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MGH-PGA Genomic Analysis of Stress and Inflammation:

MGH-PGA Genomic Analysis of Stress and Inflammation: Pseudomonas aeruginosa Infections of CF Mouse Models. Nicole T. Liberati, Dan G. Lee, Jacinto M. Villanueva and Frederick M. Ausubel Department of Molecular Biology Massachusetts General Hospital.

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MGH-PGA Genomic Analysis of Stress and Inflammation:

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  1. MGH-PGA Genomic Analysis of Stress and Inflammation: Pseudomonas aeruginosa Infections of CF Mouse Models Nicole T. Liberati, Dan G. Lee, Jacinto M. Villanueva and Frederick M. Ausubel Department of Molecular Biology Massachusetts General Hospital Carolyn Cannon, Fadie Coleman, Mike Kowalski Jeff Lyczak, Martin Lee, Gloria Meluleni, and Gerald Pier Channing Laboratory Brigham and Women’s Hospital

  2. Use of Transgenic CF Mice for Establishing Chronic Oropharyngeal Colonization • Difficult to establish chronic P. aeruginosa infections in CF KO mice. • Maintain mice 5-7 days on water containing antibiotics. • 107 CFU P. aeruginosa/ml in drinking water for 5 days. • Sterile water for 1 week followed by water containing antibiotics. • Monitor mice by throat culture every 1-2 weeks.

  3. SPC-CFTR* *Over-express CFTR via lung-specific surfactant protein C promoter P. aeruginosa PA14 Chronically Colonizes Oropharynx of CF Mice 100 80 Percent Positive Throat Cultures 60 C57sBl/6 WT mice CF mice 40 20 0 1 3 6 8 10 12 15 17 19 21 23 26 29 Weeks After Infection with P. aeruginosa PA14

  4. Summary of Results from Seven CF Mouse Colonization StudiesProbability of Colonization Negatively Correlates with CFTR Expression 100 P<.001 75 P=.02 Percentage probability of colonization 50 25 0 CF mice Wild-type mice SPC-CFTR mice Mouse strain

  5. PA14 algD (alginate) and gacA (global accessory regulator) Mutants Fail to Colonize CF Mice 100 Percent Positive Throat Cultures PA14 Wild Type 80 PA14 algD 60 PA14 gacA 40 20 0 1 3 7 11 15 17 19 21 23 Weeks after Infection with P. aeruginosa

  6. PA14 algD (alginate) Mutant Fails to Chronically Colonize the Oropharynx of CF Mice PA14 algD in transgenic CF mice 100 Percent Positive Throat Cultures 80 C57Bl/6 WT-mice 60 CF mice 40 20 0 1 3 7 11 15 17 19 21 23 Weeks after Infection with P. aeruginosa algD

  7. PA14 gacA (global accessory regulator) Mutant Fails to Chronically Colonize the Oropharynx of CF Mice PA14 gacA in transgenic CF mice 100 Percent Positive Throat Cultures 80 C57Bl/6 WT-mice CF mice 60 40 20 0 1 3 5 7 11 15 19 21 23 25 27 Weeks After Infection with P. aeruginosa gacA

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