Journal of Molecular Medicine

, Volume 92, Issue 2, pp 139–149

Nafcillin enhances innate immune-mediated killing of methicillin-resistant Staphylococcus aureus

  • George Sakoulas
  • Cheryl Y. Okumura
  • Wdee Thienphrapa
  • Joshua Olson
  • Poochit Nonejuie
  • Quang Dam
  • Abhay Dhand
  • Joseph Pogliano
  • Michael R. Yeaman
  • Mary E. Hensler
  • Arnold S. Bayer
  • Victor Nizet
Original Article

Abstract

Based on in vitro synergy studies, the addition of nafcillin to daptomycin was used to treat refractory methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. Daptomycin is a de facto cationic antimicrobial peptide in vivo, with antistaphylococcal mechanisms reminiscent of innate host defense peptides (HDPs). In this study, the effects of nafcillin on HDP activity against MRSA were examined in vitro and in vivo. Exposures to β-lactam antimicrobials in general, and nafcillin in particular, significantly increased killing of S. aureus by selected HDPs from keratinocytes, neutrophils, and platelets. This finding correlated with enhanced killing of MRSA by whole blood, neutrophils, and keratinocytes after growth in nafcillin. Finally, nafcillin pretreatment ex vivo reduced MRSA virulence in a murine subcutaneous infection model. Despite the lack of direct activity against MRSA, these studies show potent, consistent, and generalized nafcillin-mediated “sensitization” to increased killing of MRSA by various components of the innate host response. The use of nafcillin as adjunctive therapy in MRSA bacteremia merits further study and should be considered in cases refractory to standard therapy.

Key messages

  • Nafcillin has been used as adjunctive therapy to clear persistent MRSA bacteremia.

  • Nafcillin enhances killing of MRSA by a cadre of innate host defense peptides.

  • Nafcillin increases binding of human cathelicidin LL-37 to the MRSA membrane.

  • Nafcillin enhances killing of MRSA by neutrophils.

  • Nafcillin reduces virulence of MRSA in a murine subcutaneous infection model.

Keywords

MRSA Innate immunity Beta-lactams Nafcillin Host defense peptides 

References

  1. 1.
    Grundmann H, Aires-de-Sousa M, Boyce J, Tiemersma E (2006) Emergence and resurgence of methicillin-resistant Staphylococcus aureus as a public-health threat. Lancet 368:874–885PubMedCrossRefGoogle Scholar
  2. 2.
    Mediavilla JR, Chen L, Mathema B, Kreiswirth BN (2012) Global epidemiology of community-associated methicillin resistant Staphylococcus aureus (CA-MRSA). Curr Opin Microbiol 15:588–595PubMedCrossRefGoogle Scholar
  3. 3.
    Chambers HF, Deleo FR (2009) Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol 7:629–641PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Como-Sabetti K, Harriman KH, Buck JM, Glennen A, Boxrud DJ, Lynfield R (2009) Community-associated methicillin-resistant Staphylococcus aureus: trends in case and isolate characteristics from six years of prospective surveillance. Public Health Rep 124:427–435PubMedCentralPubMedGoogle Scholar
  5. 5.
    DeLeo FR, Otto M, Kreiswirth BN, Chambers HF (2010) Community-associated methicillin-resistant Staphylococcus aureus. Lancet 375:1557–1568PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Cosgrove SE, Qi Y, Kaye KS, Harbarth S, Karchmer AW, Carmeli Y (2005) The impact of methicillin resistance in Staphylococcus aureus bacteremia on patient outcomes: mortality, length of stay, and hospital charges. Infect Cont Hosp Epidemiol 26:166–174CrossRefGoogle Scholar
  7. 7.
    Reed SD, Friedman JY, Engemann JJ, Griffiths RI, Anstrom KJ, Kaye KS, Stryjewski ME, Szczech LA, Reller LB, Corey GR et al (2005) Costs and outcomes among hemodialysis-dependent patients with methicillin-resistant or methicillin-susceptible Staphylococcus aureus bacteremia. Infect Cont Hosp Epidemiol 26:175–183CrossRefGoogle Scholar
  8. 8.
    Sakoulas G, Moise-Broder PA, Schentag J, Forrest A, Moellering RC Jr, Eliopoulos GM (2004) Relationship of MIC and bactericidal activity to efficacy of vancomycin for treatment of methicillin-resistant Staphylococcus aureus bacteremia. J Clin Microbiol 42:2398–2402PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Soriano A, Marco F, Martinez JA, Pisos E, Almela M, Dimova VP, Alamo D, Ortega M, Lopez J, Mensa J (2008) Influence of vancomycin minimum inhibitory concentration on the treatment of methicillin-resistant Staphylococcus aureus bacteremia. Clin Infect Dis 46:193–200PubMedCrossRefGoogle Scholar
  10. 10.
    Otto M (2010) Basis of virulence in community-associated methicillin-resistant Staphylococcus aureus. Ann Rev Microbiol 64:143–162CrossRefGoogle Scholar
  11. 11.
    David MZ, Daum RS (2010) Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic. Clin Microbiol Rev 23:616–687PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Watkins RR, David MZ, Salata RA (2012) Current concepts on the virulence mechanisms of methicillin-resistant Staphylococcus aureus. J Med Microbiol 61:1179–1193PubMedCrossRefGoogle Scholar
  13. 13.
    Cederlund A, Gudmundsson GH, Agerberth B (2011) Antimicrobial peptides important in innate immunity. FEBS J 278:3942–3951PubMedCrossRefGoogle Scholar
  14. 14.
    Gallo RL, Hooper LV (2012) Epithelial antimicrobial defence of the skin and intestine. Nat Rev Immunol 12:503–516PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Peschel A, Collins LV (2001) Staphylococcal resistance to antimicrobial peptides of mammalian and bacterial origin. Peptides 22:1651–1659PubMedCrossRefGoogle Scholar
  16. 16.
    Kraus D, Peschel A (2008) Staphylococcus aureus evasion of innate antimicrobial defense. Future Microbiol 3:437–451PubMedCrossRefGoogle Scholar
  17. 17.
    Clarke SR, Mohamed R, Bian L, Routh AF, Kokai-Kun JF, Mond JJ, Tarkowski A, Foster SJ (2007) The Staphylococcus aureus surface protein IsdA mediates resistance to innate defenses of human skin. Cell Host Microbe 1:199–212PubMedCrossRefGoogle Scholar
  18. 18.
    Nizet V (2006) Antimicrobial peptide resistance mechanisms of human bacterial pathogens. Curr Issues Mol Biol 8:11–26PubMedGoogle Scholar
  19. 19.
    Midorikawa K, Ouhara K, Komatsuzawa H, Kawai T, Yamada S, Fujiwara T, Yamazaki K, Sayama K, Taubman MA, Kurihara H et al (2003) Staphylococcus aureus susceptibility to innate antimicrobial peptides, beta-defensins and CAP18, expressed by human keratinocytes. Infect Immun 71:3730–3739PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Ouhara K, Komatsuzawa H, Kawai T, Nishi H, Fujiwara T, Fujiue Y, Kuwabara M, Sayama K, Hashimoto K, Sugai M (2008) Increased resistance to cationic antimicrobial peptide LL-37 in methicillin-resistant strains of Staphylococcus aureus. J Antimicrob Chemother 61:1266–1269PubMedCrossRefGoogle Scholar
  21. 21.
    Liu C, Bayer A, Cosgrove SE, Daum RS, Fridkin SK, Gorwitz RJ, Kaplan SL, Karchmer AW, Levine DP, Murray BE et al (2011) Clinical practice guidelines by the Infectious Diseases Society of America for the treatment of methicillin-resistant Staphylococcus aureus infections in adults and children. Clin Infect Dis 52:e18–e55PubMedCrossRefGoogle Scholar
  22. 22.
    Dhand A, Bayer AS, Pogliano J, Yang SJ, Bolaris M, Nizet V, Wang G, Sakoulas G (2011) Use of antistaphylococcal beta-lactams to increase daptomycin activity in eradicating persistent bacteremia due to methicillin-resistant Staphylococcus aureus: role of enhanced daptomycin binding. Clin Infect Dis 53:158–163PubMedCrossRefGoogle Scholar
  23. 23.
    Bayer AS, Schneider T, Sahl HG (2013) Mechanisms of daptomycin resistance in Staphylococcus aureus: role of the cell membrane and cell wall. Ann NY Acad Sci 1277:139–158PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Vilhena C, Bettencourt A (2012) Daptomycin: a review of properties, clinical use, drug delivery and resistance. Mini-Rev Med Chem 12:202–209PubMedCrossRefGoogle Scholar
  25. 25.
    Tedesco KL, Rybak MJ (2004) Daptomycin. Pharmacotherapy 24:41–57PubMedCrossRefGoogle Scholar
  26. 26.
    Mishra NN, Bayer AS, Moise PA, Yeaman MR, Sakoulas G (2012) Reduced susceptibility to host-defense cationic peptides and daptomycin coemerge in methicillin-resistant Staphylococcus aureus from daptomycin-naive bacteremic patients. J Infect Dis 206:1160–1167PubMedCrossRefGoogle Scholar
  27. 27.
    Mishra NN, McKinnell J, Yeaman MR, Rubio A, Nast CC, Chen L, Kreiswirth BN, Bayer AS (2011) In vitro cross-resistance to daptomycin and host defense cationic antimicrobial peptides in clinical methicillin-resistant Staphylococcus aureus isolates. Antimicrob Agents Chemother 55:4012–4018PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Xiong YQ, Bayer AS, Elazegui L, Yeaman MR (2006) A synthetic congener modeled on a microbicidal domain of thrombin-induced platelet microbicidal protein 1 recapitulates staphylocidal mechanisms of the native molecule. Antimicrob Agents Chemother 50:3786–3792PubMedCentralPubMedCrossRefGoogle Scholar
  29. 29.
    Mollnes TE, Brekke OL, Fung M, Fure H, Christiansen D, Bergseth G, Videm V, Lappegard KT, Kohl J, Lambris JD (2002) Essential role of the C5a receptor in E. coli-induced oxidative burst and phagocytosis revealed by a novel lepirudin-based human whole blood model of inflammation. Blood 100:1869–1877PubMedGoogle Scholar
  30. 30.
    Kristian SA, Datta V, Weidenmaier C, Kansal R, Fedtke I, Peschel A, Gallo RL, Nizet V (2005) D-alanylation of teichoic acids promotes group A Streptococcus antimicrobial peptide resistance, neutrophil survival, and epithelial cell invasion. J Bacteriol 187:6719–6725PubMedCentralPubMedCrossRefGoogle Scholar
  31. 31.
    von Kockritz-Blickwede M, Chow OA, Nizet V (2009) Fetal calf serum contains heat-stable nucleases that degrade neutrophil extracellular traps. Blood 114:5245–5246CrossRefGoogle Scholar
  32. 32.
    Schaller-Bals S, Schulze A, Bals R (2002) Increased levels of antimicrobial peptides in tracheal aspirates of newborn infants during infection. Am J Respir Crit Care Med 165:992–995PubMedCrossRefGoogle Scholar
  33. 33.
    Chen CI, Schaller-Bals S, Paul KP, Wahn U, Bals R (2004) Beta-defensins and LL-37 in bronchoalveolar lavage fluid of patients with cystic fibrosis. J Cyst Fibros 3:45–50PubMedCrossRefGoogle Scholar
  34. 34.
    Ong PY, Ohtake T, Brandt C, Strickland I, Boguniewicz M, Ganz T, Gallo RL, Leung DY (2002) Endogenous antimicrobial peptides and skin infections in atopic dermatitis. N Engl J Med 347:1151–1160PubMedCrossRefGoogle Scholar
  35. 35.
    Hosokawa I, Hosokawa Y, Komatsuzawa H, Goncalves RB, Karimbux N, Napimoga MH, Seki M, Ouhara K, Sugai M, Taubman MA et al (2006) Innate immune peptide LL-37 displays distinct expression pattern from beta-defensins in inflamed gingival tissue. Clin Exp Immunol 146:218–225PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Holden MT, Feil EJ, Lindsay JA, Peacock SJ, Day NP, Enright MC, Foster TJ, Moore CE, Hurst L, Atkin R et al (2004) Complete genomes of two clinical Staphylococcus aureus strains: evidence for the rapid evolution of virulence and drug resistance. Proc Natl Acad Sci U S A 101:9786–9791PubMedCentralPubMedCrossRefGoogle Scholar
  37. 37.
    Nizet V, Ohtake T, Lauth X, Trowbridge J, Rudisill J, Dorschner RA, Pestonjamasp V, Piraino J, Huttner K, Gallo RL (2001) Innate antimicrobial peptide protects the skin from invasive bacterial infection. Nature 414:454–457PubMedCrossRefGoogle Scholar
  38. 38.
    Sorensen O, Arnljots K, Cowland JB, Bainton DF, Borregaard N (1997) The human antibacterial cathelicidin, hCAP-18, is synthesized in myelocytes and metamyelocytes and localized to specific granules in neutrophils. Blood 90:2796–2803PubMedGoogle Scholar
  39. 39.
    Faurschou M, Sorensen OE, Johnsen AH, Askaa J, Borregaard N (2002) Defensin-rich granules of human neutrophils: characterization of secretory properties. Biochim Biophysica Acta 1591:29–35CrossRefGoogle Scholar
  40. 40.
    Yeaman MR (1997) The role of platelets in antimicrobial host defense. Clin Infect Dis 25:951–968PubMedCrossRefGoogle Scholar
  41. 41.
    Yeaman MR, Bayer AS (2006) Antimicrobial peptides versus invasive infections. Curr Top Microbiol Immunol 306:111–152PubMedGoogle Scholar
  42. 42.
    Jann NJ, Schmaler M, Kristian SA, Radek KA, Gallo RL, Nizet V, Peschel A, Landmann R (2009) Neutrophil antimicrobial defense against Staphylococcus aureus is mediated by phagolysosomal but not extracellular trap-associated cathelicidin. J Leuk Biol 86:1159–1169CrossRefGoogle Scholar
  43. 43.
    Braff MH, Zaiou M, Fierer J, Nizet V, Gallo RL (2005) Keratinocyte production of cathelicidin provides direct activity against bacterial skin pathogens. Infect Immun 73:6771–6781PubMedCentralPubMedCrossRefGoogle Scholar
  44. 44.
    Levy SB, Marshall B (2004) Antibacterial resistance worldwide: causes, challenges and responses. Nat Med 10:S122–S129PubMedCrossRefGoogle Scholar
  45. 45.
    Spellberg B (2008) Antibiotic resistance and antibiotic development. Lancet Infect Dis 8:211–212PubMedCrossRefGoogle Scholar
  46. 46.
    Spellberg B, Blaser M, Guidos RJ, Boucher HW, Bradley JS, Eisenstein BI, Gerding D, Lynfield R, Reller LB, Rex J et al (2011) Combating antimicrobial resistance: policy recommendations to save lives. Clin Infect Dis 52(Suppl 5):S397–S428PubMedGoogle Scholar
  47. 47.
    Klevens RM, Morrison MA, Nadle J, Petit S, Gershman K, Ray S, Harrison LH, Lynfield R, Dumyati G, Townes JM et al (2007) Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA 298:1763–1771PubMedCrossRefGoogle Scholar
  48. 48.
    Chang FY, Peacock JE Jr, Musher DM, Triplett P, MacDonald BB, Mylotte JM, O’Donnell A, Wagener MM, Yu VL (2003) Staphylococcus aureus bacteremia: recurrence and the impact of antibiotic treatment in a prospective multicenter study. Medicine 82:333–339PubMedCrossRefGoogle Scholar
  49. 49.
    Schweizer ML, Furuno JP, Harris AD, Johnson JK, Shardell MD, McGregor JC, Thom KA, Cosgrove SE, Sakoulas G, Perencevich EN (2011) Comparative effectiveness of nafcillin or cefazolin versus vancomycin in methicillin-susceptible Staphylococcus aureus bacteremia. BMC Infect Dis 11:279PubMedCentralPubMedCrossRefGoogle Scholar
  50. 50.
    Sakoulas G, Bayer AS, Pogliano J, Tsuji BT, Yang SJ, Mishra NN, Nizet V, Yeaman MR, Moise PA (2012) Ampicillin enhances daptomycin- and cationic host defense peptide-mediated killing of ampicillin- and vancomycin-resistant Enterococcus faecium. Antimicrob Agents Chemother 56:838–844PubMedCentralPubMedCrossRefGoogle Scholar
  51. 51.
    al-Obeid S, Gutmann L, Williamson R (1990) Correlation of penicillin-induced lysis of Enterococcus faecium with saturation of essential penicillin-binding proteins and release of lipoteichoic acid. Antimicrob Agents Chemother 34:1901–1907PubMedCentralPubMedCrossRefGoogle Scholar
  52. 52.
    Pogliano J, Pogliano N, Silverman JA (2012) Daptomycin-mediated reorganization of membrane architecture causes mislocalization of essential cell division proteins. J Bacteriol 194:4494–4504PubMedCentralPubMedCrossRefGoogle Scholar
  53. 53.
    McGillivray SM, Tran DN, Ramadoss NS, Alumasa JN, Okumura CY, Sakoulas G, Vaughn MM, Zhang DX, Keiler KC, Nizet V (2012) Pharmacological inhibition of the ClpXP protease increases bacterial susceptibility to host cathelicidin antimicrobial peptides and cell envelope-active antibiotics. Antimicrob Agents Chemother 56:1854–1861PubMedCentralPubMedCrossRefGoogle Scholar
  54. 54.
    Tan CM, Therien AG, Lu J, Lee SH, Caron A, Gill CJ, Lebeau-Jacob C, Benton-Perdomo L, Monteiro JM, Pereira PM, et al (2012) Restoring methicillin-resistant Staphylococcus aureus susceptibility to beta-lactam antibiotics. Science Trans Med 4: 126ra135Google Scholar
  55. 55.
    Crawford T, Rodvold KA, Solomkin JS (2012) Vancomycin for surgical prophylaxis? Clin Infect Dis 54: 1474–1479Google Scholar
  56. 56.
    Yeaman MR, Norman DC, Bayer AS (1992) Platelet microbicidal protein enhances antibiotic-induced killing of and postantibiotic effect in Staphylococcus aureus. Antimicrob Agents Chemother 36:1665–1670PubMedCentralPubMedCrossRefGoogle Scholar
  57. 57.
    Kristian SA, Timmer AM, Liu GY, Lauth X, Sal-Man N, Rosenfeld Y, Shai Y, Gallo RL, Nizet V (2007) Impairment of innate immune killing mechanisms by bacteriostatic antibiotics. FASEB J 21:1107–1116PubMedCrossRefGoogle Scholar
  58. 58.
    Mehta S, Singh C, Plata KB, Chanda PK, Paul A, Riosa S, Rosato RR, Rosato AE (2012) Beta-lactams increase the antibacterial activity of daptomycin against clinical methicillin-resistant Staphylococcus aureus strains and prevent selection of daptomycin-resistant derivatives. Antimicrob Agents Chemother 56:6192–6200PubMedCentralPubMedCrossRefGoogle Scholar
  59. 59.
    Moise PA, Amodio-Groton M, Rashid M, Lamp KC, Hoffman-Roberts HL, Sakoulas G, Yoon MJ, Schweitzer S, Rastogi A (2013) Multicenter evaluation of the clinical outcomes of daptomycin with and without concomitant beta-lactams in patients with Staphylococcus aureus bacteremia and mild to moderate renal impairment. Antimicrob Agents Chemother 57:1192–1200PubMedCentralPubMedCrossRefGoogle Scholar
  60. 60.
    Stevens DL, Ma Y, Salmi DB, McIndoo E, Wallace RJ, Bryant AE (2007) Impact of antibiotics on expression of virulence-associated exotoxin genes in methicillin-sensitive and methicillin-resistant Staphylococcus aureus. J Infect Dis 195:202–211PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • George Sakoulas
    • 1
    • 7
  • Cheryl Y. Okumura
    • 1
    • 4
  • Wdee Thienphrapa
    • 1
  • Joshua Olson
    • 1
  • Poochit Nonejuie
    • 3
  • Quang Dam
    • 1
  • Abhay Dhand
    • 5
  • Joseph Pogliano
    • 3
  • Michael R. Yeaman
    • 6
  • Mary E. Hensler
    • 1
  • Arnold S. Bayer
    • 6
  • Victor Nizet
    • 1
    • 2
    • 7
  1. 1.Department of PediatricsUniversity of California, San DiegoLa JollaUSA
  2. 2.Skaggs School of Pharmacy & Pharmaceutical SciencesUniversity of California, San DiegoLa JollaUSA
  3. 3.Department of Biological SciencesUniversity of California, San DiegoLa JollaUSA
  4. 4.Department of BiologyOccidental CollegeLos AngelesUSA
  5. 5.Department of MedicineNew York Medical CollegeValhallaUSA
  6. 6.David Geffen School of Medicine at UCLA and the Los Angeles Biomedical Research Institute at Harbor–UCLA Medical CenterTorranceUSA
  7. 7.Division of Pediatric Pharmacology & Drug DiscoveryUniversity of California San Diego School of MedicineLa JollaUSA

Personalised recommendations