Article Figures & Data
Figures
- FIG. 1.
Infection of RHE by C. albicans. A total of 2 × 106C. albicans cells were used to infect the RHE, which was examined at 24 h by light microscopy. The strains used for infection were CAI4 (PMT/PMT), CAP1-3121(pRC18) (pmt1/pmt1), CAP1-3121(pCT30) (pmt1/pmt1[PMT1]), CAP4-2164(pRC18) (pmt4/pmt4), CAP4-2164(pSP1) (pmt4/pmt4[PMT4]), CAP2-2391(pRC18) (pmt6/pmt6), and CAP2-2391(pCT34) (pmt6/pmt6[PMT6]). Note that epithelial invasion caused by C. albicans CAI4 and PMT-reconstituted strains (dark spheres and filaments) is accompanied by vacuolization and edema of all keratinocyte layers.
- FIG.2.
Histological appearance of EHOM not infected or infected with C. albicans strains. An uninfected EHOM has an epidermis (Ep) containing an outmost stratum corneum layer (S.C.) and a basal layer (B. L.), as well as a collagen-containing dermis (D) that contains some fibroblasts (F). Cuboidal cells in the basal layer are indicated by asterisks, and large differentiated cells are indicated by white arrows. Following 22 h of infection, biopsies were taken from each EHOM, and Masson trichrome staining was performed. Sections of stained tissues were observed with an optical microscope and photographed. Representative photographs from three different experiments are shown (magnification, ×250). (A) Comparison of URA3-reconstituted pmt mutants with control strain CAF2-1. Mutants SPCa2 (pmt1), SPCa6 (pmt4), SPCa10 (pmt5), and SPCa8 (pmt6) were used for infection. (B) Comparison of PMT-reconstituted mutants and unreconstituted mutants. Strains CAP1-3121(pRC18) (pmt1/pmt1), CAP1-3121(pCT30) (pmt1/pmt1[PMT1]), P5-5744(pRC18) (pmt5/pmt5), and P5-5744(pBK1) (pmt5/pmt5) were compared.
- FIG. 3.
Virulence of C. albicans in an HDC model. CD2F1 mice were infected with C. albicans strains (1 × 106 cells/mouse). (A) Survival curves determined by the Kaplan-Meier method. Strains CAF2-1 (•), CAP4-2164(pSP1) (pmt4/pmt4 [PMT4]) (▪), CAP4-2164(pRC18) (pmt4/pmt4) (▴), P5-5744(pBK1) (pmt5/pmt5 [PMT5]) (▾), and P5-5744(pRC18) (pmt5/pmt5) (⧫) were tested. (B) Viable C. albicans cells in the kidneys and brain. Mice were treated with C. albicans strains (1 × 106 cells/mouse). At different times, mice were killed, and viable yeasts (CFU) in the kidneys (grey bars) and brain (black bars) were counted by plating samples of homogenized tissues on Sabouraud agar. The data are the means ± standard deviations for five mice.
Tables
- TABLE 1.
C. albicans strains
Strain Genotype Reference(s) CAF2-1 ura3Δ::imm434/URA3 10 CAI4 ura3Δ::imm434/ura3Δ::imm434 10 URA3-reconstituted strains SPCa2, SPCa3 As CAI4, but pmt1Δ::hisG/pmt1Δ::hisG ura3Δ::imm434/URA3 22 SPCa4, SPCa5 As CAI4, but pmt2Δ::hisG/PMT2 ura3Δ::imm434/URA3 22 SPCa6, SPCa7 As CAI4, but pmt4Δ::hisG/pmt4Δ::hisG ura3Δ::imm434/URA3 22 SPCa10, SPCa11 As CAI4, but pmt5Δ::hisG/pmt5Δ::hisG ura3Δ::imm434/URA3 22 SPCa7, SPCa8 As CAI4, but pmt6Δ::hisG/pmt6Δ::hisG ura3Δ::imm434/URA3 22 PMT-reconstituted strains and controls CAP1-3121(pCT30) As CAI4, but pmt1Δ::hisG/pmt1Δ::hisG (PMT1-URA3::LEU2) 32, this study CAP1-3121(pRC18) As CAI4, but pmt1Δ::hisG/pmt1Δ::hisG (URA3::LEU2) 32, this study CAP4-2164(pSP1) As CAI4, but pmt4Δ::hisG/pmt4Δ::hisG (PMT4-URA3::LEU2) 22 CAP4-2164(pRC18) As CAI4, but pmt4Δ::hisG/pmt4Δ::hisG (URA3::LEU2) 22 P5-5744(pBK1) As CAI4, but pmt5Δ::hisG/pmt5Δ::hisG (PMT5-URA3::LEU2) 22 P5-5744(pRC18) As CAI4, but pmt5Δ::hisG/pmt5Δ::hisG (URA3::LEU2) 22 CAP2-2391(pCT34) As CAI4, but pmt6Δ::hisG/pmt6Δ::hisG (PMT6-URA3::LEU2) 34, this study CAP2-2391(pRC18) As CAI4, but pmt6Δ::hisG/pmt6Δ::hisG (URA3::LEU2) 34, this study - TABLE 2.
Damage of reconstituted human epithelium by C. albicans strains
Expt Type of strains C. albicans strain Relevant genotype Amt of LDH released (U/liter)a A URA3 reconstituted Uninfected 10 ± 0 CAF2-1 Wild type 460 ± 10 SPCa2 pmt1/pmt1 90 ± 0b SPCa4 pmt2/PMT2 180 ± 10b SPCa6 pmt4/pmt4 190 ± 10b SPCa10 pmt5/pmt5 320 ± 10b SPCa8 pmt6/pmt6 140 ± 10b B PMT, reconstituted and controls Uninfected 20 ± 0 CAI4 Wild type 250 ± 10 CAP1-3121(pRC18) pmt1/pmt1 150 ± 10 CAP1-3121(pCT30) pmt1/pmt1 (PMT1) 210 ± 0b CAP4-2164(pRC18) pmt4/pmt4 80 ± 0 CAP4-2164(pSP1) pmt4/pmt4 (PMT4) 240 ± 10b P5-5744(pRC18) pmt5/pmt5 270 ± 0 P5-5744(pBK1) pmt5/pmt5 (PMT5) 270 ± 0 CAP2-2391(pRC18) pmt6/pmt6 110 ± 10 CAP2-2391(pCT34) pmt6/pmt6 (PMT6) 270 ± 10b - TABLE 3.
Virulence of pmt mutants in a mouse model of hematogenously disseminated infection
C. albicans straina Relevant genotype Mean survival time (days) No. of dead mice/total no.b SPCa2 pmt1/pmt1 >60c 0/6 SPCa3 pmt1/pmt1 >60c 0/6 SPCa4 pmt2/PMT2 >60c 0/6 SPCa5 pmt2/PMT2 >60c 0/6 SPCa6 pmt4/pmt4 3.5d 6/6 SPCa7 pmt4/pmt4 6.5c 6/6 SPCa10 pmt5/pmt5 4d 6/6 SPCa11 pmt5/pmt5 3.5d 6/6 SPCa8 pmt6/pmt6 3d 6/6 SPCa9 pmt6/pmt6 3d 6/6 CAF2-1 Wild type 2 6/6
