MGHPGA

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 Womens
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
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
Plt.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