Role of Virulence Determinants in Candida albicans’ Resistance to Novel 2-bromo-2-chloro-2-(4-chlorophenylsulfonyl)-1-phenylethanone
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of 2-Bromo-2-Chloro-2-[(4-Chlorophenyl)Sulfonyl]-1-Phenylethanone (Compound 4)
2.2. Strains, Media, and Growth Conditions
2.3. Compound 4 Susceptibility Testing
2.4. Yeast Cell Growth Inhibition under Compound 4 in Time Intervals
2.5. Mononuclear Cell PBMCs. In Vitro Candida Stimulation
2.6. Fluorescence Microscopy
2.7. Cytokine Analysis
2.8. Assay of Candida Adherence to Human Line Caco-2 (ATCC HTB27, LGC, Poland). Inhibition of Adhesion: Microwell-Based Assay
2.9. Analysis of Gene Expression
2.10. Statistical Analysis
3. Results
3.1. In Vitro Anti-Candida Activity of Compound 4
3.2. Candida Cell Growth Inhibition under Compound 4
3.3. Compound 4 Treatment Results in Inhibition of Adhesive Properties of Candida Strains
3.4. Interaction of C. albicans Cells with Macrophages and Fungal Morphology under Compound 4
3.5. Compound 4 Modulates the KEX2 mRNA Expression
3.6. Interaction of Compound 4 with Interleukine 10
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Species | Strains | Parental | Genotype/Characteristic | Reference |
---|---|---|---|---|
Candida albicans | 90028 | None | Prototrophic wild-type strain | [12] |
SC5314 | None | Prototrophic wild-type strain | [13] | |
CAI4 | SC5314 | ura3Δ::1imm434/ura3Δ::1imm434 | [14] | |
cph1∆ | CAI4 | ura3Δ::1imm434/ura3Δ::1imm434 cph1Δ::hisGΔ/cph1::hisG-URA3-hisG | [15] | |
cph1∆ (CPH1) | CAI4 | ura3Δ::1imm434/ura3Δ::1imm434 cph1Δ::hisG/cph1Δ::hisG (CPH1) | [16] | |
efg1∆ | CAI4 | ura3Δ::1imm434/ura3Δ::1imm434 efg1Δ::hisG/efg1Δ::hisG-URA3-hisG | [16] | |
cph1∆/efg1∆ | CAI4 | ura3Δ::1imm434/ura3Δ::1imm434Δ cph1::hisG/cph1Δ::hisG Δefg1::hisG/efg1Δ::hisG-URA3-hisG | [16] | |
efg1∆ (EFG1) | CAI4 | ura3Δ::1 imm434/ura3Δ::1 imm434Δ efg1::hisG/efg1Δ::hisG (EFG1) | [16] | |
cph1∆/efg1∆ (EFG1) | CAI4 | ura3Δ::1 imm434/ura3Δ::1 imm434 cph1Δ::hisG/cph1Δ::hisG efg1Δ::hisG/efg1Δ::hisG (EFG1) | [16] | |
sap4∆ | SC5314 | sap4-1Δ::FRT/Δsap4-2::FRT | [17] | |
sap5∆ | SC5314 | sap5-1Δ::FRT/Δsap5-2::FRT | [17] | |
sap6∆ | SC5314 | sap6-1Δ::FRT/sap6-2 Δ::FRT | [17] | |
sap456∆ | SC5314 | sap4-1Δ::FRT/sap4-2Δ::FRT; Δsap5-1::FRT/sap5Δ-2::FRT; sap6-1Δ::FRT/sap6-2Δ::FRT | [17] | |
kex2∆CNA1 | CAI4 | ura3Δ::imm434/ura3Δ::imm434KEX2kex2Δ::hisGURA3hisG | [18] | |
kex2∆CNA2 | CAI4 | ura3Δ::imm434/ura3Δ::imm434KEX2kex2Δ::hisG | [18] | |
kex2∆CNA3 | CAI4 | ura3Δ::imm434/ura3Δ::imm434kex2Δ::hisGkex2Δ::hisGURA3hisG | [18] | |
kex2∆CNA4 | CAI4 | ura3Δ::imm434/ura3Δ::imm434kex2Δ::hisGkex2Δ::hisG | [18] | |
Candida glabrata | kex2∆1006 | Cg∆HT6 | his3Δ rp1Δt kex2Δ::HIS3 | [5] |
kex2∆1008 | 1005 | his3 Δtrp1Δ kex2Δ::HIS3 pKRT1(KEX2 TRP1) | [5] | |
kex2∆1010 | 1005 | his3Δ trp1Δ kex2Δ::HIS3 pACT-14(TRP1) | [5] |
Primers | Sequence (5′-3′) |
---|---|
ACT1-1 | GACAATTTCTCTTTCAGCACTAGTAGTGA |
ACT1-2 | GCTGGTAGAGACTTGACCAACCA |
KEX2-1 | TTTATATTGGGATATTTATTATCA |
KEX2-2 | TGGGATTTTAATAATAAAGGCAAA |
Candida Spp. | µg/mL | |||||||
---|---|---|---|---|---|---|---|---|
16 | 8 | 4 | 2 | 1 | 0.5 | 0.5 | 0.5 1 | |
SC5314 | 99.69 ± 0.01 | 99.55 ± 0.03 | 95.01 ± 0.01 | 92.70 ± 0.21 | 93.14 ± 0.05 | 95.59 ± 0.01 | 93.50 ± 0.01 | 100 |
Δsap4 3 | 97.97 ± 0.22 | 96.90 ± 0.02 | 95.50 ± 0.01 | 92.85 ± 0.02 | 100 ± 0.04 2 | 99.90 ± 0.03 | 98.89 ± 0.02 | 100 |
Δsap5 3 | 99.50 ± 0.00 | 99.31 ± 0.01 | 95.10 ± 0.20 | 40.05 ± 0.04 | 100 ± 0.03 2 | 98.32 ± 0.10 | 97.00 ± 0.04 | 100 |
Δsap6 3 | 99.80 ± 0.05 | 98.90 ± 0.01 | 99.42 ± 0.01 | 99.90 ± 0.01 | 96.95 ± 0.02 | 100 ± 0.01 2 | 97.67 ± 0.01 | 100 |
Δsap456 3 | 100 ± 0.02 | 98.70 ± 0.02 | 99.13 ± 0.01 | 99.85 ± 0.02 | 100 ± 0.01 | 97.89 ± 0.02 | 98.00 ± 0.04 | 100 |
Can16 3 | 99.40 ± 0.03 | 99.42 ± 0.04 | 99.94 ± 0.05 | 100 ± 0.216 | 100 ± 0.01 | 100 ± 0.04 2 | 99.00 ± 0.02 | 100 |
YLO323 | 99.71 ± 0.02 | 99.48 ± 0.03 | 95.67 ± 0.02 | 86.22 ± 0.02 | 92.04 ± 0.02 | 95.53 ± 0.22 | 95.67 ± 004 | 100 |
HLC52 | 99.63 ± 0.03 | 98.94 ± 0.04 | 99.48 ± 0.025 | 99.74 ± 0.06 | 99.77 ± 0.02 | 98.99 ± 0.04 | 98.90 ± 0.01 | 100 |
HLC54 3 | 99.75 ± 0.01 | 99.16 ± 0.09 | 99.33 ± 0.08 | 99.83 ± 0.05 | 100 ± 0.06 2 | 99.43 ± 0.09 | 99.00 ± 0.01 | 100 |
HLC74 3 | 99.76 ± 0.01 | 98.93 ± 0.04 | 98.96 ± 0.04 | 100 ± 0.02 | 99.38 ± 0.01 | 98.85 ± 0.03 | 98.50 ± 0.03 | 100 |
HLC84 | 99.71 ± 0.02 | 99.31 ± 0.01 | 99.24 ± 0.02 | 98.86 ± 0.04 | 99.15 ± 0.02 | 99.16 ± 0.03 | 99.00 ± 0.01 | 100 |
CNA1 | 99.06 ± 0.00 | 99.04 ± 0.00 | 99.00 ± 0.01 | 98.98 ± 0.01 | 98.97 ± 0.01 | 98.96 ± 0.01 | 98.00 ± 0.02 | 100 |
CNA2 | 99.05 ± 0.00 | 99.00 ± 0.01 | 98.99 ± 0.01 | 98.98 ± 0.00 | 98.970 ± 0.00 | 98.95 ± 0.00 | 98.77 ± 0.03 | 100 |
CNA3 | 99.44 ± 0.00 | 99.19 ± 0.00 | 99.17 ± 0.00 | 98.98 ± 0.00 | 98.77 ± 0.01 | 96.50 ± 0.00 | 96.00 ± 0.01 | 100 |
CNA4 | 99.96 ± 0.01 | 99.48 ± 0.00 | 99.12 ± 0.01 | 99.11 ± 0.01 | 98.67 ± 0.01 | 95.64 ± 0.01 | 94.78 ± 0.01 | 100 |
1006 | 100 ± 0.00 | 100 ± 0.21 2 | 99.47 ± 0.00 | 99.35 ± 0.01 | 99.27 ± 0.00 | 99.01 ± 0.01 | 99.00 ± 0.03 | 100 |
1008 | 99.08 ± 0.00 | 99.04 ± 0.01 | 98.99 ± 0.00 | 98.99 ± 0.01 | 98.97 ± 0.01 | 98.97 ± 0.00 | 97.90 ± 0.02 | 100 |
1010 | 100 ± 0.00 2 | 99.27 ± 0.01 | 99.21 ± 0.00 | 99.16 ± 0.00 | 98.94 ± 0.01 | 98.94 ± 0.00 | 98.94 ± 0.03 | 100 |
Species | Strains | Genotypes | Cells Treated at 0.25 µg/mL | Non Treated Cells |
---|---|---|---|---|
Candida albicans | SC5314 (wt) | Prototrophic wild-type strain | 29.99 ± 0.34 | 59.34 ± 0.06 |
sap4Δ | sap4-1Δ::FRT/Δsap4-2::FRT | 76.67 ± 0.15 | 46.32 ± 0.16 | |
sap5Δ | sap5-1Δ::FRT/Δsap5-2::FRT | 48.88 ± 0.17 | 35.55 ± 0.21 | |
sap6Δ | sap6-1Δ::FRT/sap6-2 Δ::FRT | 25.29 ± 0.16 | 22.99 ± 0.04 | |
sap4Δ/sap5Δ/sap6 | sap4-1Δ::FRT/sap4-2Δ::FRT; Δsap5-1::FRT/sap5Δ-2::FRT; sap6-1Δ::FRT/sap6-2Δ::FRT | 52.91 ± 0.33 | 15.88 ± 0.03 | |
cph1Δ | ura3Δ::1imm434/ura3Δ::1imm434 cph1Δ::hisG/cph1Δ::hisG | 63.89 ± 0.25 | 35.13 ± 0.24 | |
cph1Δ::CPH1 | ura3Δ::1imm434/ura3Δ::1imm434 cph1Δ::hisG/cph1Δ::hisG (CPH1) | 30.56 ± 0.03 | 12.82 ± 0.09 | |
efg1Δ | ura3Δ::1imm434/ura3Δ::1imm434 efg1Δ::hisG/efg1Δ::hisG-URA3-hisG | 46.40 ± 0.25 | 29.26 ± 0.16 | |
cph1Δ/efg1Δ | ura3Δ::1imm434/ura3Δ::1imm434Δ cph1::hisG/cph1Δ::hisG Δefg1::hisG/efg1Δ::hisG-URA3-hisG | 68.00 ± 0.16 | 72.21 ± 0.22 | |
efg1Δ::EFG1 | ura3Δ::1 imm434/ura3Δ::1 imm434Δ efg1::hisG/efg1Δ::hisG (EFG1) | 48.33 ± 0.12 | 8.34 ± 0.09 | |
cph1Δ/efg1Δ::EFG1 | ura3Δ::1 imm434/ura3Δ::1 imm434 cph1Δ::hisG/cph1Δ::hisG efg1Δ::hisG/efg1Δ::hisG (EFG1) | 61.87 ± 0.38 | 48.55 ± 0.078 | |
kex2Δ CNA1 | ura3Δ::imm434/ura3Δ::imm434KEX2kex2Δ::hisGURA3hisG | 37.85 ± 0.04 | 55.8 ± 0.18 | |
kex2Δ CNA2 | ura3Δ::imm434/ura3Δ::imm434KEX2kex2Δ::hisG | 22.49 ± 0.12 | 62.45 ± 0.27 | |
kex2Δ CNA3 | ura3Δ::imm434/ura3Δ::imm434kex2Δ::hisGkex2Δ::hisGURA3hisG | 48.17 ± 0.41 | 52.33 ± 0.16 | |
kex2Δ CNA4 | ura3Δ::imm434/ura3Δ::imm434kex2Δ::hisGkex2Δ::hisG | 7.33 ± 0.06 | 22.20 ± 0.06 | |
Candida glabrata | Cg kex2Δ 1006 | his3Δ rp1Δt kex2Δ::HIS3 | 45.13 ± 0.22 | 56.99 ± 0.24 |
Cg kex2Δ 1008 | his3Δtrp1Δ kex2Δ::HIS3 pKRT1 (KEX2 TRP1) | 17.43 ± 0.08 | 82.15 ± 0.19 | |
Cg kex2Δ 1010 | his3Δ trp1Δ kex2Δ::HIS3 pACT-14 (TRP1) | 18.34 ± 0.07 | 53.85 ± 0.28 |
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Staniszewska, M.; Bondaryk, M.; Ochal, Z. Role of Virulence Determinants in Candida albicans’ Resistance to Novel 2-bromo-2-chloro-2-(4-chlorophenylsulfonyl)-1-phenylethanone. J. Fungi 2017, 3, 32. https://doi.org/10.3390/jof3030032
Staniszewska M, Bondaryk M, Ochal Z. Role of Virulence Determinants in Candida albicans’ Resistance to Novel 2-bromo-2-chloro-2-(4-chlorophenylsulfonyl)-1-phenylethanone. Journal of Fungi. 2017; 3(3):32. https://doi.org/10.3390/jof3030032
Chicago/Turabian StyleStaniszewska, Monika, Małgorzata Bondaryk, and Zbigniew Ochal. 2017. "Role of Virulence Determinants in Candida albicans’ Resistance to Novel 2-bromo-2-chloro-2-(4-chlorophenylsulfonyl)-1-phenylethanone" Journal of Fungi 3, no. 3: 32. https://doi.org/10.3390/jof3030032