Somatic diversification of immunoglobulin heavy chain VDJ genes: Evidence for somatic gene conversion in rabbits

Becker, Robert S.; Knight, Katherine L.

doi:10.1016/0092-8674(90)90502-6

Cited by 246 publications

(125 citation statements)

References 30 publications

Supporting

Mentioning

123

Contrasting

Unclassified

Order By: Relevance

“…S3). Because gene conversion is an important mechanism for Ig diversification in the rabbit (22), it cannot be ascertained whether the highly mutated V gene population resulted from activation-induced deaminase (AID) mutagenesis or from recombination events with V pseudogenes.…”

Section: Resultsmentioning

confidence: 99%

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Wine

Boutz

Lavinder

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

129

156

View full text Add to dashboard Cite

We have developed and validated a methodology for determining the antibody composition of the polyclonal serum response after immunization. Pepsin-digested serum IgGs were subjected to standard antigen-affinity chromatography, and resulting elution, wash, and flow-through fractions were analyzed by bottom-up, liquid chromatography-high-resolution tandem mass spectrometry. Identification of individual monoclonal antibodies required the generation of a database of IgG variable gene (V-gene) sequences constructed by NextGen sequencing of mature B cells. Antibody V-gene sequences are characterized by short complementarity determining regions (CDRs) of high diversity adjacent to framework regions shared across thousands of IgGs, greatly complicating the identification of antigen-specific IgGs from proteomically observed peptides. By mapping peptides marking unique V H CDRH3 sequences, we identified a set of V-genes heavily enriched in the affinity chromatography elution, constituting the serum polyclonal response. After booster immunization in a rabbit, we find that the antigenspecific serum immune response is oligoclonal, comprising antibodies encoding 34 different CDRH3s that group into 30 distinct antibody V H clonotypes. Of these 34 CDRH3s, 12 account for ∼60% of the antigen-specific CDRH3 peptide mass spectral counts. For comparison, antibodies with 18 different CDRH3s (12 clonotypes) were represented in the antigen-specific IgG fraction from an unimmunized rabbit that fortuitously displayed a moderate titer for BSA. Proteomically identified antibodies were synthesized and shown to display subnanomolar affinities. The ability to deconvolute the polyclonal serum response is likely to be of key importance for analyzing antibody responses after vaccination and for more completely understanding adaptive immune responses in health and disease.antibody proteomics | antibody repertoire | serum immunoprofiling | B-cell response | humoral response T he first Nobel Prize in Medicine was awarded to Emil von Behring, who in collaboration with Kitasato Shibasaburo and Paul Ehrlich discovered serum antitoxins (1, 2). Remarkably, after more than 100 y of intense research in immunology, little is known about the clonality, relative concentrations, and binding properties of the monoclonal antibodies that constitute the antigen-specific Ig pool in serum.At steady state, circulating antibodies are produced by terminally differentiated B lymphocytes (plasma cells) within the bone marrow, and thus cannot be accessed in living individuals (3). Although recent single B-cell cloning methods (4, 5) have led to the identification of peripheral antigen-specific B memory and/or antibody-secreting cells (plasmablasts), it is generally unknown whether the Igs encoded by peripheral blood B cells correspond to the antibodies present in circulation and especially whether they are present at physiologically relevant levels (i.e., at serum concentrations above K D corresponding to >1 μg/mL for an average affinity of individual antibodies of 5 nM)...

show abstract

Section: Resultsmentioning

confidence: 99%

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Wine

Boutz

Lavinder

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

129

156

View full text Add to dashboard Cite

show abstract

“…For instance, the best VLRA HEL-binding clone (R5.1) had a K D of 124 pM, approaching the 100 pM affinity ceiling of Ig-based antibodies produced during mammalian immune responses (1,24). Future studies will address the question of whether these VLRA variants arose as part of the primary repertoire, as in sheep and rabbits (25,26), or as a consequence of antigen-driven affinity maturation of antibody responses, as in all jawed vertebrates from shark to man (2). In conclusion, the powerful YSD platform described herein has allowed us to explore the role of VLR in lamprey immunity by screening large libraries for specific antigen-binding VLR clones.…”

Section: <-----Lrrnt -----><------Lrr1 ------><----Lrrve ------><---Cp ---><-----------------Lrrct ----------------mentioning

confidence: 99%

High-affinity lamprey VLRA and VLRB monoclonal antibodies

Tasumi

Velikovsky

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

104

138

View full text Add to dashboard Cite

show abstract

“…It has now become apparent that gene conversion is not restricted to birds, but also occurs in rabbits (Becker & Knight, 1990) and cattle, sheep and horses (reviewed in Butler, 1998), although these species do not solely use gene conversion.…”

Section: The Chicken's Contribution To Immunology As a Wholementioning

confidence: 99%

The long view: a bright past, a brighter future? Forty years of chicken immunology pre- and post-genome

Kaiser

2012

Avian Pathology

View full text Add to dashboard Cite

Somatic diversification of immunoglobulin heavy chain VDJ genes: Evidence for somatic gene conversion in rabbits

Cited by 246 publications

References 30 publications

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

High-affinity lamprey VLRA and VLRB monoclonal antibodies

The long view: a bright past, a brighter future? Forty years of chicken immunology pre- and post-genome

Contact Info

Product

Resources

About