Phenotypic Characterization of Chinese Rhesus Macaque Plasmablasts for Cloning Antigen-Specific Monoclonal Antibodies

• Published in: Frontiers in immunology Vol. 10; p. 2426
• Main Authors: Zhang, Fan, Wang, Longyu, Niu, Xuefeng, Li, Jiashun, Luo, Jia, Feng, Yupeng, Yang, Yanjia, He, Ping, Fan, Wenxia, Liang, Renshan, Zheng, Zhiqiang, Pan, Weiqi, Li, Chufang, Tan, Yee Joo, Yu, Haijian, Chen, Ling, Li, Pingchao
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 11.10.2019
• Subjects: B cell; Chinese rhesus macaques; Immunology; influenza virus; monoclonal antibodies; plasmablast; vaccination; Chinese rhesus macaques; influenza virus; B cell; plasmablast; monoclonal antibodies; vaccination
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2019.02426

Rhesus macaques ( ) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that are highly similar to human-derived antibodies. Although antibody-secreting plasmablasts in humans are well-defined and can be easily isolated for mAb cloning, it remains unclear whether the same phenotypic markers could be applied for isolating antibody-secreting plasmablasts from Chinese rhesus macaques. In this study, we evaluated a series of cell surface and intracellular markers and identified the phenotypic markers of plasmablasts in Chinese rhesus macaques as CD3 CD14 CD56 CD19 CD27 CD20 CD80 HLA-DR CD95 . After influenza virus vaccination, the plasmablasts in peripheral blood mononuclear cells (PBMCs) increased transiently, peaked at day 4-7 after booster vaccination and returned to nearly undetectable levels by day 14. Antigen-specific enzyme-linked immunosorbent spot (ELISPOT) assays confirmed that the majority of the plasmablasts could produce influenza virus-specific antibodies. These plasmablasts showed transcriptional characteristics similar to those of human plasmablasts. Using single-cell PCR for immunoglobulin heavy and light chains, most mAbs cloned from the CD3 CD14 CD56 CD19 CD27 CD20 CD80 HLA-DR CD95 plasmablasts after vaccination exhibited specific binding to influenza virus. This study defined the phenotypic markers for isolating antibody-secreting plasmablasts from Chinese rhesus macaques, which has implications for efficient cloning of mAbs and for the evaluation of plasmablast response after vaccination or infection in Chinese rhesus macaques.