Loratadine Decreases Methicillin-Resistant Staphylococcus aureus Virulence through Global Hemolysin Regulators vraSR and agr

Abstract Methicillin-resistantStaphylococcus aureus(MRSA) is a bacterial pathogen that relies on expression of a wide range of virulence factors. Hemolysins are a diverse collection of cytolytic toxins that promote virulence by being excreted and lysing host blood cells. We previously reported on novel antivirulence compounds that modulate hemolysis in vitro, including a brominated carbazole (compound 8) and the active ingredient in Claritin (loratadine). We analyzed expression and activity of MRSA hemolysins in two hospital-acquired strains. Even with the same staphylococcal cassette chromosome mec (SCCmec) type, the strains displayed unique responses to loratadine where both repression and elevation of hemolysis occurred. These compounds are potentially binding and regulating the master regulatory protein, Stk1, which has known ties to hemolysis inS. aureus. We hypothesized that the modulation of hemolysis by these putative Stk1 inhibitors would extend to community-acquired strains of MRSA. Furthermore, we wanted to determine if Stk1 was the only hemolysis regulatory protein being engaged by these compounds. To test this, we examined compound 8 and loratadine effects on two different USA300 strains, in addition to transposon mutants for a range of hemolysis regulatory genes. We observed strain-specific regulation of hemolysis by Stk1. Loratadine continued to downregulate both alpha and beta hemolytic activity in USA300 JE2 in both Stk1-dependent and independent pathways. Additionally, we report that this antivirulence compound effectively disrupts hemolysis by modulating both agr and vraSR global regulators, contributing to widespread cytotoxin repression. Both alpha hemolysin (hla) and RNAIII/delta hemolysin (hld) mRNA reductions occurred in vitro and in a human cell line model of MRSA infection. Together, these results expand our knowledge of loratadine’s mode of action as a virulence modulator.