title: Platelet antistaphylococcal responses occur through P2X1 and P2Y12 receptor-induced activation and kinocidin release.
abstract: Platelets (PLTs) act in antimicrobial host defense by releasing PLT microbicidal proteins (PMPs) or PLT kinocidins (PKs). Receptors mediating staphylocidal efficacy and PMP or PK release versus isogenic PMP-susceptible (ISP479C) and -resistant (ISP479R) Staphylococcus aureus strains were examined in vitro. Isolated PLTs were incubated with ISP479C or ISP479R (PLT/S. aureus ratio range, 1:1 to 10,000:1) in the presence or absence of a panel of PLT inhibitors, including P2X and P2Y receptor antagonists of increasingly narrow specificity, and PLT adhesion receptors (CD41, CD42b, and CD62P). PLT-to-S. aureus exposure ratios of > or = 10:1 yielded significant reductions in the viability of both strains. Results from reversed-phase high-performance liquid chromatography indicated that staphylocidal PLT releasates contained PMPs and PKs. At ratios below 10:1, the PLT antistaphylococcal efficacy relative to the intrinsic S. aureus PMP-susceptible or -resistant phenotype diminished. Apyrase (an agent of ADP degradation), suramin (a general P2 receptor antagonist), pyridoxal 5'-phosphonucleotide derivative (a specific P2X(1) antagonist), and cangrelor (a specific P2Y(12) antagonist) mitigated the PLT staphylocidal response against both strains, correlating with reduced levels of PMP and PK release. Specific inhibition occurred in the presence and absence of homologous plasma. The antagonism of the thromboxane A(2), cyclooxygenase-1/cyclooxygenase-2, or phospholipase C pathway or the hindrance of surface adhesion receptors failed to impede PLT anti-S. aureus responses. These results suggest a multifactorial PLT anti-S. aureus response mechanism involving (i) a PLT-to-S. aureus ratio sufficient for activation; (ii) the ensuing degranulation of PMPs, PKs, ADP, and/or ATP; (iii) the activation of P2X(1)/P2Y(12) receptors on adjacent PLTs; and (iv) the recursive amplification of PMP and PK release from these PLTs.
authors.affiliation: Los Angeles Biomedical Research Institute, Harbor-UCLA Medical Center, Torrance, CA 90509, USA.
authors.forename: Darin A
authors.initials: DA
authors.lastname: Trier
authors.forename: Kimberly D
authors.initials: KD
authors.lastname: Gank
authors.forename: Deborah
authors.initials: D
authors.lastname: Kupferwasser
authors.forename: Nannette Y
authors.initials: NY
authors.lastname: Yount
authors.forename: William J
authors.initials: WJ
authors.lastname: French
authors.forename: Alan D
authors.initials: AD
authors.lastname: Michelson
authors.forename: Leon I
authors.initials: LI
authors.lastname: Kupferwasser
authors.forename: Yan Q
authors.initials: YQ
authors.lastname: Xiong
authors.forename: Arnold S
authors.initials: AS
authors.lastname: Bayer
authors.forename: Michael R
authors.initials: MR
authors.lastname: Yeaman
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000276:immunology* / Q000378:metabolism; D018925:Chemokines / Q000096:biosynthesis* / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D015539:Platelet Activation / Q000276:immunology*; D011817:Rabbits; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058469:Receptors, Purinergic P2X; D058925:Receptors, Purinergic P2Y12; D013203:Staphylococcal Infections / Q000276:immunology*; D013211:Staphylococcus aureus / Q000276:immunology
chemical_list: D018925:Chemokines; D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D058925:Receptors, Purinergic P2Y12