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

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

Author(s)
Buonocore, Francesco
Randelli, Elisa
Casani, Daniela
Costantini, Susan
Facchiano, Angelo M.
more
Date Issued
2007
Type
Article
DOI
10.1016/j.fsi.2007.03.013
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 b 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 50-UTR and a 478 bp 30-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 b 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-1b (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.
Additional information
L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/
Citation
Buonocore, F. et al. 2007. Molecular cloning, differential expression and 3D structural analysis of the MHC class-II β chain from sea bass (Dicentrarchus labrax L.). "Fish and Shellfish Immunology" 23(4) 853-866
Subjects

Fish Immmunology

Molecular Biology

Sea bass

Dicentrarchus labrax

Cloning

Polymorphism

Expression analysis

Quantitative PCR

3D structure

Handle
http://hdl.handle.net/2067/1357
File(s)
Thumbnail Image
Name

MHCseabassfin.pdf

Size

735.73 KB

Format

Adobe PDF

Checksum (MD5)

b4637d328d9cee26ee7cc225582c0c58

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Sponsor(s)
This work was supported by the European Commission within the project IMAQUANIM (EC contract number
FOOD-CT-2005-007103).

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