Diversity, molecular characterization and expression of T cell receptor γ in a teleost fish, the sea bass (Dicentrarchus labrax, L)

PLoS One. 2012;7(10):e47957. doi: 10.1371/journal.pone.0047957. Epub 2012 Oct 25.

Abstract

Two lineages of T cells, expressing either the αβ T cell receptor (TR) or the γδ TR, exist in Gnathostomes. The latter type of T cells account for 1-10 % of T cells in blood and up to 30 % in the small intestine. They may recognize unconventional antigens (phosphorylated microbial metabolites, lipid antigens) without the need of major histocompatibility class I (MH1) or class II (MH2) presentation. In this work we have described cloning and structural characterization of TR -chain (TRG) from the teleost Dicentrarchus labrax. Further, by means of quantitative PCR analysis, we analyzed TRG expression levels both in poly I:C stimulated leukocytes in vitro, and following infection with betanodavirus in vivo. Two full length cDNAs relative to TRG, with the highest peptide and nucleotide identity with Japanese flounder, were identified. A multiple alignment analysis showed the conservation of peptides fundamental for TRG biological functions, and of the FGXG motif in the FR4 region, typical of most TR and immunoglobulin light chains. A 3D structure consisting of two domains mainly folded as beta strands with a sandwich architecture for each domain was also reported. TRG CDR3 of 8-18 AA in length and diversity in the TRG rearrangements expressed in thymus and intestine for a given V/C combination were evidenced by junction length spectratyping. TRG mRNA expression levels were high in basal conditions both in thymus and intestine, while in kidney and gut leukocytes they were up-regulated after in vitro stimulation by poly I:C. Finally, in juveniles the TRG expression levels were up-regulated in the head kidney and down-regulated in intestine after in vivo infection with betanodavirus. Overall, in this study the involvement of TRG-bearing T cells during viral stimulation was described for the first time, leading to new insights for the identification of T cell subsets in fish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bass
  • DNA Primers / genetics
  • DNA, Complementary / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation*
  • Genes, T-Cell Receptor gamma / genetics*
  • Genetic Variation
  • Leukocytes / metabolism
  • Models, Genetic
  • Models, Molecular
  • Molecular Sequence Data
  • Peptides / chemistry
  • Phosphorylation
  • Receptors, Antigen, T-Cell, gamma-delta / metabolism*
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Thymus Gland / metabolism
  • Tissue Distribution

Substances

  • DNA Primers
  • DNA, Complementary
  • Peptides
  • Receptors, Antigen, T-Cell, gamma-delta

Grants and funding

This work was partially supported by the European Commission within the FP6 project IMAQUANIM (EC contract number FOOD-CT-2005-007103), and the European Community’s Seventh Framework Program (FP7/2007-13) under Grant Agreement 222719 LIFECYCLE. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.