Nitrite modulates bacterial antibiotic susceptibility and biofilm formation in association with airway epithelial cells
Open accessAbstract
Pseudomonas aeruginosa is the major pathogenic bacteria in cystic fibrosis and other forms of bronchiectasis. Growth in antibiotic-resistant biofilms contributes to the virulence of this organism. Sodium nitrite has antimicrobial properties and has been tolerated as a nebulized compound at high concentrations in human subjects with pulmonary hypertension; however, its effects have not been evaluated on biotic biofilms or in combination with other clinically useful antibiotics. We grew P. aeruginosa on the apical surface of primary human airway epithelial cells to test the efficacy of sodium nitrite against biotic biofilms. Nitrite alone prevented 99% of biofilm growth. We then identified significant cooperative interactions between nitrite and polymyxins. For P. aeruginosa growing on primary CF airway cells, combining nitrite and colistimethate resulted in an additional log of bacterial inhibition compared to treating with either agent alone. Nitrite and colistimethate additively inhibited oxygen consumption by P. aeruginosa. Surprisingly, whereas the antimicrobial effects of nitrite in planktonic, aerated cultures are nitric oxide (NO) dependent, antimicrobial effects under other growth conditions are not. The inhibitory effect of nitrite on bacterial oxygen consumption and biofilm growth did not require NO as an intermediate as chemically scavenging NO did not block growth inhibition. These data suggest an NO-radical independent nitrosative or oxidative inhibition of respiration. The combination of nebulized sodium nitrite and colistimethate may provide a novel therapy for chronic P. aeruginosa airway infections, because sodium nitrite, unlike other antibiotic respiratory chain "poisons," can be safely nebulized at high concentration in humans.
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Nitrite can exert pH-dependent antimicrobial effects via formation of reactive secondary nitrogen oxides that can disrupt aerobic respiration, inhibit bacterial acid production, and disrupt biofilm formation.
“Nitrite can exert pH-dependent antimicrobial effects[204,232,233] via formation of reactive secondary nitrogen oxides that can disrupt aerobic respiration[234], inhibit bacterial acid production, and disrupt biofilm formation[235–237].”
From Enterosalivary nitrate metabolism and the microbiome: Intersection of microbial metabolism, nitric oxide and diet in cardiac and pulmonary vascular health (2017) · cites this paper as [236]
High salivary nitrite concentrations may exert antimicrobial effects elsewhere in the oral cavity and gut, possibly disrupting community composition or function of biologically beneficial populations of nitrate-reducing bacteria.
“Yet, despite these localized, periodontal effects on microbial populations, high salivary nitrite concentrations may also exert antimicrobial effects elsewhere in the oral cavity and gut, possibly disrupting community composition or function of biologically beneficial populations of nitrate-reducing bacteria[204,232–237].”
From Enterosalivary nitrate metabolism and the microbiome: Intersection of microbial metabolism, nitric oxide and diet in cardiac and pulmonary vascular health (2017) · cites this paper as [236]
Nitrite is currently under investigation as an adjunct to standard antibiotic therapy.
“Nitrite, for instance, disrupts protective biofilm formation by pathogenic bacteria and is currently under investigation as an adjunct to standard antibiotic therapy[235,236].”
From Enterosalivary nitrate metabolism and the microbiome: Intersection of microbial metabolism, nitric oxide and diet in cardiac and pulmonary vascular health (2017) · cites this paper as [236]
Nitrite disrupts protective biofilm formation by pathogenic bacteria.
“Nitrite, for instance, disrupts protective biofilm formation by pathogenic bacteria and is currently under investigation as an adjunct to standard antibiotic therapy[235,236].”
From Enterosalivary nitrate metabolism and the microbiome: Intersection of microbial metabolism, nitric oxide and diet in cardiac and pulmonary vascular health (2017) · cites this paper as [236]
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Cites
Wiegand, I., Hilpert, K., & Hancock, R. E. W. (2008) · Nature Protocols · doi:10.1038/nprot.2007.521 · Cited by 6,156 · see citing works
Quantification of biofilm structures by the novel computer program comstat
Heydorn, A., Nielsen, A. T., Hentzer, M., Sternberg, C., Givskov, M., Ersbøll, B. K., & Molin, S. (2000) · Microbiology · doi:10.1099/00221287-146-10-2395 · Cited by 2,185 · see citing works
Genetic adaptation by <i>Pseudomonas aeruginosa</i> to the airways of cystic fibrosis patients
Smith, E. E., Buckley, D. G., Wu, Z., Saenphimmachak, C., Hoffman, L. R., D’Argenio, D. A., Miller, S. I., Ramsey, B. W., Speert, D. P., Moskowitz, S. M., et al. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0602138103 · Cited by 1,319 · see citing works
Self-citationA genetic basis for Pseudomonas aeruginosa biofilm antibiotic resistance
Mah, T., Pitts, B., Pellock, B. J., Walker, G. C., Stewart, P. S., & O’Toole, G. A. (2003) · Nature · doi:10.1038/nature02122 · Cited by 1,190 · see citing works
Cardioprotection by S-nitrosation of a cysteine switch on mitochondrial complex I
Chouchani, E. T., Methner, C., Nadtochiy, S. M., Logan, A., Pell, V. R., Ding, S., James, A. M., Cochemé, H. M., Reinhold, J., Lilley, K. S., et al. (2013) · Nature Medicine · doi:10.1038/nm.3212 · Cited by 619 · cited 1× · see citing works
“In eukaryotes, mitochondrial complex I is a target for cysteine S-nitrosation, which confers protection from ischemia/perfusion injury (52, 53).”
BackgroundPseudomonas aeruginosa Anaerobic Respiration in Biofilms
Yoon, S. S., Hennigan, R. F., Hilliard, G. M., Ochsner, U. A., Parvatiyar, K., Kamani, M. C., Allen, H. L., Dekievit, T., Gardner, P. R., Schwab, U. E., et al. (2002) · Developmental Cell · doi:10.1016/s1534-5807(02)00295-2 · Cited by 587 · see citing works
Shiva, S., Sack, M. N., Greer, J. J., Duranski, M., Ringwood, L. A., Burwell, L., Wang, X., MacArthur, P. H., Shoja, A., Raghavachari, N., et al. (2007) · The Journal of Experimental Medicine · doi:10.1084/jem.20070198 · Cited by 555 · see citing works
Self-citationPamp, S. J., Gjermansen, M., Johansen, H. K., & Tolker‐Nielsen, T. (2008) · Molecular Microbiology · doi:10.1111/j.1365-2958.2008.06152.x · Cited by 486 · see citing works
Endogenous Nitric Oxide Protects Bacteria Against a Wide Spectrum of Antibiotics
Gusarov, I., Shatalin, K., Starodubtseva, M., & Nudler, E. (2009) · Science · doi:10.1126/science.1175439 · Cited by 417 · see citing works
Inhaled Colistin in Patients with Bronchiectasis and Chronic Pseudomonas aeruginosa Infection
HAWORTH, C. S., Foweraker, J. E., Wilkinson, P., Kenyon, R. F., & Bilton, D. (2014) · American Journal of Respiratory and Critical Care Medicine · doi:10.1164/rccm.201312-2208oc · Cited by 263 · see citing works
Reddy, D. S. A., Lancaster, J. R., & Cornforth, D. P. (1983) · Science · doi:10.1126/science.6308761 · Cited by 258 · see citing works
Miller, A. K., Brannon, M. K., Stevens, L., et al. (2011) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.05391-11 · Cited by 225 · see citing works
Hill, D., Rose, B. R., Pajkos, A., Robinson, M., Bye, P. T. P., Bell, S. C., Elkins, M. R., Thompson, B., MacLeod, C., Aaron, S. D., et al. (2005) · Journal of Clinical Microbiology · doi:10.1128/jcm.43.10.5085-5090.2005 · Cited by 213 · see citing works
Yoon, S. S. (2006) · Journal of Clinical Investigation · doi:10.1172/jci24684 · Cited by 212 · see citing works
Pharmacokinetics of inhaled colistin in patients with cystic fibrosis
Ratjen, F. A., Rietschel, E., Kasel, D., et al. (2006) · Journal of Antimicrobial Chemotherapy · doi:10.1093/jac/dki461 · Cited by 208 · see citing works
Schuster, A., Haliburn, C., Döring, G., Goldman, M. H., & for the Freedom Study Group (2012) · Thorax · doi:10.1136/thoraxjnl-2012-202059 · Cited by 206 · see citing works
Inhibition of staphylococcal biofilm formation by nitrite
Schlag, S., Nerz, C., Birkenstock, T. A., Altenberend, F., & Götz, F. (2007) · Journal of Bacteriology · doi:10.1128/jb.00598-07 · Cited by 196 · see citing works
Moreau‐Marquis, S., Bomberger, J. M., Anderson, G. G., Swiatecka‐Urban, A., Ye, S., O’Toole, G. A., & Stanton, B. A. (2008) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00391.2007 · Cited by 187 · see citing works
Self-citationRhee, K. Y., Erdjument‐Bromage, H., Tempst, P., & Nathan, C. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0406133102 · Cited by 167 · see citing works
Yapa, S. W. S., Li, J., Patel, K., Wilson, J. W., Dooley, M. J., George, J., Clark, D., Poole, S. G., Williams, E., Porter, C. J. H., et al. (2014) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.01705-13 · Cited by 162 · see citing works
Mechanisms of nitrite bioactivation
Kim-Shapiro, D. B., & Gladwin, M. T. (2014) · Nitric Oxide · doi:10.1016/j.niox.2013.11.002 · Cited by 157 · see citing works
Self-citationStages of Polymyxin B Interaction with the Escherichia coli Cell Envelope
Daugelavičius, R., Bakienė, E., & Bamford, D. H. (2000) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.44.11.2969-2978.2000 · Cited by 154 · see citing works
Schaible, B., Taylor, C. T., & Schaffer, K. (2012) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.05574-11 · Cited by 133 · see citing works
<i>Helicobacter pylori</i> is killed by nitrite under acidic conditions
Dykhuizen, R. S., Fraser, A., McKenzie, H., Golden, M., Leifert, C., & Benjamin, N. (1998) · Gut · doi:10.1136/gut.42.3.334 · Cited by 131 · see citing works
Spilker, T., Vandamme, P. A. R., & LiPuma, J. J. (2012) · Journal of Clinical Microbiology · doi:10.1128/jcm.00814-12 · Cited by 127 · see citing works
The Diversity of Microbial Responses to Nitric Oxide and Agents of Nitrosative Stress
Bowman, L. A. H., McLean, S., Poole, R. K., & Fukuto, J. M. (2011) · Advances in microbial physiology/Advances in Microbial Physiology · doi:10.1016/b978-0-12-387661-4.00006-9 · Cited by 117 · see citing works
Miller, C., McMullin, B., Ghaffari, A., Stenzler, A., Pick, N., Roscoe, D. L., Ghahary, A., ROAD, J. D., & Av‐Gay, Y. (2008) · Nitric Oxide · doi:10.1016/j.niox.2008.08.002 · Cited by 105 · see citing works
Nitrite inhibition of aerobic bacteria
Rowe, J. J., Yarbrough, J. M., Rake, J. B., & Eagon, R. G. (1979) · Current Microbiology · doi:10.1007/bf02601735 · Cited by 97 · see citing works
Braun, C., & Zumft, W. G. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)54420-3 · Cited by 94 · see citing works
Kolpen, M., Kühl, M., Bjarnsholt, T., Moser, C. E., Hansen, C. R., Liengaard, L., Kharazmi, A., Pressler, T., Høiby, N., & Jensen, P. Ø. (2014) · PLoS ONE · doi:10.1371/journal.pone.0084353 · Cited by 93 · see citing works
Nitric Oxide Evokes an Adaptive Response to Oxidative Stress by Arresting Respiration
Husain, M., Bourret, T. J., McCollister, B. D., Jones‐Carson, J., Laughlin, J. R., & Vázquez‐Torres, A. (2008) · Journal of Biological Chemistry · doi:10.1074/jbc.m708845200 · Cited by 89 · see citing works
Nitric oxide‐induced bacteriostasis and modification of iron‐sulphur proteins in Escherichia coli
Ren, B., Zhang, N., Yang, J., & Ding, H. (2008) · Molecular Microbiology · doi:10.1111/j.1365-2958.2008.06464.x · Cited by 80 · see citing works
LaPorte, D. C., Rosenthal, K. S., & Storm, D. R. (1977) · Biochemistry · doi:10.1021/bi00627a019 · Cited by 76 · see citing works
Chiang, W., Pamp, S. J., Nilsson, M., et al. (2012) · FEMS Immunology & Medical Microbiology · doi:10.1111/j.1574-695x.2012.00929.x · Cited by 71 · see citing works
Yoon, S. S., Karabulut, A., Lipscomb, J. D., Hennigan, R. F., Lymar, S. V., Groce, S. L., Herr, A. B., Howell, M. L., Kiley, P. J., Schurr, M. J., et al. (2007) · The EMBO Journal · doi:10.1038/sj.emboj.7601787 · Cited by 71 · see citing works
Bueno, M., Wang, J., Mora, A. L., & Gladwin, M. T. (2013) · Antioxidants & Redox Signaling · doi:10.1089/ars.2012.4833 · Cited by 68 · see citing works
Champlin, F. R., Ellison, M. L., Bullard, J. W., & Conrad, R. (2005) · International Journal of Antimicrobial Agents · doi:10.1016/j.ijantimicag.2005.04.020 · Cited by 66 · see citing works
Denitrifying Pseudomonas aeruginosa: some parameters of growth and active transport
Williams, D. R., Rowe, J. J., Romero, P. E., et al. (1978) · Applied and Environmental Microbiology · doi:10.1128/aem.36.2.257-263.1978 · Cited by 65 · see citing works
Catalase (KatA) Plays a Role in Protection against Anaerobic Nitric Oxide in Pseudomonas aeruginosa
Su, S., Panmanee, W., Wilson, J. J., Mahtani, H. K., Li, Q., VanderWielen, B. D., Makris, T. M., Rogers, M., McDaniel, C. T., Lipscomb, J. D., et al. (2014) · PLoS ONE · doi:10.1371/journal.pone.0091813 · Cited by 62 · see citing works
Shifting patterns of inhaled antibiotic use in cystic fibrosis
Moskowitz, S. M., Silva, S. J., Mayer-Hamblett, N., Pasta, D. J., Mink, D. R., Mabie, J. A., Konstan, M. W., & Wagener, J. S. (2008) · Pediatric Pulmonology · doi:10.1002/ppul.20873 · Cited by 61 · see citing works
Self-citationMajor, T. A., Panmanee, W., Mortensen, J. E., Gray, L. D., Hoglen, N., & Hassett, D. J. (2010) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.00379-10 · Cited by 61 · see citing works
A phase I clinical study of inhaled nitric oxide in healthy adults
Miller, C., Miller, M. K., McMullin, B., Regev-Shoshani, G., Serghides, L., Kain, K. C., ROAD, J. D., & Av‐Gay, Y. (2012) · Journal of Cystic Fibrosis · doi:10.1016/j.jcf.2012.01.003 · Cited by 60 · see citing works
Bourret, T. J., Boylan, J. A., Lawrence, K. A., & Gherardini, F. C. (2011) · Molecular Microbiology · doi:10.1111/j.1365-2958.2011.07691.x · Cited by 55 · see citing works
Bacterial host interactions in cystic fibrosis
Callaghan, M., & McClean, S. (2011) · Current Opinion in Microbiology · doi:10.1016/j.mib.2011.11.001 · Cited by 52 · see citing works
Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells
Moreau‐Marquis, S., Redelman, C. V., Stanton, B. A., et al. (2010) · Journal of Visualized Experiments · doi:10.3791/2186 · Cited by 45 · see citing works
Hon, Y. Y., Sun, H., Dejam, A., & Gladwin, M. T. (2010) · Drug Metabolism and Disposition · doi:10.1124/dmd.110.034355 · Cited by 27 · see citing works
Self-citationMcCaughey, G., Gilpin, D. F., Schneiders, T., Hoffman, L. R., McKevitt, M., Elborn, J. S., & Tunney, M. M. (2013) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.00750-13 · Cited by 24 · see citing works
Nitrate transport and its regulation by O2 in Pseudomonas aeruginosa
Hernandez, D., Dias, F. M., & Rowe, J. J. (1991) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(91)90022-b · Cited by 21 · see citing works
Mechanism of nitrite inhibition of cellular respiration inPseudomonas aeruginosa
Yang, T. (1985) · Current Microbiology · doi:10.1007/bf01567751 · Cited by 19 · see citing works
Acidified nitrite: a host defence against colonization with C. difficile spores?
Cunningham, R., Mustoe, E., Spiller, L. C., Lewis, S., & Benjamin, N. (2013) · Journal of Hospital Infection · doi:10.1016/j.jhin.2013.12.003 · Cited by 19 · see citing works
Reszka, K. J., Xiong, Y., Sallans, L., Pasula, R., Olakanmi, O., Hassett, D. J., & Britigan, B. E. (2012) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00172.2011 · Cited by 18 · see citing works
Wood, S. R., Firoved, A. M., Ornatowski, W., et al. (2006) · Free Radical Research · doi:10.1080/10715760601052610 · Cited by 15 · see citing works
Rasgado-Flores, H., Mandava, V. K., Siman, H., Driessche, W. V., Pilewski, J. M., Randell, S. H., & Bridges, R. J. (2013) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00166.2013 · Cited by 15 · see citing works
Self-citationQu, W., Zhou, Y., Sun, Y., et al. (2011) · The Journal of Microbiology · doi:10.1007/s12275-011-0262-7 · Cited by 12 · see citing works
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- Authors
- Zemke, A. C., Shiva, S., Burns, J. L., Moskowitz, S. M., Pilewski, J. M., Gladwin, M. T., & Bomberger, J. M.
- Affiliations
- University of Pittsburgh (US)
- University of Washington (US)
- Harvard University (US)
- Corresponding author
- Bomberger, Jennifer M.
- ORCID
- Venue
- Free Radical Biology and Medicine
- Publisher
- Elsevier BV
- Published
- 2014-12
- Volume
- 77
- Pages
- 307-316
- ISSN
- 0891-5849 (ISSN-L)
- PMC
- PMC4278422
- OpenAlex
- W2044668858
- Cited by
- 61
- References
- 54
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 4 · 2023: 8 · 2022: 4 · 2021: 8 · 2020: 5 · 2019: 3 · 2018: 4 · 2017: 3 · 2016: 12 · 2015: 4
- FWCI
- 2.73
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Free Radical Biology and Medicine (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- the Cystic Fibrosis Foundation
- Breathe Pennsylvania Research (2T32HL007563-26, B5P30DK089507-04, P30DK072506, R00HL098342-01)
- Cystic Fibrosis Foundation (R01HL098032, RO1HL096973, P01HL103455, T32 HL110849, MOSKOW13P0)
- the Institute for Transfusion Medicine and the Hemophilia Center of Western Pennsylvania (R01AI067653)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Cystic Fibrosis Research Advances
- Physical Sciences › Chemistry › Organic Chemistry › Antimicrobial agents and applications
- Keywords
- sodium nitrite, Pseudomonas aeruginosa, biofilm formation, antibiotic susceptibility, colistimethate, polymyxins, cystic fibrosis, airway epithelial cells, nitric oxide, nitrosative stress, nebulization, combination therapy, chronic airway infection
- MeSH terms
- Anti-Bacterial Agents, Bronchi, Cells, Cultured, Colistin, Epithelial Cells, Humans, Microbial Sensitivity Tests, Nitric Oxide, Oxygen, Pseudomonas aeruginosa, Sodium Nitrite, Biofilms, Drug Resistance, Bacterial
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