10.6084/m9.figshare.5121280.v1 Kobayashi S.D. Kobayashi S.D. Braughton K.R. Braughton K.R. Palazzolo-Ballance A.M. Palazzolo-Ballance A.M. Kennedy A.D. Kennedy A.D. Sampaio E. Sampaio E. Kristosturyan E. Kristosturyan E. Whitney A.R. Whitney A.R. Sturdevant D.E. Sturdevant D.E. Dorward D.W. Dorward D.W. Holland S.M. Holland S.M. Supplementary Material for: Rapid Neutrophil Destruction following Phagocytosis of Staphylococcus aureus Karger Publishers 2010 Neutrophils Bacterial infections Host defense Staphylococcus aureus 2010-06-26 00:00:00 Dataset https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Rapid_Neutrophil_Destruction_following_Phagocytosis_of_Staphylococcus_aureus/5121280 Mechanisms underlying the enhanced virulence phenotype of community-associated methicillin-resistant <i>Staphylococcus aureus</i> (CA-MRSA) are incompletely defined, but presumably include evasion of killing by human polymorphonuclear leukocytes (PMNs or neutrophils). To better understand this phenomenon, we investigated the basis of rapid PMN lysis after phagocytosis of USA300, a prominent CA-MRSA strain. Survival of USA300 clinical isolates after phagocytosis ultimately resulted in neutrophil lysis. PMNs containing ingested USA300 underwent morphological changes consistent with apoptosis, but lysed rapidly thereafter (within 6 h), whereas cells undergoing FAS-mediated apoptosis or phagocytosis-induced cell death remained intact. Phagosome membranes remained intact until the point of PMN destruction, suggesting lysis was not caused by escape of <i>S. aureus</i> from phagosomes or the cytolytic action of pore-forming toxins. Microarray analysis of the PMN transcriptome after phagocytosis of representative community-associated <i>S. aureus</i> and healthcare-associated MRSA strains revealed changes unique to community-associated <i>S. aureus</i> strains, such as upregulation of transcripts involved in regulation of calcium homeostasis. Collectively, the data suggest that neutrophil destruction after phagocytosis of USA300 is in part a form of programmed necrosis rather than direct lysis by <i>S. aureus</i> pore-forming toxins. We propose that the ability of CA-MRSA strains to induce programmed necrosis of neutrophils is a component of enhanced virulence.