Activated platelets kill Staphylococcus aureus, but not Streptococcus pneumoniae ‐ The role of FcγRIIa and platelet factor 4/heparin‐antibodies

BACKGROUND: Heparin induced thrombocytopenia (HIT) is likely a misdirected bacterial host defense mechanism. Platelet factor 4 (PF4) binds to polyanions on bacterial surfaces exposing neo-epitopes to which HIT-antibodies bind. Platelets are activated by the resulting immune complexes via FcgammaRIIA, release bactericidal substances and kill Gram-negative Escherichia coli. OBJECTIVES: To assess the role of PF4, anti-PF4/H antibodies and FcgammaRIIa in killing of Gram-positive bacteria by platelets. METHODS: Binding of PF4 to protein-A deficient Staphylococcus aureus (SA113Deltaspa) and non-encapsulated Streptococcus pneumoniae (D39Deltacps) and its conformational change were assessed by flow cytometry using monoclonal (KKO,5B9) and patient derived anti-PF4/H antibodies. Killing of bacteria was quantified by counting colony forming units (cfu) after incubation with platelets or platelet releasate. Using flow cytometry, platelet activation (CD62P-expression, PAC-1 binding) and phosphatidylserine (PS)-exposure were analyzed. RESULTS: Monoclonal and patient-derived anti-PF4/H antibodies bound in the presence of PF4 to both S.aureus and S.pneumoniae (1.6-fold increased fluorescence signal for human anti-PF4/H antibodies to 24.0-fold increase for KKO). S.aureus (5.5x10(4) cfu/ml) was efficiently killed by platelets (2.7x10(4) cfu/ml) or their releasate (2.9x10(4) cfu/ml). Killing was not further enhanced by PF4 or anti-PF4/H antibodies. Blocking FcgammaRIIa had no impact on killing of S.aureus by platelets. In contrast, S.pneumoniae was not killed by platelets or releasate. Instead, after incubation with pneumococci platelets were unresponsive to TRAP-6 stimulation and exposed high levels of PS. CONCLUSIONS: Anti-PF4/H antibodies seem to have only a minor role for direct killing of Gram-positive bacteria by platelets. S.aureus is killed by platelets or platelet releasate. In contrast, S.pneumoniae affect platelet viability.