Molecular cloning, differential expression and 3D structural analysis of the MHC class-II β chain from sea bass (Dicentrarchus labrax L.)

Francesco Buonocore, Elisa Randelli, Daniela Casani, Susan Costantini, Angelo Facchiano, Giuseppe Scapigliati, Renè J M Stet

Research output: Contribution to journalArticle

Abstract

The major histocompatibility complex class I and II molecules (MHC-I and MHC-II) play a pivotal role in vertebrate immune response to antigenic peptides. In this paper we report the cloning and sequencing of the MHC class II β chain from sea bass (Dicentrarchus labrax L.). The six obtained cDNA sequences (designated as Dila-DAB) code for 250 amino acids, with a predicted 21 amino acid signal peptide and contain a 28 bp 5′-UTR and a 478 bp 3′-UTR. A multiple alignment of the predicted translation of the Dila-DAB sequences was assembled together with other fish and mammalian sequences and it showed the conservation of most amino acid residues characteristic of the MHC class II β chain structure. The highest basal Dila-DAB expression was found in gills, followed by gut and thymus, lower mRNA levels were found in spleen, peripheral blood leucocytes (PBL) and liver. Stimulation of head kidney leukocytes with LPS for 4 h showed very little difference in the Dila-DAB expression, but after 24 h the Dila-DAB level decreased to a large extent and the difference was statistically significant. Stimulation of head kidney leukocytes with different concentrations of rIL-1β (ranging from 0 to 100 ng/ml) resulted in a dose-dependent reduction of the Dila-DAB expression. Moreover, two 3D Dila-DAB*0101 homology models were obtained based on crystallographic mouse MHC-II structures complexed with D10 T-cell antigen receptor or human CD4; features and differences between the models were evaluated and discussed. Taken together these results are of interest as MHC-II structure and function, molecular polymorphism and differential gene expression are in correlation with disease resistance to virus and bacteria in teleost fish.

Original languageEnglish
Pages (from-to)853-866
Number of pages14
JournalFish and Shellfish Immunology
Volume23
Issue number4
DOIs
Publication statusPublished - Oct 2007

Keywords

  • 3D structure
  • chain
  • Cloning
  • Dicentrarchus labrax
  • Expression analysis
  • Major histocompatibility complex class (MHC) II β
  • Polymorphism
  • Quantitative PCR
  • Sea bass

ASJC Scopus subject areas

  • Aquatic Science
  • Immunology

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