Functionally Convergent B Cell Receptor Sequences in Transgenic Rats Expressing a Human B Cell Repertoire in Response to Tetanus Toxoid and Measles Antigens

AbstractThe identification and tracking of antigen-specific immunoglobulin (Ig) sequences within total Ig repertoires is central to high-throughput sequencing (HTS) studies of infections or vaccinations. In this context, public Ig sequences shared by different individuals exposed to the same antigen could be valuable markers for tracing back infections, measuring vaccine immunogenicity, and perhaps ultimately allow the reconstruction of the immunological history of an individual. Here, we immunized groups of transgenic rats expressing human Ig against tetanus toxoid (TT), Modified Vaccinia virus Ankara (MVA), measles virus hemagglutinin and fusion proteins expressed on MVA and the environmental carcinogen Benzo[a]Pyrene, coupled to TT. We showed that these antigens impose a selective pressure causing the Ig Heavy chain (IgH) repertoires of the rats to converge towards the expression of antibodies with highly similar IgH CDR3 amino acid sequences. We present a computational approach, similar to differential gene expression analysis, that selects for clusters of CDR3s with 80% similarity, significantly overrepresented within the different groups of immunized rats. These IgH clusters represent antigen-induced IgH signatures exhibiting stereotypic amino acid patterns including previously described TT and measles specific IgH sequences. Our data suggest, that with the presented methodology, transgenic Ig rats can be utilized as a model to identify antigen-induced, human IgH signatures to a variety of different antigens.

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Pre-B Cell Receptor Assesses the Quality of IgH Chains and Tunes the Pre-B Cell Repertoire by Delivering Differential Signals

The Journal of Immunology ◽

10.4049/jimmunol.177.4.2242 ◽

2006 ◽

Vol 177 (4) ◽

pp. 2242-2249 ◽

Cited By ~ 29

Author(s):

Yohei Kawano ◽

Soichiro Yoshikawa ◽

Yoshiyuki Minegishi ◽

Hajime Karasuyama

Keyword(s):

B Cell ◽

Cell Receptor ◽

B Cell Receptor ◽

Cell Repertoire ◽

B Cell Repertoire

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Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike

10.1101/2021.03.10.434840 ◽

2021 ◽

Author(s):

Pei Tong ◽

Avneesh Gautam ◽

Ian Windsor ◽

Meghan Travers ◽

Yuezhou Chen ◽

...

Keyword(s):

Monoclonal Antibodies ◽

B Cell ◽

Cell Receptor ◽

B Cell Receptor ◽

Cell Repertoire ◽

Memory B Cell ◽

Group Assignment ◽

Specific Memory ◽

B Cell Repertoire ◽

Protective Antibodies

ABSTRACTMemory B cell reserves can generate protective antibodies against repeated SARS-CoV-2 infections, but with an unknown reach from original infection to antigenically drifted variants. We charted memory B cell receptor-encoded monoclonal antibodies (mAbs) from 19 COVID-19 convalescent subjects against SARS-CoV-2 spike (S) and found 7 major mAb competition groups against epitopes recurrently targeted across individuals. Inclusion of published and newly determined structures of mAb-S complexes identified corresponding epitopic regions. Group assignment correlated with cross-CoV-reactivity breadth, neutralization potency, and convergent antibody signatures. mAbs that competed for binding the original S isolate bound differentially to S variants, suggesting the protective importance of otherwise-redundant recognition. The results furnish a global atlas of the S-specific memory B cell repertoire and illustrate properties conferring robustness against emerging SARS-CoV-2 variants.

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Transplanted Long-Term Cultured Pre-Bi Cells Expressing Calpastatin Are Resistant to B Cell Receptor–Induced Apoptosis

Journal of Experimental Medicine ◽

10.1084/jem.194.3.247 ◽

2001 ◽

Vol 194 (3) ◽

pp. 247-254 ◽

Cited By ~ 15

Author(s):

Antonio Ruiz-Vela ◽

Fernando Serrano ◽

Manuel A. González ◽

José Luis Abad ◽

Antonio Bernad ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Receptor ◽

B Cell Receptor ◽

Scid Mice ◽

Cell Repertoire ◽

Clonal Deletion ◽

B Cell Repertoire ◽

Induced Apoptosis

Long-term cultured pre-B cells are able to differentiate into immunoglobulin (Ig)M-positive B cells (IgM+ cells) when transplanted into severe combined immunodeficient (SCID) mice. Based on previous studies, here we report the development of a reconstitution assay in nonobese diabetic/SCID (NOD/SCID) mice using pre-B cells, which allows us to study the role of calpains (calcium-activated endopeptidases) during B cell development as well as in B cell clonal deletion. Using this model, we show that calpastatin (the natural inhibitor of calpains) inhibits B cell receptor–induced apoptosis in IgM+ cells derived from transplanted mice. We thus hypothesize an important function for calpain in sculpting the B cell repertoire.

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A public antibody class recognizes a novel S2 epitope exposed on open conformations of SARS-CoV-2 spike

10.1101/2021.12.01.470767 ◽

2021 ◽

Author(s):

Mathieu Claireaux ◽

Tom G Caniels ◽

Marlon de Gast ◽

Julianna Han ◽

Denise Guerra ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Receptor ◽

Memory B Cells ◽

B Cell Receptor ◽

Cell Repertoire ◽

Fold Enrichment ◽

B Cell Repertoire ◽

High Fraction ◽

Antibody Class

AbstractDelineating the origins and properties of antibodies elicited by SARS-CoV-2 infection and vaccination is critical for understanding their benefits and potential shortcomings. Therefore, we investigated the SARS-CoV-2 spike (S)-reactive B cell repertoire in unexposed individuals by flow cytometry and single-cell sequencing. We found that ∼82% of SARS-CoV-2 S-reactive B cells show a naive phenotype, which represents an unusually high fraction of total human naive B cells (∼0.1%). Approximately 10% of these naive S-reactive B cells shared an IGHV1-69/IGKV3-11 B cell receptor pairing, an enrichment of 18-fold compared to the complete naive repertoire. A proportion of memory B cells, comprising switched (∼0.05%) and unswitched B cells (∼0.04%), was also reactive with S and some of these cells were reactive with ADAMTS13, which is associated with thrombotic thrombocytopenia. Following SARS-CoV-2 infection, we report an average 37-fold enrichment of IGHV1-69/IGKV3-11 B cell receptor pairing in the S-reactive memory B cells compared to the unselected memory repertoire. This class of B cells targets a previously undefined non-neutralizing epitope on the S2 subunit that becomes exposed on S proteins used in approved vaccines when they transition away from the native pre-fusion state because of instability. These findings can help guide the improvement of SARS-CoV-2 vaccines.

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