Functional characterization of monoclonal auto-antiidiotype antibodies isolated from the early B cell repertoire of BALB/c mice

AbstractEliciting broadly neutralizing antibodies (bNAbs) against the four dengue virus serotypes (DENV1-4) that are spreading into new territories is an important goal of vaccine design. To delineate bNAb targets, we characterized 28 monoclonal antibodies belonging to expanded and hypermutated clonal families identified by transcriptomic analysis of single plasmablasts from DENV-infected individuals. Among these, we identified two somatically related bNAbs that potently neutralized DENV1-4. Mutagenesis studies revealed that the major recognition determinants of these bNAbs are in E protein domain I, distinct from the only known class of human bNAbs against flaviviruses with a well-defined epitope. B cell repertoire analysis from acute-phase peripheral blood suggested a memory origin and divergent somatic hypermutation pathways for these bNAbs, and a limited number of mutations was sufficient for neutralizing activity. Our study suggests multiple B cell evolutionary pathways leading to DENV bNAbs targeting a novel epitope that can be exploited for vaccine design.

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B cell repertoire diversity in athymic mice.

Journal of Experimental Medicine ◽

10.1084/jem.151.3.761 ◽

1980 ◽

Vol 151 (3) ◽

pp. 761-766 ◽

Cited By ~ 8

Author(s):

M P Cancro ◽

N R Klinman

Keyword(s):

B Cell ◽

Pattern Analysis ◽

Athymic Mice ◽

Cell Repertoire ◽

Reactivity Pattern ◽

Influenza Hemagglutinin ◽

Immunoglobulin Allotype ◽

B Cell Repertoire ◽

Repertoire Diversity

The extent of B cell repertoire diversity among nu/nu BALB/c mice has been assessed and compared with that of normal BALB/c mice. This was accomplished through the characterization of monoclonal, influenza hemagglutinin-specific antibodies by reactivity pattern analysis. The results indicate that the repertoire of athymic mice is equivalent in diversity to that of normal mice. Moreover, because these responses were obtained in recipients that were histocompatible but distinct at immunoglobulin allotype loci, these findings indicate that a very diverse array of B cell clonotypes may be stimulated in the absence of allotype-identical T cells.

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Molecular characterization of the circulating anti-HIV-1 gp120-specific B cell repertoire using antibody phage display libraries generated from pre-selected HIV-1 gp120 binding PBLs

Journal of Immunological Methods ◽

10.1016/j.jim.2004.12.008 ◽

2005 ◽

Vol 297 (1-2) ◽

pp. 187-201 ◽

Cited By ~ 7

Author(s):

Klaus Koefoed ◽

Lauge Farnaes ◽

Meng Wang ◽

Arne Svejgaard ◽

Dennis R. Burton ◽

...

Keyword(s):

B Cell ◽

Cell Repertoire ◽

Gp120 Binding ◽

B Cell Repertoire ◽

Phage Display Libraries ◽

Antibody Phage ◽

Antibody Phage Display ◽

Anti Hiv ◽

Hiv 1

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Serum proteomics expands on high-affinity antibodies in immunized rabbits than deep B-cell repertoire sequencing alone

10.1101/833871 ◽

2019 ◽

Author(s):

Stefano R. Bonissone ◽

Thiago Lima ◽

Katherine Harris ◽

Laura Davison ◽

Brian Avanzino ◽

...

Keyword(s):

B Cell ◽

High Throughput ◽

Proteomic Analysis ◽

Intact Protein ◽

Serum Antibodies ◽

Cell Repertoire ◽

Serum Proteomics ◽

B Cell Repertoire ◽

Repertoire Sequencing

AbstractRabbits are a model for immunology studies, and monoclonal antibodies developed from rabbits have been sought after to empower immunoassays in a variety of applications. High-throughput characterization of circulating serum antibodies in response to specific antigens is highly impactful for both humoral immunology studies and antibody development. A combination of high throughput sequencing of antibody transcripts from B cells and proteomic analysis of serum antibodies, an approach referred to as immunoproteogenomics, is applied to profile the immune response of rabbits to β-galactosidase (Beta-gal) in both recombinant antigen and peptide antigen immunization formats. The use of intact protein antigen resulted in observing 56.3% more heavy chains CDR3s in serum than immunization with peptide antigens. Additionally, sampling peripheral blood mononuclear cells (PBMCs) for B-cell repertoire sequencing at different time points throughout the immunization was found to capture 47.8%-72.8% of total proteomically observed heavy chain CDR3s, and would serve well in replacing sequencing the B cell rich, but more difficult to access spleen or bone marrow compartments. Despite B-cell repertoire sequencing to depths of 2M to 10M reads, we found proteomic evidence supporting at least 10% of serum antibodies are still missed. Further improvements to proteomic analysis techniques would enable more precise characterization of antibodies circulating in serum and determine antibody protein sequences missed by repertoire sequencing.

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Characterization of T and B cell repertoire diversity in patients with RAG deficiency

Science Immunology ◽

10.1126/sciimmunol.aah6109 ◽

2016 ◽

Vol 1 (6) ◽

pp. eaah6109-eaah6109 ◽

Cited By ~ 42

Author(s):

Y. N. Lee ◽

F. Frugoni ◽

K. Dobbs ◽

I. Tirosh ◽

L. Du ◽

...

Keyword(s):

B Cell ◽

Cell Repertoire ◽

B Cell Repertoire ◽

Repertoire Diversity ◽

Rag Deficiency

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Review for "Templated Insertions at VD and DJ junctions Create Unique B‐Cell Receptors in the Healthy B‐Cell Repertoire"

10.1002/eji.202048828/v1/review2 ◽

2018 ◽

Keyword(s):

B Cell ◽

B Cell Receptors ◽

Cell Repertoire ◽

Cell Receptors ◽

B Cell Repertoire

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Selection of the expressed B cell repertoire by infusion of normal immunoglobulin G in a patient with autoimmune thyroiditis

European Journal of Immunology ◽

10.1002/eji.1830231133 ◽

1993 ◽

Vol 23 (11) ◽

pp. 2945-2950 ◽

Cited By ~ 40

Author(s):

Gilles Dietrich ◽

Francisco J. Varela ◽

Vincent Hurez ◽

Majida Bouanani ◽

Michel D. Kazatchkine

Keyword(s):

B Cell ◽

Immunoglobulin G ◽

Autoimmune Thyroiditis ◽

Cell Repertoire ◽

B Cell Repertoire ◽

Selection Of

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Monoclonal CD5+ and CD5- B-lymphocyte expansions are frequent in the peripheral blood of the elderly

Blood ◽

10.1182/blood-2003-09-3277 ◽

2004 ◽

Vol 103 (6) ◽

pp. 2337-2342 ◽

Cited By ~ 164

Author(s):

Paolo Ghia ◽

Giuseppina Prato ◽

Cristina Scielzo ◽

Stefania Stella ◽

Massimo Geuna ◽

...

Keyword(s):

B Cell ◽

Peripheral Blood ◽

B Lymphocyte ◽

The Elderly ◽

Lymphocytic Leukemia ◽

Pcr Analysis ◽

Cell Repertoire ◽

Healthy Elderly ◽

B Cell Repertoire ◽

Igh Genes

Abstract The responsiveness and diversity of peripheral B-cell repertoire decreases with age, possibly because of B-cell clonal expansions, as suggested by the incidence of serum monoclonal immunoglobulins and of monoclonal chronic lymphocytic leukemia (CLL)–like B lymphocytes in clinically silent adults. We phenotyped peripheral blood cells from 500 healthy subjects older than 65 years with no history or suspicion of malignancies and no evidence of lymphocytosis. In 19 cases (3.8%) a κ/λ ratio of more than 3:1 or less than 1:3 was found: 9 were CD5+, CD19+, CD23+, CD20low, CD79blow, sIglow (classic CLL-like phenotype); 3 were CD5+, CD19+, CD23+, CD20high, CD79blow, sIglow (atypical CLL-like), and 7 were CD5-, CD19+, CD20high, CD23-, CD79bbright, FMC7+, sIgbright (non–CLL-like). In 2 subjects, 2 phenotypically distinct unrelated clones were concomitantly evident. No cases were CD10+. Polymerase chain reaction (PCR) analysis demonstrated a monoclonal rearrangement of IgH genes in 15 of 19 cases. No bcl-1 or bcl-2 rearrangements were detected. Using a gating strategy based on CD20/CD5/CD79 expression, 13 additional CLL-like B-cell clones were identified (cumulative frequency of classic CLL-like: 5.5%). Thus, phenotypically heterogeneous monoclonal B-lymphocyte expansions are common among healthy elderly individuals and are not limited to classic CLL-like clones but may have the phenotypic features of different chronic lymphoproliferative disorders, involving also CD5- B cells.

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