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

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

Journal Article · 29 July 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.

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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, Vol. 14, Issue 7; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 4 works

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References (62)

Structural and Functional Properties of Staphylococcal Superantigen-Like Protein 4 Hermans, Stefan J.; Baker, Heather M.; Sequeira, Richard P. Infection and Immunity, Vol. 80, Issue 11 https://doi.org/10.1128/IAI.00764-12	journal	September 2012
Regulation of Staphylococcal Superantigen-Like Gene, ssl8, Expression in Staphylococcus aureus strain, RN6390 Pantrangi, M.; Singh, V. K.; Shukla, S. K. Clinical Medicine & Research, Vol. 13, Issue 1 https://doi.org/10.3121/cmr.2014.1226	journal	June 2014
The Three-dimensional Structure of a Superantigen-like Protein, SET3, from a Pathogenicity Island of the Staphylococcus aureus Genome Arcus, Vickery L.; Langley, Ries; Proft, Thomas Journal of Biological Chemistry, Vol. 277, Issue 35 https://doi.org/10.1074/jbc.M203914200	journal	June 2002
Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data Kearse, M.; Moir, R.; Wilson, A. Bioinformatics, Vol. 28, Issue 12 https://doi.org/10.1093/bioinformatics/bts199	journal	April 2012
3DLigandSite: predicting ligand-binding sites using similar structures Wass, Mark N.; Kelley, Lawrence A.; Sternberg, Michael J. E. Nucleic Acids Research, Vol. 38, Issue suppl_2 https://doi.org/10.1093/nar/gkq406	journal	May 2010
Identification of LukPQ, a novel, equid-adapted leukocidin of Staphylococcus aureus. Koop, Gerrit; Vrieling, Manouk; Storisteanu, Daniel ML Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.24528	text	January 2017
Antimicrobial resistance in methicillin susceptible and methicillin resistant Staphylococcus pseudintermedius of canine origin: Literature review from 1980 to 2013 Moodley, Arshnee; Damborg, Peter; Nielsen, Søren Saxmose Veterinary Microbiology, Vol. 171, Issue 3-4 https://doi.org/10.1016/j.vetmic.2014.02.008	journal	July 2014
The bacterial superantigen and superantigen-like proteins Fraser, John D.; Proft, Thomas Immunological Reviews, Vol. 225, Issue 1 https://doi.org/10.1111/j.1600-065X.2008.00681.x	journal	October 2008
The Staphylococcus intermedius group of bacterial pathogens: species re-classification, pathogenesis and the emergence of meticillin resistance: Recent studies of the Staphylococcus intermedius group Ross Fitzgerald, J. Veterinary Dermatology, Vol. 20, Issue 5-6 https://doi.org/10.1111/j.1365-3164.2009.00828.x	journal	October 2009
The Staphylococcus aureus superantigen SElX is a bifunctional toxin that inhibits neutrophil function Tuffs, Stephen W.; James, David B. A.; Bestebroer, Jovanka PLOS Pathogens, Vol. 13, Issue 9 https://doi.org/10.1371/journal.ppat.1006461	journal	September 2017
Inferring Reasons for the Failure of Staphylococcus aureus Vaccines in Clinical Trials Bagnoli, Fabio; Bertholet, Sylvie; Grandi, Guido Frontiers in Cellular and Infection Microbiology, Vol. 2 https://doi.org/10.3389/fcimb.2012.00016	journal	January 2012
Methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in veterinary medicine Weese, J. Scott; van Duijkeren, Engeline Veterinary Microbiology, Vol. 140, Issue 3-4 https://doi.org/10.1016/j.vetmic.2009.01.039	journal	January 2010
Clonal Complexes and Antimicrobial Susceptibility Profiles of Staphylococcus pseudintermedius Isolates from Dogs in the United States Videla, Ricardo; Solyman, Samar M.; Brahmbhatt, Akshar Microbial Drug Resistance, Vol. 24, Issue 1 https://doi.org/10.1089/mdr.2016.0250	journal	January 2018
Status of vaccine research and development of vaccines for Staphylococcus aureus Giersing, Birgitte K.; Dastgheyb, Sana S.; Modjarrad, Kayvon Vaccine, Vol. 34, Issue 26 https://doi.org/10.1016/j.vaccine.2016.03.110	journal	June 2016
Characterization of a leukocidin identified in Staphylococcus pseudintermedius Abouelkhair, Mohamed A.; Bemis, David A.; Giannone, Richard J. PLOS ONE, Vol. 13, Issue 9 https://doi.org/10.1371/journal.pone.0204450	journal	September 2018
Neutrophil integrin affinity regulation in adhesion, migration, and bacterial clearance Langereis, Jeroen D. Cell Adhesion & Migration, Vol. 7, Issue 6 https://doi.org/10.4161/cam.27293	journal	November 2013
SignalP 4.0: discriminating signal peptides from transmembrane regions Petersen, Thomas Nordahl; Brunak, Søren; von Heijne, Gunnar Nature Methods, Vol. 8, Issue 10 https://doi.org/10.1038/nmeth.1701	journal	September 2011
SA4Ag, a 4-antigen Staphylococcus aureus vaccine, rapidly induces high levels of bacteria-killing antibodies Begier, Elizabeth; Seiden, David Joshua; Patton, Michael Vaccine, Vol. 35, Issue 8 https://doi.org/10.1016/j.vaccine.2017.01.024	journal	February 2017
The Phyre2 web portal for protein modeling, prediction and analysis Kelley, Lawrence A.; Mezulis, Stefans; Yates, Christopher M. Nature Protocols, Vol. 10, Issue 6 https://doi.org/10.1038/nprot.2015.053	journal	May 2015
Identification of LukPQ, a novel, equid-adapted leukocidin of Staphylococcus aureus Koop, Gerrit; Vrieling, Manouk; Storisteanu, Daniel M. L. Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/srep40660	journal	January 2017
Manipulation of Innate and Adaptive Immunity by Staphylococcal Superantigens Tuffs, Stephen; Haeryfar, S.; McCormick, John Pathogens, Vol. 7, Issue 2 https://doi.org/10.3390/pathogens7020053	journal	May 2018
Structural basis for inhibition of TLR2 by staphylococcal superantigen-like protein 3 (SSL3) Koymans, Kirsten J.; Feitsma, Louris J.; Brondijk, T. Harma C. Proceedings of the National Academy of Sciences, Vol. 112, Issue 35 https://doi.org/10.1073/pnas.1502026112	journal	August 2015
The membrane proteome of male gametophyte in Solanum lycopersicum Paul, Puneet; Chaturvedi, Palak; Selymesi, Mario Journal of Proteomics, Vol. 131 https://doi.org/10.1016/j.jprot.2015.10.009	journal	January 2016
Complete Genome Sequence of Escherichia coli Bacteriophage PGT2 Golomidova, Alla K.; Kulikov, Eugene E.; Kudryavtseva, Anna V. Genome Announcements, Vol. 6, Issue 3 https://doi.org/10.1128/genomeA.01370-17	journal	January 2018
The potential use of toxin antibodies as a strategy for controlling acute Staphylococcus aureus infections Cheung, Gordon YC; Otto, Michael Expert Opinion on Therapeutic Targets, Vol. 16, Issue 6 https://doi.org/10.1517/14728222.2012.682573	journal	April 2012
Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance Magiorakos, A. -P.; Srinivasan, A.; Carey, R. B. Clinical Microbiology and Infection, Vol. 18, Issue 3 https://doi.org/10.1111/j.1469-0691.2011.03570.x	journal	March 2012
Staphylococcal enterotoxin-like X (SElX) is a unique superantigen with functional features of two major families of staphylococcal virulence factors Langley, Ries J.; Ting, Yi Tian; Clow, Fiona PLOS Pathogens, Vol. 13, Issue 9 https://doi.org/10.1371/journal.ppat.1006549	journal	September 2017
Progress Toward a Staphylococcus aureus Vaccine Daum, R. S.; Spellberg, B. Clinical Infectious Diseases, Vol. 54, Issue 4 https://doi.org/10.1093/cid/cir828	journal	December 2011
Crystal Structures of the Staphylococcal Toxin SSL5 in Complex with Sialyl Lewis X Reveal a Conserved Binding Site that Shares Common Features with Viral and Bacterial Sialic Acid Binding Proteins Baker, Heather M.; Basu, Indira; Chung, Matthew C. Journal of Molecular Biology, Vol. 374, Issue 5 https://doi.org/10.1016/j.jmb.2007.09.091	journal	December 2007
Staphylococcus pseudintermedius sp. nov., a coagulase-positive species from animals Devriese, L. A. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, Vol. 55, Issue 4 https://doi.org/10.1099/ijs.0.63413-0	journal	July 2005
Complete Genome Sequence of Staphylococcus pseudintermedius Type Strain LMG 22219 Abouelkhair, Mohamed A.; Riley, Matthew C.; Bemis, David A. Genome Announcements, Vol. 5, Issue 7 https://doi.org/10.1128/genomeA.01651-16	journal	February 2017
Neutrophils and Bacterial Immune Evasion Kobayashi, Scott D.; Malachowa, Natalia; DeLeo, Frank R. Journal of Innate Immunity, Vol. 10, Issue 5-6 https://doi.org/10.1159/000487756	journal	January 2018
Review on methicillin-resistant Staphylococcus pseudintermedius van Duijkeren, E.; Catry, B.; Greko, C. Journal of Antimicrobial Chemotherapy, Vol. 66, Issue 12 https://doi.org/10.1093/jac/dkr367	journal	September 2011
Fine-scale population structure and evidence for local adaptation in Australian giant black tiger shrimp (Penaeus monodon) using SNP analysis Vu, Nga T. T.; Zenger, Kyall R.; Guppy, Jarrod L. BMC Genomics, Vol. 21, Issue 1 https://doi.org/10.1186/s12864-020-07084-x	journal	September 2020
Multilocus Sequence Typing for Characterization of Staphylococcus pseudintermedius Solyman, S. M.; Black, C. C.; Duim, B. Journal of Clinical Microbiology, Vol. 51, Issue 1 https://doi.org/10.1128/JCM.02421-12	journal	October 2012
Complete Genome Sequences of Three Important Methicillin-Resistant Clinical Isolates of Staphylococcus pseudintermedius Riley, Matthew C.; Perreten, Vincent; Bemis, David A. Genome Announcements, Vol. 4, Issue 5 https://doi.org/10.1128/genomeA.01194-16	journal	October 2016
The crystal structure of staphylococcal superantigen-like protein 11 in complex with sialyl Lewis X reveals the mechanism for cell binding and immune inhibition: SSL11 binding to sialyl Lewis X Chung, Matthew C.; Wines, Bruce D.; Baker, Heather Molecular Microbiology, Vol. 66, Issue 6 https://doi.org/10.1111/j.1365-2958.2007.05989.x	journal	October 2007
Multilocus sequence typing for characterization of Staphylococcus pseudintermedius Eberlein, L. C.; Videla, R.; Wagenaar, J. A. American Society for Microbiology https://doi.org/10.7892/boris.43831	text	January 2013
Complete Genome Sequences of Three Important Methicillin-Resistant Clinical Isolates of Staphylococcus pseudintermedius. Perreten, Vincent; Kania, Stephen A.; Riley, Matthew C. American Society for Microbiology https://doi.org/10.7892/boris.95366	text	January 2016
Crystal Structures of the Staphylococcal Toxin SSL5 in Complex with Sialyl Lewis X Reveal a Conserved Binding Site that Shares Common Features with Viral and Bacterial Sialic Acid Binding Proteins Baker, Heather M.; Basu, Indira; Chung, Matthew C. Journal of Molecular Biology, Vol. 374, Issue 5 https://doi.org/10.1016/j.jmb.2007.09.091	journal	December 2007
The membrane proteome of male gametophyte in Solanum lycopersicum Paul, Puneet; Chaturvedi, Palak; Selymesi, Mario Journal of Proteomics, Vol. 131 https://doi.org/10.1016/j.jprot.2015.10.009	journal	January 2016
Status of vaccine research and development of vaccines for Staphylococcus aureus Giersing, Birgitte K.; Dastgheyb, Sana S.; Modjarrad, Kayvon Vaccine, Vol. 34, Issue 26 https://doi.org/10.1016/j.vaccine.2016.03.110	journal	June 2016
SignalP 4.0: discriminating signal peptides from transmembrane regions Petersen, Thomas Nordahl; Brunak, Søren; von Heijne, Gunnar Nature Methods, Vol. 8, Issue 10 https://doi.org/10.1038/nmeth.1701	journal	September 2011
The Phyre2 web portal for protein modeling, prediction and analysis Kelley, Lawrence A.; Mezulis, Stefans; Yates, Christopher M. Nature Protocols, Vol. 10, Issue 6 https://doi.org/10.1038/nprot.2015.053	journal	May 2015
Expression and function of protein A in Staphylococcus pseudintermedius Balachandran, Manasi; Bemis, David A.; Kania, Stephen A. Virulence, Vol. 9, Issue 1 https://doi.org/10.1080/21505594.2017.1403710	journal	March 2018
Progress Toward a Staphylococcus aureus Vaccine Daum, R. S.; Spellberg, B. Clinical Infectious Diseases, Vol. 54, Issue 4 https://doi.org/10.1093/cid/cir828	journal	December 2011
Staphylococcus pseudintermedius sp. nov., a coagulase-positive species from animals Devriese, L. A. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, Vol. 55, Issue 4 https://doi.org/10.1099/ijs.0.63413-0	journal	July 2005
Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance Magiorakos, A. -P.; Srinivasan, A.; Carey, R. B. Clinical Microbiology and Infection, Vol. 18, Issue 3 https://doi.org/10.1111/j.1469-0691.2011.03570.x	journal	March 2012
Complete Genome Sequences of Three Staphylococcus pseudintermedius Strains Isolated from Botswana Abouelkhair, Mohamed A.; Thompson, Riley; Riley, Matthew C. Genome Announcements, Vol. 6, Issue 10 https://doi.org/10.1128/genomea.01599-17	journal	March 2018
Identification, Cloning, and Characterization of Staphylococcus pseudintermedius Coagulase Sewid, Alaa H.; Hassan, M. Nabil; Ammar, A. M. Infection and Immunity, Vol. 86, Issue 8 https://doi.org/10.1128/iai.00027-18	journal	June 2018
Characterization of canine neutrophil granules O'Donnell, R. T.; Andersen, B. R. Infection and Immunity, Vol. 38, Issue 1 https://doi.org/10.1128/iai.38.1.351-359.1982	journal	October 1982
Complete Genome Sequence of the Canine Pathogen Staphylococcus pseudintermedius Zakour, N. L. B.; Bannoehr, J.; van den Broek, A. H. M. Journal of Bacteriology, Vol. 193, Issue 9 https://doi.org/10.1128/jb.00137-11	journal	March 2011
Characterization of a leukocidin identified in Staphylococcus pseudintermedius Abouelkhair, Mohamed A.; Bemis, David A.; Giannone, Richard J. PLOS ONE, Vol. 13, Issue 9 https://doi.org/10.1371/journal.pone.0204450	journal	September 2018
Staphylococcal enterotoxin-like X (SElX) is a unique superantigen with functional features of two major families of staphylococcal virulence factors Langley, Ries J.; Ting, Yi Tian; Clow, Fiona PLOS Pathogens, Vol. 13, Issue 9 https://doi.org/10.1371/journal.ppat.1006549	journal	September 2017
The potential use of toxin antibodies as a strategy for controlling acute Staphylococcus aureus infections Cheung, Gordon YC; Otto, Michael Expert Opinion on Therapeutic Targets, Vol. 16, Issue 6 https://doi.org/10.1517/14728222.2012.682573	journal	April 2012
Identification of LukPQ, a novel, equid-adapted leukocidin of Staphylococcus aureus. Koop, Gerrit; Vrieling, Manouk; Storisteanu, Daniel ML Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.24528	text	January 2017
Detección de Staphylococcus pseudintermedius resistentes a meticilina y a otros antimicrobianos de uso habitual en la clínica en piodermias caninas Giacoboni, Gabriela; Vinocur, Florencia; Fauret, Natali Analecta Veterinaria, Vol. 37, Issue 2 https://doi.org/10.24215/15142590e012	journal	December 2017
Inferring Reasons for the Failure of Staphylococcus aureus Vaccines in Clinical Trials Bagnoli, Fabio; Bertholet, Sylvie; Grandi, Guido Frontiers in Cellular and Infection Microbiology, Vol. 2 https://doi.org/10.3389/fcimb.2012.00016	journal	January 2012
Manipulation of Innate and Adaptive Immunity by Staphylococcal Superantigens Tuffs, Stephen; Haeryfar, S.; McCormick, John Pathogens, Vol. 7, Issue 2 https://doi.org/10.3390/pathogens7020053	journal	May 2018
In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration Kusek, Mark E.; Pazos, Michael A.; Pirzai, Waheed Journal of Visualized Experiments, Issue 83 https://doi.org/10.3791/50823	journal	January 2014
Neutrophil integrin affinity regulation in adhesion, migration, and bacterial clearance Langereis, Jeroen D. Cell Adhesion & Migration, Vol. 7, Issue 6 https://doi.org/10.4161/cam.27293	journal	November 2013
Characterization of recombinant wild-type and nontoxigenic protein A from Staphylococcus pseudintermedius Abouelkhair, Mohamed A.; Bemis, David A.; Kania, Stephen A. Taylor & Francis https://doi.org/10.6084/m9.figshare.6886583.v1	text	January 2018