Abstract
Cryptococcus gattii isolates from the Pacific Northwest have exhibited higher fluconazole MICs than isolates from other sites. The mechanism of fluconazole resistance in C. gattii is unknown. We sought to determine the role of the efflux pumps Mdr1 and Pdr11 in fluconazole susceptibility. Using biolistic transformation of the parent isolate, we created a strain lacking Mdr1 (mdr1 Δ) and another strain lacking Pdr11 (pdr11 Δ). Phenotypic virulence factors were assessed by standard methods (capsule size, melanin production, growth at 30 and 37°C). Survival was assessed in an intranasal murine model of cryptococcosis. Antifungal MICs were determined by the M27-A3 methodology. No differences in key virulence phenotypic components were identified. Fluconazole susceptibility was unchanged in the Mdr1 knockout or reconstituted isolates. However, fluconazole MICs decreased from 32 μg/ml for the wild-type isolate to < 0.03 μg/ml for the pdr11Δ strain and reverted to 32 μg/ml for the reconstituted strain. In murine models, no difference in virulence was observed between wild-type, knockout, or reconstituted isolates. We conclude that Pdr11 plays an essential role in fluconazole susceptibility in C. gattii. Genomic and expression differences between resistant and susceptible C. gattii clinical isolates should be assessed further in order to identify other potential mechanisms of resistance.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1202-1207 |
| Number of pages | 6 |
| Journal | Antimicrobial Agents and Chemotherapy |
| Volume | 60 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 1 2016 |
Fingerprint
ASJC Scopus subject areas
- Pharmacology
- Infectious Diseases
- Pharmacology (medical)
Cite this
Fluconazole susceptibility in Cryptococcus gattii is dependent on the ABC transporter Pdr11. / Yang, Mai Lee; Uhrig, John; Vu, Kiem; Singapuri, Anil; Dennis, Michael; Gelli, Angela C; Thompson, George Richard.
In: Antimicrobial Agents and Chemotherapy, Vol. 60, No. 3, 01.03.2016, p. 1202-1207.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Fluconazole susceptibility in Cryptococcus gattii is dependent on the ABC transporter Pdr11
AU - Yang, Mai Lee
AU - Uhrig, John
AU - Vu, Kiem
AU - Singapuri, Anil
AU - Dennis, Michael
AU - Gelli, Angela C
AU - Thompson, George Richard
PY - 2016/3/1
Y1 - 2016/3/1
N2 - Cryptococcus gattii isolates from the Pacific Northwest have exhibited higher fluconazole MICs than isolates from other sites. The mechanism of fluconazole resistance in C. gattii is unknown. We sought to determine the role of the efflux pumps Mdr1 and Pdr11 in fluconazole susceptibility. Using biolistic transformation of the parent isolate, we created a strain lacking Mdr1 (mdr1 Δ) and another strain lacking Pdr11 (pdr11 Δ). Phenotypic virulence factors were assessed by standard methods (capsule size, melanin production, growth at 30 and 37°C). Survival was assessed in an intranasal murine model of cryptococcosis. Antifungal MICs were determined by the M27-A3 methodology. No differences in key virulence phenotypic components were identified. Fluconazole susceptibility was unchanged in the Mdr1 knockout or reconstituted isolates. However, fluconazole MICs decreased from 32 μg/ml for the wild-type isolate to < 0.03 μg/ml for the pdr11Δ strain and reverted to 32 μg/ml for the reconstituted strain. In murine models, no difference in virulence was observed between wild-type, knockout, or reconstituted isolates. We conclude that Pdr11 plays an essential role in fluconazole susceptibility in C. gattii. Genomic and expression differences between resistant and susceptible C. gattii clinical isolates should be assessed further in order to identify other potential mechanisms of resistance.
AB - Cryptococcus gattii isolates from the Pacific Northwest have exhibited higher fluconazole MICs than isolates from other sites. The mechanism of fluconazole resistance in C. gattii is unknown. We sought to determine the role of the efflux pumps Mdr1 and Pdr11 in fluconazole susceptibility. Using biolistic transformation of the parent isolate, we created a strain lacking Mdr1 (mdr1 Δ) and another strain lacking Pdr11 (pdr11 Δ). Phenotypic virulence factors were assessed by standard methods (capsule size, melanin production, growth at 30 and 37°C). Survival was assessed in an intranasal murine model of cryptococcosis. Antifungal MICs were determined by the M27-A3 methodology. No differences in key virulence phenotypic components were identified. Fluconazole susceptibility was unchanged in the Mdr1 knockout or reconstituted isolates. However, fluconazole MICs decreased from 32 μg/ml for the wild-type isolate to < 0.03 μg/ml for the pdr11Δ strain and reverted to 32 μg/ml for the reconstituted strain. In murine models, no difference in virulence was observed between wild-type, knockout, or reconstituted isolates. We conclude that Pdr11 plays an essential role in fluconazole susceptibility in C. gattii. Genomic and expression differences between resistant and susceptible C. gattii clinical isolates should be assessed further in order to identify other potential mechanisms of resistance.
UR - http://www.scopus.com/inward/record.url?scp=84960155687&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84960155687&partnerID=8YFLogxK
U2 - 10.1128/AAC.01777-15
DO - 10.1128/AAC.01777-15
M3 - Article
C2 - 26643330
AN - SCOPUS:84960155687
VL - 60
SP - 1202
EP - 1207
JO - Antimicrobial Agents and Chemotherapy
JF - Antimicrobial Agents and Chemotherapy
SN - 0066-4804
IS - 3
ER -