B cell response plays a critical role against SARS-CoV-2 infection. However, little is known about the diversity and frequency of the paired SARS-CoV-2 antigen-specific BCR repertoire after SARS-CoV-2 infection. Here, we performed single-cell RNA sequencing and VDJ sequencing using the memory and plasma B cells isolated from five convalescent COVID-19 patients, and analyzed the spectrum and transcriptional heterogeneity of antibody immune responses. Via linking BCR to antigen specificity through sequencing (LIBRA-seq), we identified a distinct activated memory B cell subgroup ( CD11c high CD95 high ) had a higher proportion of SARS-CoV-2 antigen-labeled cells compared with memory B cells. Our results revealed the diversity of paired BCR repertoire and the non-stochastic pairing of SARS-CoV-2 antigen-specific immunoglobulin heavy and light chains after SARS-CoV-2 infection. The public antibody clonotypes were shared by distinct convalescent individuals. Moreover, several antibodies isolated by LIBRA-seq showed high binding affinity against SARS-CoV-2 receptor-binding domain (RBD) or nucleoprotein (NP) via ELISA assay. Two RBD-reactive antibodies C14646P3S and C2767P3S isolated by LIBRA-seq exhibited high neutralizing activities against both pseudotyped and authentic SARS-CoV-2 viruses in vitro. Our study provides fundamental insights into B cell response following SARS-CoV-2 infection at the single-cell level.
【저자키워드】 Adaptive immunity, Infectious diseases, 【초록키워드】 SARS-CoV-2, antibody, SARS-COV-2 infection, Sequencing, memory B cells, SARS-CoV-2 virus, in vitro, heterogeneity, binding affinity, Single-cell RNA sequencing, memory, Antigen, specificity, Neutralizing activity, Immunoglobulin, RBD, immune responses, SARS-CoV-2 antigen, nucleoprotein, Spectrum, Critical, ELISA assay, memory B cell, BCR, convalescent COVID-19 patients, Frequency, Diversity, B cell response, domain, convalescent individuals, CD11c, pseudotyped, single-cell level, FIVE, Cell, memory B, transcriptional, plasma B cells, analyzed, performed, proportion, exhibited, provide, activated, plasma B cell, VDJ, Via, 【제목키워드】 SARS-CoV-2, immune profiling, Isolation, Rapid, Cell, memory B, convalescent COVID-19 patient,