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Title: Expressed gene sequence of the IFN-gamma-response chemokine CXCL9 of cattle, horses, and swine

Author
item HUDGENS, EDWARD - University Of Massachusetts
item TOMPKINS, DANNIELLE - University Of Massachusetts
item Boyd, Patricia
item Lunney, Joan
item HOROHOV, DAVID - University Of Kentucky
item BALDWIN, CYNTHIA - University Of Kentucky

Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/15/2011
Publication Date: 6/15/2011
Citation: Hudgens, E., Tompkins, D., Boyd, P., Lunney, J.K., Horohov, D., Baldwin, C. 2011. Expressed gene sequence of the IFN-gamma-response chemokine CXCL9 of cattle, horses, and swine. Veterinary Immunology and Immunopathology. 141(3-4):317-321.

Interpretive Summary: Studies of basic immunity and vaccine and disease responses in swine, horses and cattle require basic tools to quantitate cellular trafficking and immune responses. There are limited commercial reagents to measure immune chemoattractant molecules. The U.S. Veterinary Immune Reagent Network(US VIRN www.vetimm.org) was established to address this gap. This report describes the cloning and characterization of expressed gene sequences of a chemokine, CXCL9, from bovine, equine, and swine. Using RNA obtained from peripheral blood mononuclear cell (PBMC) or other tissues the cDNA was prepared and cloned and sequences compared to other species. The bovine coding region was 378 nucleotides in length, while the equine and swine coding regions were 381 nucleotides. Mapping showed that all three sequences were coded for in 4 exons in the genome as are the human and mouse genes. The bovine, equine, and swine coding regions shared 83%, 86%, and 84% homology with human CXCL9, respectively, and all three were 74% homologous with mouse CXCL9 while comparison with other human and mouse C-X-C motif ligand sequences showed less than 50% homology. Using computer models a comparision of the nucleotide sequences of CXCL9 (cladogram) was generated and showed that the bovine, equine and swine sequences were most closely related to one another and that the human sequence clustered more closely with them than did any to the mouse sequence. These relationships were preserved when the deduced amino acid sequences were evaluated and all sequences showed conservation of the characteristic 4 cysteines. This work sets the stage for further work with these molecules. An integral goal of the U.S. Veterinary Immune Reagent Network is to develop reagents for investigating diseases in livestock species, poultry, and fish.

Technical Abstract: This report describes the cloning and characterization of expressed gene sequences of bovine, equine, and swine CXCL9 from RNA obtained from peripheral blood mononuclear cell (PBMC) or other tissues. The bovine coding region was 378 nucleotides in length, while the equine and swine coding regions were 381 nucleotides. Mapping showed that all three sequences were coded for in 4 exons in the genome as are the human and mouse genes. The bovine, equine, and swine coding regions shared 83%, 86%, and 84% homology with human CXCL9, respectively, and all three were 74% homologous with mouse CXCL9 while comparison with other human and mouse C-X-C motif ligand sequences showed less than 50% homology. Cladogram comparision of the nucleotide sequences of CXCL9 showed that the bovine, equine and swine sequences were most closely related to one another and that the human sequence clustered more closely with them than did any to the mouse sequence. These relationships were preserved when the deduced amino acid sequences were evaluated and all sequences showed conservation of the characteristic 4 cysteines. This work sets the stage for further work with these molecules. An integral goal of the U.S. Veterinary Immune Reagent Network is to develop reagents for investigating diseases in livestock species, poultry, and fish.