Analysis of B cell receptor repertoires reveals key signatures of systemic B cell response after SARS-CoV-2 infection

A comprehensive study of the B cell response against SARS-CoV-2 could be significant for understanding the immune response and developing therapeutical antibodies and vaccines. To define the dynamics and characteristics of the antibody repertoire following SARS-CoV-2 infection, we analyzed the mRNA transcripts of immunoglobulin heavy chain (IgH) repertoires of 24 peripheral blood samples collected between 3 and 111 days after symptom onset from 10 COVID-19 patients. Massive clonal expansion of naïve B cells with limited somatic hypermutation (SHM) was observed in the second week after symptom onset. The proportion of low-SHM IgG clones strongly correlated with spike-specific IgG antibody titers, highlighting the significant activation of naïve B cells in response to a novel virus infection. The antibody isotype switching landscape showed a transient IgA surge in the first week after symptom onset, followed by a sustained IgG elevation that lasted for at least 3 months. SARS-CoV-2 infection elicited poly-germline reactive antibody responses. Interestingly, 17 different IGHV germline genes recombined with IGHJ6 showed significant clonal expansion. By comparing the IgH repertoires that we sequenced with the 774 reported SARS-CoV-2–reactive monoclonal antibodies (mAbs), 13 shared spike-specific IgH clusters were found. These shared spike-specific IgH clusters are derived from the same lineage of several recently published neutralizing mAbs, including CC12.1, CC12.3, C102, REGN10977, and 4A8. Furthermore, identical spike-specific IgH sequences were found in different COVID-19 patients, suggesting a highly convergent antibody response to SARS-CoV-2. Our analysis based on sequencing antibody repertoires from different individuals revealed key signatures of the systemic B cell response induced by SARS-CoV-2 infection. IMPORTANCE Although the canonical delineation of serum antibody responses following SARS-CoV-2 infection has been well established, the dynamics of antibody repertoire at the mRNA transcriptional level has not been well understood, especially the correlation between serum antibody titers and the antibody mRNA transcripts. In this study, we analyzed the IgH transcripts and characterized the B cell clonal expansion and differentiation, isotype switching, and somatic hypermutation in COVID-19 patients. This study provided insights at the repertoire level for the B cell response after SARS-CoV-2 infection.

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SARS-CoV-2 spike-specific memory B cells express higher levels of T-bet and FcRL5 after non-severe COVID-19 as compared to severe disease

PLoS ONE ◽

10.1371/journal.pone.0261656 ◽

2021 ◽

Vol 16 (12) ◽

pp. e0261656

Author(s):

Raphael A. Reyes ◽

Kathleen Clarke ◽

S. Jake Gonzales ◽

Angelene M. Cantwell ◽

Rolando Garza ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Symptom Onset ◽

Cell Response ◽

Severe Disease ◽

Memory B Cells ◽

Memory B Cell ◽

Specific Memory ◽

Specific Igg ◽

B Cell Response

SARS-CoV-2 infection elicits a robust B cell response, resulting in the generation of long-lived plasma cells and memory B cells. Here, we aimed to determine the effect of COVID-19 severity on the memory B cell response and characterize changes in the memory B cell compartment between recovery and five months post-symptom onset. Using high-parameter spectral flow cytometry, we analyzed the phenotype of memory B cells with reactivity against the SARS-CoV-2 spike protein or the spike receptor binding domain (RBD) in recovered individuals who had been hospitalized with non-severe (n = 8) or severe (n = 5) COVID-19. One month after symptom onset, a substantial proportion of spike-specific IgG+ B cells showed an activated phenotype. In individuals who experienced non-severe disease, spike-specific IgG+ B cells showed increased expression of markers associated with durable B cell memory, including T-bet and FcRL5, as compared to individuals who experienced severe disease. While the frequency of T-bet+ spike-specific IgG+ B cells differed between the two groups, these cells predominantly showed an activated switched memory B cell phenotype in both groups. Five months post-symptom onset, the majority of spike-specific memory B cells had a resting phenotype and the percentage of spike-specific T-bet+ IgG+ memory B cells decreased to baseline levels. Collectively, our results highlight subtle differences in the B cells response after non-severe and severe COVID-19 and suggest that the memory B cell response elicited during non-severe COVID-19 may be of higher quality than the response after severe disease.

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Prolonged evolution of the memory B cell response induced by a replicating adenovirus-influenza H5 vaccine

Science Immunology ◽

10.1126/sciimmunol.aau2710 ◽

2019 ◽

Vol 4 (34) ◽

pp. eaau2710 ◽

Cited By ~ 16

Author(s):

Kenta Matsuda ◽

Jinghe Huang ◽

Tongqing Zhou ◽

Zizhang Sheng ◽

Byong H. Kang ◽

...

Keyword(s):

B Cell ◽

Specific Antibody ◽

Cell Response ◽

Somatic Hypermutation ◽

Adenovirus Type ◽

Antigen Receptors ◽

Active Viral Replication ◽

Boost Vaccine ◽

Induced Immunity ◽

B Cell Response

Induction of an antibody response capable of recognizing highly diverse strains is a major obstacle to the development of vaccines for viruses such as HIV and influenza. Here, we report the dynamics of B cell expansion and evolution at the single-cell level after vaccination with a replication-competent adenovirus type 4 recombinant virus expressing influenza H5 hemagglutinin. Fluorescent H1 or H5 probes were used to quantitate and isolate peripheral blood B cells and their antigen receptors. We observed increases in H5-specific antibody somatic hypermutation and potency for several months beyond the period of active viral replication that was not detectable at the serum level. Individual broad and potent antibodies could be isolated, including one stem-specific antibody that is part of a new multidonor class. These results demonstrate prolonged evolution of the B cell response for months after vaccination and should be considered in efforts to evaluate or boost vaccine-induced immunity.

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Single-Cell Repertoire Analysis Demonstrates that Clonal Expansion Is a Prominent Feature of the B Cell Response in Multiple Sclerosis Cerebrospinal Fluid

The Journal of Immunology ◽

10.4049/jimmunol.171.5.2725 ◽

2003 ◽

Vol 171 (5) ◽

pp. 2725-2733 ◽

Cited By ~ 160

Author(s):

Gregory P. Owens ◽

Alanna M. Ritchie ◽

Mark P. Burgoon ◽

R. Anthony Williamson ◽

John R. Corboy ◽

...

Keyword(s):

Multiple Sclerosis ◽

Cerebrospinal Fluid ◽

B Cell ◽

Single Cell ◽

Cell Response ◽

Clonal Expansion ◽

Prominent Feature ◽

Cell Repertoire ◽

Repertoire Analysis ◽

B Cell Response

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Prolonged evolution of the human B cell response to SARS-CoV-2 infection

Science Immunology ◽

10.1126/sciimmunol.abg6916 ◽

2021 ◽

Vol 6 (56) ◽

pp. eabg6916

Author(s):

Mrunal Sakharkar ◽

C. Garrett Rappazzo ◽

Wendy F. Wieland-Alter ◽

Ching-Lin Hsieh ◽

Daniel Wrapp ◽

...

Keyword(s):

B Cell ◽

Cell Response ◽

De Novo ◽

Somatic Hypermutation ◽

Neutralizing Antibody ◽

Affinity Maturation ◽

Specific Igg ◽

Over Time ◽

Human B Cell ◽

B Cell Response

A comprehensive understanding of the kinetics and evolution of the human B cell response to SARS-CoV-2 infection will facilitate the development of next-generation vaccines and therapies. Here, we longitudinally profiled this response in mild and severe COVID-19 patients over a period of five months. Serum neutralizing antibody (nAb) responses waned rapidly but spike (S)-specific IgG+ memory B cells (MBCs) remained stable or increased over time. Analysis of 1,213 monoclonal antibodies (mAbs) isolated from S-specific MBCs revealed a primarily de novo response that displayed increased somatic hypermutation, binding affinity, and neutralization potency over time, providing evidence for prolonged antibody affinity maturation. B cell immunodominance hierarchies were similar across donor repertoires and remained relatively stable as the immune response progressed. Cross-reactive B cell populations, likely re-called from prior endemic beta-coronavirus exposures, comprised a small but stable fraction of the repertoires and did not contribute to the neutralizing response. The neutralizing antibody response was dominated by public clonotypes that displayed significantly reduced activity against SARS-CoV-2 variants emerging in Brazil and South Africa that harbor mutations at positions 501, 484 and 417 in the S protein. Overall, the results provide insight into the dynamics, durability, and functional properties of the human B cell response to SARS-CoV-2 infection and have implications for the design of immunogens that preferentially stimulate protective B cell responses.

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Lower IgG somatic hypermutation rates during acute dengue virus infection is compatible with a germinal center-independent B cell response

Genome Medicine ◽

10.1186/s13073-016-0276-1 ◽

2016 ◽

Vol 8 (1) ◽

Cited By ~ 19

Author(s):

Elizabeth Ernestina Godoy-Lozano ◽

Juan Téllez-Sosa ◽

Gilberto Sánchez-González ◽

Hugo Sámano-Sánchez ◽

Andrés Aguilar-Salgado ◽

...

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

B Cell ◽

Germinal Center ◽

Cell Response ◽

Somatic Hypermutation ◽

Dengue Virus Infection ◽

B Cell Response

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Abstract 416: Evaluation of Antibody Responses Toward Post-Translationally Modified and Unmodified Peptide Epitopes of Apolipoprotein A-I

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.36.suppl_1.416 ◽

2016 ◽

Vol 36 (suppl_1) ◽

Author(s):

David Henson ◽

Vincent J Venditto

Keyword(s):

B Cell ◽

Cell Response ◽

Polyclonal Antibodies ◽

Serum Markers ◽

Atherogenic Diet ◽

Antibody Responses ◽

Unmodified Peptide ◽

Peptide Epitopes ◽

B Cell Response

Antibody responses targeting ApoA-I have been documented in patients suffering from chronic inflammation associated with obesity, autoimmunity and cardiovascular disease (CVD) and correlate with a poor prognosis. Patients presenting with an acute myocardial infarction (AMI) have measurable IgG targeting the lecithin:cholesterol acyltransferase (LCAT) domain of ApoA-I and are shown to bind an epitope containing an oxidized methionine (Met-148(O)). Antibodies targeting ApoA-I are thought to be serum markers for CVD and not responsible for disease progression, but their exact role is poorly understood. We hypothesize that the B cell response to ApoA-I is nuanced with different isotypes of both protective and pathologic antibodies induced during the course of disease. To investigate this hypothesis, we synthesized a series of unmodified and modified peptides corresponding to the LCAT domain of mouse, monkey and human ApoA-I for ELISA and immunization studies. ELISA with commercially available goat polyclonal antibodies induced by immunization with human ApoA-I suggest that the LCAT domain possesses immunogenic properties and the antibodies are capable of binding to the unmodified and Met-148(O) peptides. Although affinity to the unmodified peptide appears to be higher by ELISA, pre-incubating the polyclonal antibodies with a solution of the Met-148(O) peptide significantly inhibits binding to the unmodified peptide. These data suggest that the oxidized peptide in solution may undergo a structural change that increases antibody affinity. Structural analysis of the peptides are ongoing. Serum analysis from patients presenting with an AMI and 30 days after cardiac event show a decline in IgG responses toward the Met-148(O)-modified peptide during that period. Analysis of monkey serum provides a unique comparison. Monkeys have a methionine to valine mutation at position 148, which results in a non-oxidizable residue, and instead show a decrease in IgG toward the unmodified sequence when on an atherogenic diet for 12 weeks. Correlating these ELISA assays with atherosclerosis observed in the patients and macaques along with ongoing in vivo immunization studies in mice will generate insight into the nuanced B cell response toward ApoA-I during CVD.

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SARS-CoV-2 spike-specific memory B cells express markers of durable immunity after non-severe COVID-19 but not after severe disease

10.1101/2021.09.24.461732 ◽

2021 ◽

Author(s):

Raphael Reyes ◽

Kathleen Clarke ◽

S. Jake Gonzales ◽

Angelene M. Cantwell ◽

Rolando Garza ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Symptom Onset ◽

Cell Response ◽

Severe Disease ◽

Memory B Cells ◽

Memory B Cell ◽

Specific Memory ◽

Specific Igg ◽

B Cell Response

SARS-CoV-2 infection elicits a robust B cell response, resulting in the generation of long-lived plasma cells and memory B cells. Here, we aimed to determine the effect of COVID-19 severity on the memory B cell response and characterize changes in the memory B cell compartment between recovery and five months post-symptom onset. Using high-parameter spectral flow cytometry, we analyzed the phenotype of memory B cells with reactivity against the SARS-CoV-2 spike protein or the spike receptor binding domain (RBD) in recovered individuals who had been hospitalized with non-severe (n=8) or severe (n=5) COVID-19. One month after symptom onset, a substantial proportion of spike-specific IgG+ B cells showed an activated phenotype. In individuals who experienced non-severe disease, spike-specific IgG+ B cells showed increased expression of markers associated with durable B cell memory, including T-bet, FcRL5, and CD11c, which was not observed after severe disease. Five months post-symptom onset, the majority of spike-specific memory B cells had a resting phenotype and the percentage of spike-specific T-bet+ IgG+ memory B cells decreased to baseline levels. Collectively, our results suggest that the memory B cell response elicited during non-severe COVID-19 may be of higher quality than the response after severe disease.

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