Characterization of Bovine Homologues of Granulysin and NK-lysin

Abstract

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+ γδ 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.

NK-lysin of channel catfish: Gene triplication, sequence variation, and expression analysis

“…Compared with the classical antimicrobial peptides, NK-lysin and granulysin are much larger with 78 and 74 amino acid residues and a globular three-dimensional structure. Since the initial identification of NK-lysin in porcine and granulysin in humans a few years ago, a bovine homolog was recently identified (Endsley et al, 2004).…”

Characterization of a NK-lysin antimicrobial peptide gene from channel catfish

“…Although a bovine homolog has recently been identified [13], the status of the NK-lysin/granulysin antimicrobial class in lower vertebrates is unknown. As part of a larger effort to characterize the catfish innate immune system, we report here identification, molecular cloning and characterization, and expression of a NK-lysin gene from channel catfish (Ictalurus punctatus).…”

Immunity and Vaccination against Tuberculosis in Cattle

“…The circulatory population then rapidly expands and shows an activated profile with increased CD25 + expression. The cells have been shown to be cytotoxic [34] and express perforin [35] and granulysin [22]. Contact dependant interactions between DC and WC1 + γδ cells have demonstrated that DC induce activation of WC1 + γδ cells which leads to IFN-γ release [36].…”

“…The primary cell type performing this function is the CD8 + T cell. Also, other cells which have this cytolytic capacity include NK cells, CD4 + T cells, and γδ T cells [22,29].…”

“…M. bovis-infected DC release chemokines to preferentially recruit the NKp46 + CD2 -population of NK cells with subsequent activation of these cells. NK cells are also cytotoxic through the release of perforin and granulysin leading to killing of M. bovis-infected APCs [21][22][23]. Release of perforin is attained both in association with and without DC involvement [24].…”