Anticytokine activity of Candida spp. and their ability to produce cytokine-like substances

O. A. Pashinina, Tatyana M. Pashkova, O. L. Kartashova, N. V. Morozova
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Abstract

Bacteria are known both to exhibit anticytokine activity (ACA), i.e., to secrete extracellular compounds which inactivate / neutralize various cytokines, and to produce cytokine-like substances (CPV) released to the culture medium. At the same time, the ability of Candida to secrete such substances has not been previously studied. The purpose of our study was to determine the presence of ACA and the ability to produce CPV in Candida strains isolated from the reproductive tract of apparently healthy pregnant women. The experimental series included 26 clinical strains of Candida spp. isolated from vaginal secretions. Isolation and identification of fungal species was based on morphological and biochemical criteria. ACA against IL-4, IL-6, IL-8, IL-10, IL-17A and TNF were detected following 2-hour co-incubation of fungal suspensions with solutions of the distinct cytokines at a ratio of 1:1. Determination of the cytokine concentrations was carried out by ELISA using the Cytokin kits (St. Petersburg). ACA amounts were defined as the percentage of cytokine inactivation in experimental samples compared to the control expressed as pg/mL. Their ability to produce CPV was expressed as percentage of cytokine production in experimental specimens compared to the controls (pg/mL). The obtained data were subjected to statistical evaluation. ACA of Candida spp. and their ability to produce CPV were revealed for the first time. with appropriate differences established between the studied Candida species, i.e., cultures of C. non-albicans showed ACA more often for the pro-inflammatory cytokines; C. albicans isolates showed more frequent production of the substances, similar to the pro-inflammatory cytokines IL-8, IL-17A, and TNF. Expression of ACA against IL-10 and ability to produce the anti-inflammatory IL-4- and IL-10-like substances were significantly higher in the cultures of C. non-albicans species. The results of these experiments expand our knowledge on the spectrum of Candida biological activities and require further study.
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念珠菌的抗细胞因子活性及其产生细胞因子样物质的能力
已知细菌既表现出抗细胞因子活性(ACA),即分泌胞外化合物,使各种细胞因子失活/中和,又产生细胞因子样物质(CPV)释放到培养基中。同时,念珠菌分泌这些物质的能力以前还没有被研究过。我们研究的目的是确定ACA的存在以及从表面健康的孕妇生殖道分离的念珠菌菌株产生CPV的能力。实验系列包括从阴道分泌物中分离的26株临床念珠菌。真菌种类的分离和鉴定是基于形态学和生化标准。将真菌悬浮液与不同细胞因子溶液以1:1的比例共孵育2小时,检测ACA对IL-4、IL-6、IL-8、IL-10、IL-17A和TNF的抑制作用。采用细胞因子试剂盒(Cytokin kit, St. Petersburg),采用ELISA法测定细胞因子浓度。ACA量定义为实验样品中细胞因子失活与对照组相比的百分比,以pg/mL表示。与对照组相比,它们产生CPV的能力以实验标本中细胞因子产量的百分比表示(pg/mL)。对所得数据进行统计评价。首次发现念珠菌的ACA及其产生CPV的能力。在所研究的念珠菌种类之间建立了适当的差异,即非白色念珠菌的培养更常显示促炎细胞因子的ACA;分离的白色念珠菌更频繁地产生这些物质,类似于促炎细胞因子IL-8、IL-17A和TNF。在非白色念珠菌中ACA抗IL-10的表达和产生抗炎IL-4和IL-10样物质的能力显著提高。这些实验结果扩大了我们对念珠菌生物活性谱的认识,需要进一步研究。
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