Chan Zuckerberg Biohub, San Francisco, CA, United States.
Stanford ChEM-H and Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, United States.
Front Immunol. 2019 Jun 25;10:1452. doi: 10.3389/fimmu.2019.01452. eCollection 2019.
Phenotypic screening of antigen-specific antibodies in human blood is a common diagnostic test for infectious agents and a correlate of protection after vaccination. In addition to long-lived antibody secreting plasma cells residing in the bone marrow, memory B cells are a latent source of antigen-experienced, long-term immunity that can be found at low frequencies in circulating peripheral blood mononuclear cells (PBMCs). Assessing the genotype, clonal frequency, quality, and function of antibodies resulting from an individual's persistent memory B cell repertoire can help inform the success or failure of immune protection. Using polyclonal stimulation, we functionally expand the memory repertoire from PBMCs and clonally map monoclonal antibodies from this population. We show that combining deep sequencing of stimulated memory B cell repertoires with retrieving single antigen-specific cells is a promising approach in evaluating the latent, functional B cell memory in PBMCs.
在人体血液中进行抗原特异性抗体的表型筛选是一种常见的传染病诊断测试，也是疫苗接种后保护的相关指标。除了存在于骨髓中的长寿抗体分泌浆细胞外，记忆 B 细胞是一种潜在的抗原经验来源，具有长期免疫能力，在循环外周血单核细胞（PBMC）中以低频率存在。评估个体持续记忆 B 细胞库产生的抗体的基因型、克隆频率、质量和功能，有助于了解免疫保护的成败。我们使用多克隆刺激，从 PBMC 中功能扩展记忆库，并从该群体中克隆映射单克隆抗体。我们表明，将刺激后的记忆 B 细胞库的深度测序与检索单个抗原特异性细胞相结合，是评估 PBMC 中潜伏的、功能性 B 细胞记忆的一种很有前途的方法。
