研究嵌合型干扰素α-2b 对动植物真菌病原体的杀菌作用

V. S. Maslennikova, C. M. Miroshnichenko, E. V. Shelikhova, K. A. Tabanyukhov, I. Y. Deulin, M. B. Pykhtina
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摘要

本研究的目的是评估重组干扰素α-2b与人类脂蛋白A-I融合的嵌合形式(在皮奇亚酵母中生物合成)对重要真菌病原体(人类和植物病害的病原体)的杀菌效果。采用琼脂块法评估了嵌合细胞因子对真菌病原体的杀菌活性:用亚甲蓝染色的真菌菌体切片进行肉眼观察和显微镜观察评估杀菌效果,用亚培养法评估真菌孢子的萌发,用 Vero 细胞培养法评估干扰素制剂的细胞毒性。所研究的重组干扰素能抑制病原真菌的生长和繁殖。当浓度为 15 ng/ml 时,嵌合 IFN 对病原真菌 A.交替菌和青霉的作用最大;当浓度为 150 ng/ml 时,对真菌 F. oxysporum 和曲霉孢子的抑制率高达 87%。新嵌合细胞因子的细胞毒性为 5.8 μg/ml,明显高于真菌的毒性浓度。嵌合型干扰素α-2b可用作具有杀真菌活性的免疫治疗剂。
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Study of the fungicidal effect of the chimeric form of interferon alpha-2b in relation to fungi-pathogens of plants and animals
The purpose of this study is to evaluate the fungicidal effect of a chimeric form of recombinant interferon alpha-2b, fused with human apolipoprotein A-I, obtained by biosynthesis in the yeast Pichia рastoris, against significant fungal pathogens – pathogens of human and plant diseases.Material and methods. The fungicidal activity of the chimeric cytokine was assessed using the agar block method against fungal pathogens: Alternaria alternata, Fusarium oxysporum, Aspergillus spp. and Penicillum spp. The fungicidal effect was assessed visually and by microscopy of a section of the fungal body stained with methylene blue, the germination of fungal spores – by subculture, the cytotoxicity of interferon preparations – on Vero cell culture.Results and discussion. The studied recombinant interferons suppress the growth and reproduction of pathogenic fungi. Chimeric IFN has the greatest effect against pathogenic fungi A. alternate, Penicillum at a concentration of 15 ng/ml; inhibition of sporulation of fungi F. oxysporum and Aspergillus up to 87 % is achieved at a concentration of 150 ng/ml. The cytotoxicity of the new chimeric cytokine is 5,8 μg/ml, which is significantly more than toxic concentrations for fungi.Conclusions. The chimeric form of interferon alpha-2b can be used as an immunotherapeutic agent with fungicidal activity.
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