Characterization of Bovine Homologues of Granulysin and NK-lysin

• Published in: The Journal of immunology (1950) 2004-08, Vol.173 (4), p.2607-2614
• Main Authors: Endsley, Janice J, Furrer, Jason L, Endsley, Mark A, McIntosh, Mark A, Maue, Alexander C, Waters, W. Ray, Lee, David R, Estes, D. Mark
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antigens, Differentiation, T-Lymphocyte - genetics; Antigens, Differentiation, T-Lymphocyte - immunology; Bacillus; Bacterial Infections - immunology; Base Sequence; Cattle; CD3 Complex - immunology; CD4 Antigens - immunology; CD8 Antigens - immunology; Escherichia coli; Flow Cytometry; Gene Expression - immunology; Humans; Immunoblotting; Leukocytes, Mononuclear - immunology; Mice; Molecular Sequence Data; Mycobacterium bovis; Proteolipids - genetics; Proteolipids - immunology; Reverse Transcriptase Polymerase Chain Reaction; Salmonella enteritidis; Sequence Homology, Amino Acid; Staphylococcus aureus; T-Lymphocytes - immunology
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.173.4.2607

Granulysin and NK-lysin are antimicrobial proteins found in the granules of human and swine cytotoxic lymphocytes. A murine counterpart to granulysin has not been identified to date, indicating the importance of additional models to fully characterize the role of granulysin-like molecules in the immune response to infectious disease. Two partial nucleotide sequences corresponding to the complete functional domain of granulysin and NK-lysin were amplified from bovine PBMC mRNA. Following stimulation with phorbol ester and calcium ionophore, expression of the bovine gene was detected in CD3(+) T cells, CD4(+) T cells, CD8(+) T cells, WC1(+) gammadelta T cells, and PBMC depleted of CD3(+) T cells, but was absent in CD21(+) cells and CD14(+) cells. Intracellular flow cytometry and immunoblotting confirmed the presence of protein corresponding to the bovine granulysin homologue in activated T lymphocytes and PBMC. Synthetic human, bovine, and swine peptides corresponding to the C terminus of helix 2 through helix 3 region of granulysin displayed potent antimicrobial activity against Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, and Mycobacterium bovis bacillus Calmette-Guérin. Human and bovine peptides corresponding to helix 2 displayed antimycobacterial activity against M. bovis bacillus Calmette-Guérin. Expression of the bovine gene was detected in laser microscopy-dissected lymph node lesions from an M. bovis-infected animal. The identification of a biologically active bovine homologue to granulysin demonstrates the potential of the bovine model in characterizing the role of granulysin in the immune response to a variety of infectious agents.