N1)pdm2009 Influenza Virus Infected Indian Donors

Latika Saxena, Madhu Khanna
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Abstract

Analysis of human monoclonal antibodies (mAbs) developed from influenza infected donors have enormously contributed to the identification of neutralization sensitive epitopes of influenza virus. The HA protein is a crucial target of neutralizing antibodies and at monoclonal level only Abs binding to HA have been able to neutralize the virus. In this study, eight A (H1N1)pdm 2009 seropositive patients within the age range of 20-50 years (median = 36 years) were recruited. Two anti-HA mAbs secreting stable clones, 2D8 and 2F12 were established under optimized conditions from the peripheral blood mononuclear cells (PBMCs) of the volunteers. These antibodies efficiently neutralized the homologous laboratory isolated strain of the pandemic virus as well as the reference strain. Our study suggests that the anti-HA antibodies derived from infected Indian patients display neutralization potential against the A(H1N1)pdm 2009 virus. This is the first ever study of generation of mAbs against the pandemic influenza virus involving the immune repertoire if Indian patients. Molecular characterization of the target regions will help in identifying potential immunogens in the Indian pandemic isolates and confer protective immunity against this virus.

从感染甲型H1N1流感病毒的印度供体细胞中提取人单克隆抗体的研究
从流感感染供体获得的人单克隆抗体(mab)的分析极大地促进了流感病毒中和敏感表位的鉴定。HA蛋白是中和抗体的关键靶点,在单克隆水平上,只有与HA结合的抗体能够中和病毒。本研究招募了8例年龄在20-50岁(中位年龄36岁)的甲型H1N1 pdm 2009血清阳性患者。以志愿者外周血单个核细胞(PBMCs)为样本,在优化条件下建立稳定克隆的抗ha单克隆抗体2D8和2F12。这些抗体有效地中和了大流行病毒的同源实验室分离株以及参考株。我们的研究表明,来自受感染印度患者的抗ha抗体对甲型H1N1 pdm 2009病毒具有中和潜力。这是有史以来第一次研究针对大流行性流感病毒的单克隆抗体,涉及印度患者的免疫库。目标区域的分子特征将有助于确定印度大流行分离株中潜在的免疫原,并赋予对该病毒的保护性免疫。
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