Identification, cloning and characterization of SpEX exotoxin produced by Staphylococcus pseudintermedius

Abouelkhair, Mohamed A.; Bemis, David A.; Giannone, Richard J.; Frank, Linda A.; Kania, Stephen A.; Bae, Taeok

doi:10.1371/journal.pone.0220301

Identification, cloning and characterization of SpEX exotoxin produced by Staphylococcus pseudintermedius

Journal Article · Mon Jul 29 00:00:00 EDT 2019 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0220301· OSTI ID:1561612

^[1]; Bemis, David A. ^[2]; ^[3]; Frank, Linda A. ^[2]; ^[2]; Bae, Taeok ^[4]

Univ. of Tennessee, Knoxville, TN (United States); Univ. of Sadat City, Menoufia (Egypt)
Univ. of Tennessee, Knoxville, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Indiana Univ., Bloomington, IN (United States)

Staphylococci have evolved numerous strategies to evade their hosts’ immune systems. Some staphylococcal toxins target essential components of host innate immunity, one of the two main branches of the immune system. Analysis of the Staphylococcus pseudintermedius secretome using liquid chromatography mass spectrometry guided by genomic data, was used to identify an S. pseudintermedius exotoxin provisionally named SpEX. This exoprotein has low overall amino acid identity with the Staphylococcus aureus group of proteins named staphylococcal superantigen like proteins (SSLs) and staphylococcal enterotoxin- like toxin X (SEIX), but predictive modeling showed that it shares similar folds and domain architecture to these important virulence factors. In this study, we found SpEX binds to complement component C5, prevents complement mediated lysis of sensitized bovine red blood cells, kills polymorphonuclear leukocytes and monocytes and inhibits neutrophil migration at sub-lethal concentrations. A mutant version of SpEX, produced through amino acid substitution at selected positions, had diminished cytotoxicity. Anti-SpEX produced in dogs reduced the inhibitory effect of native SpEX on canine neutrophil migration and protected immune cells from the toxic effects of the native recombinant protein. These results suggest that SpEX likely plays an important role in S. pseudintermedius virulence and that attenuated SpEX may be an important candidate for inclusion in a vaccine against S. pseudintermedius infections.

View Accepted Manuscript (DOE)

Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1561612

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 7 Vol. 14; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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