摘要
Background Candida albicans （C. albicans） strains can spontaneously switch at a very low frequency from white to opaque phase. The ability to switch reversibly between white and opaque phenotype and contributes to the pathogenicity of C. albicans. White and opaque switching can be induced by various environmental signals. Previous study showed that opaque cells switch en masse to white when transferred in vitro to 37℃, the temperature of their animal host. The objective of the present study was to determine the effect of different concentration of carbon dioxide and temperature on white-opaque switching, and to determine the different anti-candida killing activity of white and opaque form by human monocyte-macrophage cell line THP-1.Methods White-opaque switching and opaque-white switching were assayed. Modified Lee＇s medium supplemented with phloxine B was used to detect white and opaque forms of C. albicans under 0.03% CO2 at 25℃, 0.03% CO2 at 37℃ and 5% CO2 at 37℃. Growth curve of C. albicans was monitored using OD value at 630 nm simultaneously. White and opaque forms of C. albicans and THP-1 cells were cocultured at ratio of 1：10. Colony serial dilutions were used to assay for intracellular candidacidal activity. MTT assay was used to measure the extracellular candidacidal activity.Results Phenotype switching was successfully induced in vitro in all three strains of C. albicans. When evaluating white to opaque switching, opaque colony proportion of all colonies was 0.572±0.087, 0.920±0.030 and 0.985±0.026 exposure of white cells to 0.03% CO2 at 25℃, 0.03% CO2 at 37℃ and 5% CO2 at 37℃. When evaluating opaque to white switching, opaque colony proportion of all colonies was 0.600±0.114, 0.983±0.003 and 0.998±0.003 exposure of white cells to 0.03% CO2 at 25℃, 0.03% CO2 at 37℃ and 5% CO2 at 37℃. No significant difference of white or opaque form growth rate was found among three conditions （P〉0.05）. THP-1 mediated extracellular anti-candida activity in white form was （79.80±3.71）% and （56.28±19.12）% at different dilution ratio, which were significantly lower than that in opaque form （100%, P 〈0.01）. THP-1 mediated intracellular anti-candida activity in white form （（62.98±5.02）%） was significantly lower than that in opaque form （（87.07±1.80）%, P 〈0.01）.Conclusions Our results showed that opaque form is more vulnerable and less virulent than that in white form. It suggested that higher concentration of CO2 and 37℃ in host niches stabilize the less virulent opaque cell of C. albicans,which might have implications for pathogenesis, commensalism and mating.
Background Candida albicans （C. albicans） strains can spontaneously switch at a very low frequency from white to opaque phase. The ability to switch reversibly between white and opaque phenotype and contributes to the pathogenicity of C. albicans. White and opaque switching can be induced by various environmental signals. Previous study showed that opaque cells switch en masse to white when transferred in vitro to 37℃, the temperature of their animal host. The objective of the present study was to determine the effect of different concentration of carbon dioxide and temperature on white-opaque switching, and to determine the different anti-candida killing activity of white and opaque form by human monocyte-macrophage cell line THP-1.Methods White-opaque switching and opaque-white switching were assayed. Modified Lee＇s medium supplemented with phloxine B was used to detect white and opaque forms of C. albicans under 0.03% CO2 at 25℃, 0.03% CO2 at 37℃ and 5% CO2 at 37℃. Growth curve of C. albicans was monitored using OD value at 630 nm simultaneously. White and opaque forms of C. albicans and THP-1 cells were cocultured at ratio of 1：10. Colony serial dilutions were used to assay for intracellular candidacidal activity. MTT assay was used to measure the extracellular candidacidal activity.Results Phenotype switching was successfully induced in vitro in all three strains of C. albicans. When evaluating white to opaque switching, opaque colony proportion of all colonies was 0.572±0.087, 0.920±0.030 and 0.985±0.026 exposure of white cells to 0.03% CO2 at 25℃, 0.03% CO2 at 37℃ and 5% CO2 at 37℃. When evaluating opaque to white switching, opaque colony proportion of all colonies was 0.600±0.114, 0.983±0.003 and 0.998±0.003 exposure of white cells to 0.03% CO2 at 25℃, 0.03% CO2 at 37℃ and 5% CO2 at 37℃. No significant difference of white or opaque form growth rate was found among three conditions （P〉0.05）. THP-1 mediated extracellular anti-candida activity in white form was （79.80±3.71）% and （56.28±19.12）% at different dilution ratio, which were significantly lower than that in opaque form （100%, P 〈0.01）. THP-1 mediated intracellular anti-candida activity in white form （（62.98±5.02）%） was significantly lower than that in opaque form （（87.07±1.80）%, P 〈0.01）.Conclusions Our results showed that opaque form is more vulnerable and less virulent than that in white form. It suggested that higher concentration of CO2 and 37℃ in host niches stabilize the less virulent opaque cell of C. albicans,which might have implications for pathogenesis, commensalism and mating.
基金
This work was partly supported by a grant from National Natural Science Foundation of China （No. 30671893）.
