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Title: Expressed gene sequence and bioactivity of the IFN-gamma-response chemokine CXCL11 of swine and cattle

Author
item Boyd, Patricia
item HUDGENS, EDWARD - University Of Massachusetts
item LOFTUS, JOHN - University Of Massachusetts
item TOMPKINS, DANNIELLE - University Of Massachusetts
item WYSOCKI, MICHAL - US Department Of Agriculture (USDA)
item KAKACH, LAURA - Kingfisher Biotech, Inc
item LA BRESH, JOANNA - Kingfisher Biotech, Inc
item BALDWIN, CYNTHIA - University Of Massachusetts
item Lunney, Joan

Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/11/2010
Publication Date: 6/1/2010
Citation: Boyd, P., Hudgens, E., Loftus, J.P., Tompkins, D., Wysocki, M., Kakach, L., La Bresh, J., Baldwin, C.L., Lunney, J.K. 2010. Expressed gene sequence and bioactivity of the IFN-gamma-response chemokine CXCL11 of swine and cattle. Veterinary Immunology and Immunopathology. 136(1-2):170-175.

Interpretive Summary: To investigate basic immunity and vaccine and disease responses in swine and cattle it is essential to have tools to quantitate 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 the swine and bovine interferon-gamma inducible chemokine CXCL11, or I-TAC. This chemokine is associated with T helper 1-type immune responses that help livestock resist viral and bacterial infections. The coding regions of both swine and cattle CXCL11 cDNA sequences were 303 nucleotides in length; each is coded for in four exons in the genome. The bovine coding region shared 82% and 70% homology with human and mouse CXCL11, respectively, and the swine coding region 84% and 72% homology, respectively. Each cDNA was cloned into plasmids and transfected into Pichia (yeast) and the resultant expressed protein purified. Biological activity of each purified chemokine was affirmed by chemotaxis assays. Both swine and bovine CXCL11 were chemotactic for mitogen and IL-2 stimulated peripheral blood mononuclear cells. This is the first report for bioactivity of this chemokine in livestock species and affirmation of bioactivity of the yeast expressed protein products. This work provides valuable new reagents for investigating swine and cattle vaccine and disease responses.

Technical Abstract: This report describes the cloning and characterization of expressed gene sequences of the swine and bovine interferon-gamma inducible chemokine CXCL11, or I-TAC, associated with T helper 1-type immune responses, and affirmation of bioactivity of their yeast expressed protein products. The coding regions of both cDNA sequences were 303 nucleotides in length; each is coded for in four exons in the genome. The bovine coding region shared 82% and 70% homology with human and mouse CXCL11, respectively, and the swine coding region 84% and 72% homology, respectively. As expected the swine and bovine CXCL11 sequences showed less homology with other human and mouse C-X-C motif chemokine sequences. Each cDNA was cloned into plasmids and transfected into Pichia (yeast) and the resultant expressed protein purified. Biological activity of each purified chemokine was affirmed by chemotaxis assays. Both swine and bovine CXCL11 were chemotactic for mitogen and IL-2 stimulated peripheral blood mononuclear cells. This is the first report for bioactivity of this chemokine in livestock species. This work provides valuable new reagents for investigating basic immunity as well as vaccine and disease responses in swine and cattle, goals of the U.S. Veterinary Immune Reagent Network which supported this effort.