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Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells

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Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells. / Winter, Manuel; Kashani, Elham; Chennupati, Vijaykumar; Föhse, Lisa; Prinz, Immo.

In: IMMUNOL CELL BIOL, Vol. 92, No. 5, 06.2014, p. 409-16.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Winter, M, Kashani, E, Chennupati, V, Föhse, L & Prinz, I 2014, 'Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells', IMMUNOL CELL BIOL, vol. 92, no. 5, pp. 409-16. https://doi.org/10.1038/icb.2013.105

APA

Winter, M., Kashani, E., Chennupati, V., Föhse, L., & Prinz, I. (2014). Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells. IMMUNOL CELL BIOL, 92(5), 409-16. https://doi.org/10.1038/icb.2013.105

Vancouver

Winter M, Kashani E, Chennupati V, Föhse L, Prinz I. Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells. IMMUNOL CELL BIOL. 2014 Jun;92(5):409-16. https://doi.org/10.1038/icb.2013.105

Bibtex

@article{667957987f6140cd99d733d336aa4afe,
title = "Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells",
abstract = "T-cell receptor α (TCRα) chain rearrangement is not constrained by allelic exclusion and thus αβ T cells frequently have rearranged both alleles of this locus. Thereby, stepwise secondary rearrangements of both TCRα loci further increase the odds for generation of an α-chain that can be positively selected in combination with a pre-existing TCRβ chain. Previous studies estimated that approximately 2-12% of murine and human αβ T cells still carry one TCRα locus in germline configuration, which must comprise a partially or even fully rearranged TCRδ locus. However, these estimates are based on a relatively small amount of individual αβ T-cell clones and αβ T-cell hybridomas analyzed to date. To address this issue more accurately, we made use of a mouse model, in which a fluorescent reporter protein is introduced into the constant region of the TCRδ locus. In this TcrdH2BeGFP system, fluorescence emanating from retained TCRδ loci enabled us to quantify monoallelically rearranged αβ T cells on a single-cell basis. Via fluorescence-activated cell sorting analysis, we determined the frequency of monoallelic TCRα rearrangements to be 1.7% in both peripheral CD4(+) and CD8(+) αβ T cells. Furthermore, we found a skewed 5' Jα gene utilization of the rearranged TCRα allele in T cells with monoallelic TCRα rearrangements. This is in line with previous descriptions of a tight interallelic positional coincidence of Jα gene segments used on both TCRα alleles. Finally, analysis of T cells from transgenic mice harboring only one functional TCRα locus implied the existence of very rare unusual translocation or episomal reintegration events of formerly excised TCRδ loci. ",
keywords = "Alleles, Animals, Gene Expression, Gene Frequency, Gene Rearrangement, T-Lymphocyte, Genes, Reporter, Genetic Loci, Mice, Mice, Transgenic, Receptors, Antigen, T-Cell, alpha-beta/genetics, T-Lymphocyte Subsets/metabolism",
author = "Manuel Winter and Elham Kashani and Vijaykumar Chennupati and Lisa F{\"o}hse and Immo Prinz",
year = "2014",
month = jun,
doi = "10.1038/icb.2013.105",
language = "English",
volume = "92",
pages = "409--16",
journal = "IMMUNOL CELL BIOL",
issn = "0818-9641",
publisher = "John Wiley & Sons",
number = "5",

}

RIS

TY - JOUR

T1 - Visualization and quantification of monoallelic TCRα gene rearrangement in αβ T cells

AU - Winter, Manuel

AU - Kashani, Elham

AU - Chennupati, Vijaykumar

AU - Föhse, Lisa

AU - Prinz, Immo

PY - 2014/6

Y1 - 2014/6

N2 - T-cell receptor α (TCRα) chain rearrangement is not constrained by allelic exclusion and thus αβ T cells frequently have rearranged both alleles of this locus. Thereby, stepwise secondary rearrangements of both TCRα loci further increase the odds for generation of an α-chain that can be positively selected in combination with a pre-existing TCRβ chain. Previous studies estimated that approximately 2-12% of murine and human αβ T cells still carry one TCRα locus in germline configuration, which must comprise a partially or even fully rearranged TCRδ locus. However, these estimates are based on a relatively small amount of individual αβ T-cell clones and αβ T-cell hybridomas analyzed to date. To address this issue more accurately, we made use of a mouse model, in which a fluorescent reporter protein is introduced into the constant region of the TCRδ locus. In this TcrdH2BeGFP system, fluorescence emanating from retained TCRδ loci enabled us to quantify monoallelically rearranged αβ T cells on a single-cell basis. Via fluorescence-activated cell sorting analysis, we determined the frequency of monoallelic TCRα rearrangements to be 1.7% in both peripheral CD4(+) and CD8(+) αβ T cells. Furthermore, we found a skewed 5' Jα gene utilization of the rearranged TCRα allele in T cells with monoallelic TCRα rearrangements. This is in line with previous descriptions of a tight interallelic positional coincidence of Jα gene segments used on both TCRα alleles. Finally, analysis of T cells from transgenic mice harboring only one functional TCRα locus implied the existence of very rare unusual translocation or episomal reintegration events of formerly excised TCRδ loci.

AB - T-cell receptor α (TCRα) chain rearrangement is not constrained by allelic exclusion and thus αβ T cells frequently have rearranged both alleles of this locus. Thereby, stepwise secondary rearrangements of both TCRα loci further increase the odds for generation of an α-chain that can be positively selected in combination with a pre-existing TCRβ chain. Previous studies estimated that approximately 2-12% of murine and human αβ T cells still carry one TCRα locus in germline configuration, which must comprise a partially or even fully rearranged TCRδ locus. However, these estimates are based on a relatively small amount of individual αβ T-cell clones and αβ T-cell hybridomas analyzed to date. To address this issue more accurately, we made use of a mouse model, in which a fluorescent reporter protein is introduced into the constant region of the TCRδ locus. In this TcrdH2BeGFP system, fluorescence emanating from retained TCRδ loci enabled us to quantify monoallelically rearranged αβ T cells on a single-cell basis. Via fluorescence-activated cell sorting analysis, we determined the frequency of monoallelic TCRα rearrangements to be 1.7% in both peripheral CD4(+) and CD8(+) αβ T cells. Furthermore, we found a skewed 5' Jα gene utilization of the rearranged TCRα allele in T cells with monoallelic TCRα rearrangements. This is in line with previous descriptions of a tight interallelic positional coincidence of Jα gene segments used on both TCRα alleles. Finally, analysis of T cells from transgenic mice harboring only one functional TCRα locus implied the existence of very rare unusual translocation or episomal reintegration events of formerly excised TCRδ loci.

KW - Alleles

KW - Animals

KW - Gene Expression

KW - Gene Frequency

KW - Gene Rearrangement, T-Lymphocyte

KW - Genes, Reporter

KW - Genetic Loci

KW - Mice

KW - Mice, Transgenic

KW - Receptors, Antigen, T-Cell, alpha-beta/genetics

KW - T-Lymphocyte Subsets/metabolism

U2 - 10.1038/icb.2013.105

DO - 10.1038/icb.2013.105

M3 - SCORING: Journal article

C2 - 24418818

VL - 92

SP - 409

EP - 416

JO - IMMUNOL CELL BIOL

JF - IMMUNOL CELL BIOL

SN - 0818-9641

IS - 5

ER -