BACTERIA-BACTERIA AND HOST-BACTERIA INTERACTION OF BACTERIA ISOLATED FROM A PATIENT WITH CYSTIC FIBROSIS.

Gabriela Delgado, Juan Carlos Valdez, Nadia Gobbato, Silvia Orosco, Pablo Saguir, Clara Silva, Marcela Ortiz Mayor, Luis Herrera, Mirta Rachid, Nilda Noemi Arias, Ana Trejo

Abstract


Bacterial populations and inammation in the airways have been suggested as the main causes of cystic brosis (CF) pulmonary exacerbation. In
this work we studied the interactions of pathogenic bacteria Pseudomonas aeruginosa (Pae) and of Burkholderia cepacia complex (BCC) with the
oropharyngeal ora Streptococcus milleri group (SMG) isolated from the sputum of a CF patient with pulmonary exacerbation. We investigated the
virulence of single and mixed bacteria by evaluating the in vitro production of pyocyanin by spectrophotometric analysis and of rhamnolipids by
haemolysis and growth inhibition of Bacillus subtilis. Elastase production was analyzed in a mucus-like medium with the addition of
polymorphonuclear leukocytes (PMN) or DNA. Also, necrosis and netosis induced by bacteria in PMN were measured by ow cytometry and
spectrouorometry, respectively. Bacterial infections in a murine experimental model were analyzed.
Combinations of Pae with BCC and SMG enhanced pyocyanin and rhamnolipids production (p<0.01). Pae and BCC induced the highest elastase
release from PMN, but it was lower when SMG was added (p<0.05). Highest PMN necrosis levels were induced by the mixture of SMG, BCC, and
Pae (p<0.01). Pae was the major inducer of PMN netosis, followed by BCC and SMG. When Pae was combined with BCC and/or SMG, netosis
was reduced. Netosis correlated with elastase values.
In mouse models, infections combining SMG with Pae increase pulmonary inammation.
Commensal strains can increase the virulence of pathogenic bacteria. The exact role of Netosis and elastase in polymicrobial infections remains to
be determined.


Keywords


Cystic Fibrosis, Polymicrobial Infection, Streptococcus Milleri, Neutrophils, Netosis.

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References


Peters BM, Jabra-Rizk MA, O’May GA, Costerton W, Shirtliff ME. Polymicrobial Interactions: Impact on Pathogenesis and Human Disease.Clin Microbiol Rev 2012; 25: 193-213.

Gibson RL, Burns JL, Ramsey BW. Pathophysiology and management of pulmonary infections in cystic fibrosis. Am J Resp Crit Care Med 2003; 168: 918–51.

Alhede M, Bjarnsholt T, Jensen Pet al. Pseudomonas aeruginosa recognizes and responds aggressively to the presence of polymorphonuclear leukocytes. Microbiology 2009; 155: 3500–58.

Smith JJ, Travis SM, Greenberg EP, Welsh MJ. Cystic fibrosis airway epithelia fail to kill bacteria because of abnormal airway surface fluid. Cell 1996; 85:229-36.

Govan JR, Deretic V. Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev 1996; 60: 539-74.

Rahman S and Gadjeva M. Does NETosis contribute to the bacterial pathoadaptation in cystic fibrosis? Front Immunol 2014; 5:378. doi: 10.3389/fimmu.2014.00378

LiPuma JJ. The changing microbial epidemiology in cystic fibrosis. Clin Microbiol Rev 2010; 23: 299-323.

Downey DG, BellSC, Elborn JS. Neutrophils in cystic fibrosis. Thorax2009; 64: 81–8.

Harrison F. Microbiology ecology of the cystic fibrosis lung. Microbiology 2007; 153: 917-23.

Sibley CD, Parkins MD, Rabin HR, Duan K, Norgaard JC, Surette MG. A polymicrobial perspective of pulmonary infections exposes an enigmatic pathogen in cystic fibrosis patients. Proc Natl Acad Sci USA 2008; 105: 15070-75.

Duan K, Dammel C, Stein J, Rabin H, Surette MG. Modulation of Pseudomonas aeruginosa gene expression by host microflora through interspecies communication. Mol Microbiol 2003; 50: 1477-91.

Burns JL, Emerson J, Stapp JR, et al. Microbiology of sputum from patients at cystic fibrosis centers in the United States. Clin Infect Dis 1998; 27: 158–63.

Johnson Mk, Boese-Marrazzo D. Production and properties of heat-stable extracellular hemolysin from Pseudomonas aeruginosa. Infect Immun 1980; 29: 1028-33.

Koch AK, Käpelli O, Fiechter A, Reiser J. Hydrocarbon Assimilation and Biosurfactant Production in Pseudomonas aeruginosa Mutants. J Bacteriol 1991; 173: 4212-19.

Yeung ATY, Parayno A, Hancock REW.Mucin Promotes Rapid Surface Motility in Pseudomonas aeruginosa. mBio 2012; mBio 3(3):e00073-12. doi:10.1128/mBio.00073-12.

Cheng OZ and Palaniyar N. NET balancing: a problem in inflammatory lung diseases. Front Immun 2013; 4:1. doi: 10.3389/fimmu.2013.00001

Gambello MJ, Iglewski BH. Cloning and characterization of the Pseudomonas aeruginosa lasR gene, a transcriptional activator of elastase expression. J Bacteriol 1991; 173: 3000–9.

Ramos AN, Gobbato N, Rachid M, González L, Yantorno O, Valdez JC. Effect of Lactobacillus plantarum and Pseudomonas aeruginosa culture supernatants on polymorphonuclear damage and inflammatory response. Int Immunopharmacol 2010; 10: 247-51

Gray RD, Lucas CD, MacKellar A et al. Activation of conventional protein kinase C (PKC) is critical in the generation of human neutrophils extracellular traps. J Inflamm 2013; 10: 12-20.

Hirche TO, Atkinson JJ, Bahr S, Belaaouaj A. Deficiency in neutrophil elastase does not impair neutrophil recruitment to inflamed sites. Am J Respir Cell Mol Biol 2004; 30: 576-84.

Allen L, Dockrell DH, Pattery T et al. Pyocyanin Production by Pseudomonas aeruginosa Induces Neutrophil Apoptosis and Impairs Neutrophil-Mediated Host Defenses In Vivo. J Immunol2005; 174: 3643-9.

Klepac-Ceraj V, Lemon KP, Martin TR, et al. Relationship between cystic fibrosis respiratory tract bacterial communities and age, genotype, antibiotics and Pseudomonas aeruginosa. Environ Microbiol 2010; 12: 1293–303.

Sibley CD, Sibley KA, Leong TA et al. The Streptococcus milleri population of a cystic fibrosis clinic reveals patient specificity and intraspecies diversity. J Clin Microbiol 2010; 48: 2592-4.

Wahab AA, JanahiIA, Marafia MM; El-Shafie S. Microbiological identification in cystic fibrosis patients with CFTR 1234V mutation. J Trop Pediatr 2004; 50: 229–33.

Moore JE, Shaw A, Millar BC, Downey DG, Murphy PG; Elborn JS. Microbial ecology of the cystic fibrosis lung: does microflora type influence microbial loading?. Br J Biomed Sci 2005; 62: 175–8.

O’Sullivan BP, Freedman SD. Cystic fibrosis. Lancet 2009;373: 1891–904.

Chmiel J, Berger M, Konstan M. The role of inflammation in the pathophysiology of CF lung disease. Clin Rev Allergy Immunol2002; 23: 5–27.

Yoo D, Winn M, Pang L, Moskowitz SM, et al. Release of Cystic Fibrosis AirwayInflammatory Markers from Pseudomonas aeruginosa–Stimulated Human Neutrophils Involves NADPH Oxidase-Dependent Extracellular DNA Trap Formation. J Immunol 2014; 192: 4728–38.

Shah P, Scott S, Knight R, Hudson M. The effects of recombinant human DNase on neutrophil elastase activity and interleukin-8 levels in the sputum of patients with cystic fibrosis. Eur Respir J1996; 9: 531–4.


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