Rearrangement and Junctional-Site Sequence Analyses of T-Cell Receptor Gamma Genes in Intestinal Intraepithelial Lymphocytes from Murine Athymic Chimeras

Michael Whetsell, R Lee Mosley, Lynne Whetsell, Frederick V. Schaefer, Kenton S. Miller, John R. Klein

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The molecular organization of rearranged T-cell receptor (TCR) γ genes in intraepithelial lymphocytes (IEL) was studied in athymic radiation chimeras and was compared with the organization of γ gene rearrangements in IEL from thymus-bearing animals by polymerase chain reaction and by sequence analyses of DNA spanning the junction of the variable (V) and joining (J) genes. In both thymus-bearing mice and athymic chimeras, IEL V-J γ-gene rearrangements occurred for Vγ1.2, Vγ2, and Vγ5 but not for Vγ3 or Vγ4. Sequence analyses of cloned V-J polymerase chain reaction-amplified products indicated that in both thymus-bearing mice and athymic chimeras, rearrangement of Vγ1.2 and Vγ5 resulted in in-frame as well as out-of-frame genes, whereas nearly all Vγ2 rearrangements were out of frame from either type of animal. V-segment nucleotide removal occurred in most Vγ1.2, Vγ2, and Vγ5 rearrangements; J-segment nucleotide removal was common in Vγ1.2 but not in Vγ2 or Vγ5 rearrangements. N-segment nucleotide insertions were present in Vγ1.2, Vγ2, and Vγ5 IEL rearrangements in both thymus-bearing mice and athymic chimeras, resulting in a predominant in-frame sequence for Vγ5 and a predominant out-of-frame sequence for Vγ2 genes. These findings demonstrate that (i) TCR γ-gene rearrangement occurs extrathymically in IEL, (ii) rearrangements of TCR γ genes involve the same V genes regardless of thymus influence; and (iii) the thymus does not determine the degree to which functional or nonfunctional rearrangements occur in IEL.

Original languageEnglish (US)
Pages (from-to)5902-5909
Number of pages8
JournalMolecular and cellular biology
Volume11
Issue number12
DOIs
StatePublished - Jan 1 1991

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T-Cell Receptor gamma Genes
Thymus Gland
Sequence Analysis
Lymphocytes
T-Cell Receptor Genes
Nude Mice
Nucleotides
Gene Rearrangement
Genes
Radiation Chimera
T-Lymphocyte Gene Rearrangement
Polymerase Chain Reaction
DNA Sequence Analysis

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Rearrangement and Junctional-Site Sequence Analyses of T-Cell Receptor Gamma Genes in Intestinal Intraepithelial Lymphocytes from Murine Athymic Chimeras. / Whetsell, Michael; Mosley, R Lee; Whetsell, Lynne; Schaefer, Frederick V.; Miller, Kenton S.; Klein, John R.

In: Molecular and cellular biology, Vol. 11, No. 12, 01.01.1991, p. 5902-5909.

Research output: Contribution to journalArticle

Whetsell, Michael ; Mosley, R Lee ; Whetsell, Lynne ; Schaefer, Frederick V. ; Miller, Kenton S. ; Klein, John R. / Rearrangement and Junctional-Site Sequence Analyses of T-Cell Receptor Gamma Genes in Intestinal Intraepithelial Lymphocytes from Murine Athymic Chimeras. In: Molecular and cellular biology. 1991 ; Vol. 11, No. 12. pp. 5902-5909.
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abstract = "The molecular organization of rearranged T-cell receptor (TCR) γ genes in intraepithelial lymphocytes (IEL) was studied in athymic radiation chimeras and was compared with the organization of γ gene rearrangements in IEL from thymus-bearing animals by polymerase chain reaction and by sequence analyses of DNA spanning the junction of the variable (V) and joining (J) genes. In both thymus-bearing mice and athymic chimeras, IEL V-J γ-gene rearrangements occurred for Vγ1.2, Vγ2, and Vγ5 but not for Vγ3 or Vγ4. Sequence analyses of cloned V-J polymerase chain reaction-amplified products indicated that in both thymus-bearing mice and athymic chimeras, rearrangement of Vγ1.2 and Vγ5 resulted in in-frame as well as out-of-frame genes, whereas nearly all Vγ2 rearrangements were out of frame from either type of animal. V-segment nucleotide removal occurred in most Vγ1.2, Vγ2, and Vγ5 rearrangements; J-segment nucleotide removal was common in Vγ1.2 but not in Vγ2 or Vγ5 rearrangements. N-segment nucleotide insertions were present in Vγ1.2, Vγ2, and Vγ5 IEL rearrangements in both thymus-bearing mice and athymic chimeras, resulting in a predominant in-frame sequence for Vγ5 and a predominant out-of-frame sequence for Vγ2 genes. These findings demonstrate that (i) TCR γ-gene rearrangement occurs extrathymically in IEL, (ii) rearrangements of TCR γ genes involve the same V genes regardless of thymus influence; and (iii) the thymus does not determine the degree to which functional or nonfunctional rearrangements occur in IEL.",
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AB - The molecular organization of rearranged T-cell receptor (TCR) γ genes in intraepithelial lymphocytes (IEL) was studied in athymic radiation chimeras and was compared with the organization of γ gene rearrangements in IEL from thymus-bearing animals by polymerase chain reaction and by sequence analyses of DNA spanning the junction of the variable (V) and joining (J) genes. In both thymus-bearing mice and athymic chimeras, IEL V-J γ-gene rearrangements occurred for Vγ1.2, Vγ2, and Vγ5 but not for Vγ3 or Vγ4. Sequence analyses of cloned V-J polymerase chain reaction-amplified products indicated that in both thymus-bearing mice and athymic chimeras, rearrangement of Vγ1.2 and Vγ5 resulted in in-frame as well as out-of-frame genes, whereas nearly all Vγ2 rearrangements were out of frame from either type of animal. V-segment nucleotide removal occurred in most Vγ1.2, Vγ2, and Vγ5 rearrangements; J-segment nucleotide removal was common in Vγ1.2 but not in Vγ2 or Vγ5 rearrangements. N-segment nucleotide insertions were present in Vγ1.2, Vγ2, and Vγ5 IEL rearrangements in both thymus-bearing mice and athymic chimeras, resulting in a predominant in-frame sequence for Vγ5 and a predominant out-of-frame sequence for Vγ2 genes. These findings demonstrate that (i) TCR γ-gene rearrangement occurs extrathymically in IEL, (ii) rearrangements of TCR γ genes involve the same V genes regardless of thymus influence; and (iii) the thymus does not determine the degree to which functional or nonfunctional rearrangements occur in IEL.

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