Allelic exclusion frequency analysis and molecular characteristics of immunoglobulins secreted by the hybridomas expressing both allelic genes

Cherepakhin, V. V.; Jackubov, L.Z.; Ibraghimov, Alexander; Rokhlin, O.V.

doi:10.1016/0165-2478(87)90073-3

Cited by 3 publications

(2 citation statements)

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“…This was not observed, and the discrepancy could be explained by differences in the criteria used to score a cell as DE by intracytoplasmic staining. Our findings are consistent with the notion that IgH allelic exclusion is an extremely tightly regulated mechanism, even when compared with that of other Ag receptor genes known to be stringently phenotypically excluded, such as the TCR ␤ (19,20) and ␦ (21) chains and the Ig (2,(22)(23)(24) and (4) L chains. The finding that phenotypic allelically included cells do exist but are kept at very low frequencies raises some questions concerning the selective pressures that shaped allelic exclusion.…”

Section: Discussionsupporting

confidence: 90%

Frequency and Characterization of Phenotypic Ig Heavy Chain Allelically Included IgM-Expressing B Cells in Mice

Barreto

Cumano

2000

The Journal of Immunology

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Ig H chain (IgH) allelic exclusion remains a puzzling topic. Here, we address the following question: Do phenotypic IgH allelically included cells exist in normal mice and, if so, at what frequency? Sorted cells from heterozygous mice were evaluated for the expression of both IgM allotypes by double intracytoplasmic stainings. Dual expressors were found at a frequency of 1 in 104 splenic B cells. These data were confirmed by direct sequencing of IgH-rearranged alleles obtained after single cell (or clone) PCR on dual expressors. Typically, these cells have one rearranged J558 VH whereas, in the other allele, a D-proximal VH gene is used. Interestingly, dual expressors have rearranged IgH alleles with similar CDR3 lengths. These results show that, in contrast to the κ L chain and the TCR β-chain, IgH allelic exclusion is the result of an extremely stringent mechanism. We discuss two non-mutually exclusive scenarios for the origin of IgH dual expressors: 1) IgH allelically included cells arise when the first allele to rearrange productively is unable to form a pre-BCR; dual expressors could be a subset of this population in which, upon conventional L chain rearrangement, both IgH are expressed at the surface; and 2) synchronous rearrangement of the IgH alleles.

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Section: Discussionsupporting

confidence: 90%

Frequency and Characterization of Phenotypic Ig Heavy Chain Allelically Included IgM-Expressing B Cells in Mice

Barreto

Cumano

2000

The Journal of Immunology

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“…In support of the first possibility, the innocuous LC chain is thought to compete with the autoreactive LC for binding of free HCs and thus suppress the assembly and surface transport of the autoreactive BCR, thereby diminishing the recognition of self‐antigens (29, 195). In support of the latter, however, IgL allelic inclusion clearly increases the risk that B cells will secrete autoreactive antibodies, even if the cells are activated by the binding of antigens to the non‐autoreactive BCR specificity.…”

Section: Monospecificity Of B Cells and Immune Recognitionmentioning

confidence: 99%

Allelic exclusion of immunoglobulin genes: models and mechanisms

Vettermann

Schlissel

2010

Immunological Reviews

142

131

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SummaryThe allelic exclusion of immunoglobulin (Ig) genes is one of the most evolutionarily conserved features of the adaptive immune system and underlies the monospecificity of B cells. While much has been learned about how Ig allelic exclusion is established during B-cell development, the relevance of monospecificity to B-cell function remains enigmatic. Here, we review the theoretical models that have been proposed to explain the establishment of Ig allelic exclusion and focus on the molecular mechanisms utilized by developing B cells to ensure the monoallelic expression of Igκ and Igλ light chain genes. We also discuss the physiological consequences of Ig allelic exclusion and speculate on the importance of monospecificity of B cells for immune recognition. Monospecificity of B lymphocytes and Ig allelic exclusionSince Burnet's clonal selection theory of the adaptive immune system, the monospecificity of B lymphocytes has been a central paradigm in explaining the pathogen-specific production of antibodies (1). This paradigm, also known as the 'one B cell -one antibody' rule, is supported by a great body of experimental evidence (2-6). According to Burnet's theory, antibodies displayed as B-cell antigen receptors (BCRs) at the surface of a single B cell contain only one particular antigen-binding site, allowing for the clonal selection of antibody-producing cells by their respective pathogen-associated antigens. Ideally, the monospecific expression of BCRs by B cells and the highly specific BCR/antigen interaction result in an antibody response that targets the pathogen, while avoiding wasted resources and collateral damage.

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Characterization of a Unique Anti-DNA Hybridoma

Blatt

Bill

Glick³

1998

Hybridoma

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We have previously described the isolation and in vitro binding properties of eight anti-DNA monoclonal antibodies (MAbs) from an MRL-lpr mouse. In light of recent reports that have indicated it is possible to isolate multiple MAbs from a single hybridoma, our pathogenic hybridoma, 11F8, was examined for evidence of similar phenomena. Chromosome counting suggested that 11F8 cells are unusual and might indeed be expressing multiple heavy and/or light chains. PCR, cloning, and sequencing of immunoglobulin heavy and light chains indicate that 11F8 displays expression of both gamma 2a and gamma 3 heavy chains at the DNA level. Flow cytometry and amino acid sequencing reveals that expression of multiple isotypes also occurs at the protein level but only a single heavy- and light-chain sequence is able to bind DNA. Based on these results, we conclude that 11F8 is an unusual hybridoma that secretes two distinct heavy and at least one light chain from a single cell, and may represent a trioma, a stable three-cell fusion.

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Allelic exclusion frequency analysis and molecular characteristics of immunoglobulins secreted by the hybridomas expressing both allelic genes

Cited by 3 publications

References 10 publications

Frequency and Characterization of Phenotypic Ig Heavy Chain Allelically Included IgM-Expressing B Cells in Mice

Frequency and Characterization of Phenotypic Ig Heavy Chain Allelically Included IgM-Expressing B Cells in Mice

Allelic exclusion of immunoglobulin genes: models and mechanisms

Characterization of a Unique Anti-DNA Hybridoma

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