Abstract
Infection with Mycobacterium avium subspecies paratuberculosis (MAP) is complex, but little is known about the role that natural killer (NK) cells play. In the present study, four bovine NK-lysin peptides were synthesized to evaluate their bactericidal activity against MAP. The results demonstrated that bNK-lysin peptides were directly bactericidal against MAP, with bNK1 and bNK2A being more potent than bNK2B and bNK2C. Mechanistically, transmission electron microscopy revealed that the incubation of MAP with bNK2A resulted in extensive damage to cell membranes and cytosolic content leakage. Furthermore, the addition of bNK2A linked with a cell-penetrating peptide resulted in increased MAP killing in a macrophage model.
Highlights
Infection with Mycobacterium avium subspecies paratuberculosis (MAP) is complex, but little is known about the role that natural killer (NK) cells play
Studies with any of the four bNK-lysin peptides have not been reported for other mycobacterial pathogens, including MAP
Only live bacterial DNA can be amplified by PMA-quantitative PCR (qPCR)
Summary
Infection with Mycobacterium avium subspecies paratuberculosis (MAP) is complex, but little is known about the role that natural killer (NK) cells play. Granulysin and NK-lysins are found in the granules of human, porcine and bovine cytotoxic T-cells (CTLs) and NK cells [12, 14, 15]. One of the earliest reports of the antimycobacterial activity of granulysin demonstrated that CTLs killed intracellular pathogens such as M. tuberculosis in a granule-dependent manner [12]. This was followed by the finding that active regions of porcine NK-lysin and human granulysin peptides were effective in killing M. tuberculosis, as well as Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli [16]. Mechanisms of innate and adaptive immunity have been well defined for many mycobacterial pathogens, but there is a lack of information regarding the potential bactericidal activity of NK-lysins, for cattle
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