Detection of spliced and unspliced forms of germline TCR-V beta transcripts in extrathymic lymphoid sites

Janice L. Abbey, Helen C. O'Neill

Research output: Contribution to journalArticleResearchpeer-review

3 Citations (Scopus)

Abstract

Germline TCR-V beta transcription is commonly considered an event coupled with rearrangement of TCR genes in T cells. The extent of germline V beta transcription is studied here in a range of cell types and in several mouse strains. A sensitive semi-quantitative RT-PCR method was developed to specifically detect germline and not rearranged transcripts. Germline transcription of a range of different V beta genes was detected along with rearranged transcripts in bone marrow, thymus, mesenteric lymph node and spleen. Some transcripts were also detected in low level in non-lymphoid tissues including heart, liver and brain. Expression was also studied in the C57BL/6J-beta(2)microglobulin(-/-) (C57BL/6J-beta M-2(-/-)) mouse model that lacks NK1.1 T cells and predominantly utilises V beta 8.2 in the formation of a TCR. beta M-2(-/-) mice, which lack both CD-dependent NK1.1 T cells and CD8(+) T cells, showed germline TCR-V beta 8 transcription in most tissues indicating that germline transcription is not specifically related to CD1-dependent NK1.1 T cells. In many tissues, multiple transcripts were amplified representing both spliced and unspliced forms of germline V beta. For most V beta genes, the expression of spliced and unspliced forms was equivalent. Given an abundance of unspliced transcripts, the presence of alternative ORFs encoding a novel protein was investigated within the TCR-V beta genes. Sequence analysis of ORFs showed only genes with a high level of similarity to TCR-beta. All data reflect the prevalence of germline transcripts in vivo and raise questions about their functional role. (c) 2007 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)1099-1111
Number of pages13
JournalMolecular Immunology
Volume45
Issue number4
DOIs
Publication statusPublished - Feb 2008
Externally publishedYes

Cite this

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title = "Detection of spliced and unspliced forms of germline TCR-V beta transcripts in extrathymic lymphoid sites",
abstract = "Germline TCR-V beta transcription is commonly considered an event coupled with rearrangement of TCR genes in T cells. The extent of germline V beta transcription is studied here in a range of cell types and in several mouse strains. A sensitive semi-quantitative RT-PCR method was developed to specifically detect germline and not rearranged transcripts. Germline transcription of a range of different V beta genes was detected along with rearranged transcripts in bone marrow, thymus, mesenteric lymph node and spleen. Some transcripts were also detected in low level in non-lymphoid tissues including heart, liver and brain. Expression was also studied in the C57BL/6J-beta(2)microglobulin(-/-) (C57BL/6J-beta M-2(-/-)) mouse model that lacks NK1.1 T cells and predominantly utilises V beta 8.2 in the formation of a TCR. beta M-2(-/-) mice, which lack both CD-dependent NK1.1 T cells and CD8(+) T cells, showed germline TCR-V beta 8 transcription in most tissues indicating that germline transcription is not specifically related to CD1-dependent NK1.1 T cells. In many tissues, multiple transcripts were amplified representing both spliced and unspliced forms of germline V beta. For most V beta genes, the expression of spliced and unspliced forms was equivalent. Given an abundance of unspliced transcripts, the presence of alternative ORFs encoding a novel protein was investigated within the TCR-V beta genes. Sequence analysis of ORFs showed only genes with a high level of similarity to TCR-beta. All data reflect the prevalence of germline transcripts in vivo and raise questions about their functional role. (c) 2007 Elsevier Ltd. All rights reserved.",
author = "Abbey, {Janice L.} and O'Neill, {Helen C.}",
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Detection of spliced and unspliced forms of germline TCR-V beta transcripts in extrathymic lymphoid sites. / Abbey, Janice L.; O'Neill, Helen C.

In: Molecular Immunology, Vol. 45, No. 4, 02.2008, p. 1099-1111.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Detection of spliced and unspliced forms of germline TCR-V beta transcripts in extrathymic lymphoid sites

AU - Abbey, Janice L.

AU - O'Neill, Helen C.

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N2 - Germline TCR-V beta transcription is commonly considered an event coupled with rearrangement of TCR genes in T cells. The extent of germline V beta transcription is studied here in a range of cell types and in several mouse strains. A sensitive semi-quantitative RT-PCR method was developed to specifically detect germline and not rearranged transcripts. Germline transcription of a range of different V beta genes was detected along with rearranged transcripts in bone marrow, thymus, mesenteric lymph node and spleen. Some transcripts were also detected in low level in non-lymphoid tissues including heart, liver and brain. Expression was also studied in the C57BL/6J-beta(2)microglobulin(-/-) (C57BL/6J-beta M-2(-/-)) mouse model that lacks NK1.1 T cells and predominantly utilises V beta 8.2 in the formation of a TCR. beta M-2(-/-) mice, which lack both CD-dependent NK1.1 T cells and CD8(+) T cells, showed germline TCR-V beta 8 transcription in most tissues indicating that germline transcription is not specifically related to CD1-dependent NK1.1 T cells. In many tissues, multiple transcripts were amplified representing both spliced and unspliced forms of germline V beta. For most V beta genes, the expression of spliced and unspliced forms was equivalent. Given an abundance of unspliced transcripts, the presence of alternative ORFs encoding a novel protein was investigated within the TCR-V beta genes. Sequence analysis of ORFs showed only genes with a high level of similarity to TCR-beta. All data reflect the prevalence of germline transcripts in vivo and raise questions about their functional role. (c) 2007 Elsevier Ltd. All rights reserved.

AB - Germline TCR-V beta transcription is commonly considered an event coupled with rearrangement of TCR genes in T cells. The extent of germline V beta transcription is studied here in a range of cell types and in several mouse strains. A sensitive semi-quantitative RT-PCR method was developed to specifically detect germline and not rearranged transcripts. Germline transcription of a range of different V beta genes was detected along with rearranged transcripts in bone marrow, thymus, mesenteric lymph node and spleen. Some transcripts were also detected in low level in non-lymphoid tissues including heart, liver and brain. Expression was also studied in the C57BL/6J-beta(2)microglobulin(-/-) (C57BL/6J-beta M-2(-/-)) mouse model that lacks NK1.1 T cells and predominantly utilises V beta 8.2 in the formation of a TCR. beta M-2(-/-) mice, which lack both CD-dependent NK1.1 T cells and CD8(+) T cells, showed germline TCR-V beta 8 transcription in most tissues indicating that germline transcription is not specifically related to CD1-dependent NK1.1 T cells. In many tissues, multiple transcripts were amplified representing both spliced and unspliced forms of germline V beta. For most V beta genes, the expression of spliced and unspliced forms was equivalent. Given an abundance of unspliced transcripts, the presence of alternative ORFs encoding a novel protein was investigated within the TCR-V beta genes. Sequence analysis of ORFs showed only genes with a high level of similarity to TCR-beta. All data reflect the prevalence of germline transcripts in vivo and raise questions about their functional role. (c) 2007 Elsevier Ltd. All rights reserved.

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JF - Immunochemistry

SN - 0161-5890

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