T cells in cryptopatch aggregates share TCR γ variable region junctional sequences with γδ cells in the small intestinal epithelium of mice

Bradley S. Podd, Joseph Thoits, Nicholas Whitley, Hao Yuan Cheng, Kimberly L. Kudla, Hiroko Taniguchi, Joanna Halkias, Kerstin Goth, Victoria Camerini

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

The role of cryptopatch aggregates in the development of intestinal intraepithelial lymphocytes (IEL) is a matter of controversy. Therefore, an important question is whether T cells in cryptopatch aggregates are lineally related to IEL. We hypothesized that if γδ+ IEL derive from T cells in cryptopatch aggregates, then a clonal relationship would exist between the two populations. To test this hypothesis, we compared the sequence of rearranged TCR gamma variable region 5 genes in γδ+ IEL and cryptopatch cells. We purified IEL by FACS and cryptopatch cells were isolated from frozen sections of the intestine by laser-assisted microdissection. PCR showed that TCR gamma variable region 5 was rearranged in γδ+ IEL and in CD3+ cryptopatch cells, but not in CD3- cryptopatch cells. DNA sequence analysis showed that the frequency of in-frame junctions in cryptopatch aggregates was at a level consistent with positive selection in both wild-type and athymic nude mice. In addition, the predicted amino acid sequences of V-J junctions present in γδ+ IEL and cryptopatch cells were encoded by identical nucleotide sequences. By contrast, the frequency of in-frame joints was significantly reduced in cryptopatch cells isolated from TCR δ-deficient mice, indicating that the enrichment of in-frame joints in cryptopatch cells must normally depend on expression of surface γδ TCR. Our results are consistent with the hypothesis that a subset of γδ+ IEL are related to T cells in cryptopatch aggregates. The precise role of cryptopatch aggregates in intestinal γδ+ T cell homeostasis still needs to be determined.

Original languageEnglish (US)
Pages (from-to)6532-6542
Number of pages11
JournalJournal of Immunology
Volume176
Issue number11
DOIs
StatePublished - Jun 1 2006
Externally publishedYes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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