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

4. Percentage probability
Summary of Results from Seven CF Mouse Colonization Studies Probability of Colonization Negatively Correlates with CFTR Expression
100
P<.001 75
P=.02
Percentage probability
of colonization 50 25
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 80
PA14 Wild Type
PA14 algD 60
PA14 gacA 40 20 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 1 3 7 11 15 17 19 21 23
Weeks after Infection with P. aeruginosa algD

7. Weeks After Infection with P. aeruginosa gacA
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 1 3 5 7 11 15 19 21 23 25 27
Weeks After Infection with P. aeruginosa gacA