Analysis of the cytotoxic properties of collection strains of coccidioidomycosis causative agents

D. Tereshko, I. Novitskaya
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Abstract

Introduction. C. immitis and C. posadasii are primary pathogens and are of reasonable interest in terms of their biological properties and, in particular, cytotoxic characteristics. Aim. To study the cytotoxicity of strains of coccidioidomycosis causative agents using the MTT test on in vitro models of cell lines. Materials and methods. Strains Coccidioides spp. were cultivated in a nutrient medium intended for cell lines. Cultures Wehi 3 and HEp-2 were exposed to sterile filtrates of the growth medium of causative agents of coccidioidomycosis in various dilutions, followed by measuring the mitochondrial activity of cells in the MTT-assay on day 4. The cytotoxic properties of fungi of the genus Coccidioides were assessed by calculating the cytotoxic concentration (СС50) of the metabolic products of micromycetes. Results. The most adequate cell model was selected, as well as the conditions for setting the MTT test for the in vitro study of the cytopathogenic effects of micromycetes of the genus Coccidioides were worked out. It was found that the strains Coccidioides spp. have different intensities of metabolite production, which, in turn, does not directly correlate with their cytotoxicity. By calculating the cytotoxic concentration (CC50), data were obtained on the cytotoxic properties of 7 collection strains of coccidioidomycosis pathogens in the mycelial growth phase. Conclusion. Based on the results of assessing the mitochondrial activity of cells of the lines Wehi 3 and HEp-2, the pronounced cytotoxic properties of the collection strains of coccidioidomycosis causative agents, which were subcultured in vitro for a long time, were confirmed.
球虫病病原收集菌株的细胞毒性分析
介绍。C. immitis和C. posadasii是原发病原体,就其生物学特性,特别是细胞毒性特性而言,它们具有合理的兴趣。的目标。目的:用MTT法研究球虫菌病病原菌的细胞毒性。材料和方法。在用于细胞系的营养培养基中培养球虫。将Wehi 3和HEp-2培养物以不同稀释度暴露于球虫菌病病原体生长培养基的无菌滤液中,第4天用mtt法测定细胞线粒体活性。通过计算微菌代谢产物的细胞毒浓度(СС50)来评估球虫属真菌的细胞毒特性。结果。选择了最合适的细胞模型,并确定了设置MTT试验体外研究球虫属微菌的细胞致病作用的条件。研究发现,球虫具有不同强度的代谢物产生,而这与它们的细胞毒性没有直接关系。通过计算细胞毒浓度(CC50),获得7株球孢子菌病病原菌菌丝生长期的细胞毒特性数据。结论。通过对微粉3号和HEp-2细胞系线粒体活性的测定,证实了长时间体外传代培养的球孢子菌病原菌收集株具有明显的细胞毒性。
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