Peptide ZP2, a synthetic analogue of the active center of granulocyte-macrophage colony stimulating factor (GM-CSF), exhibits a wide range of immunobiological effects, including antibacterial activity. At the same time, its effect (in sub-inhibitory concentrations) on the biological properties of Enterococcus spp., the causative agents of many infectious and inflammatory diseases remains poorly understood. The aim of our study was to analyze the nature of the effect of synthetic peptide ZP2 on anticytokine activity (ACA) and its ability to produce cytokine-like substances (CLS) in Enterococcus spp. 18 clinical isolates of Enterococcus spp. were used. Over the experiments, the bacterial strains were cultured in Schaedlers broth with ZP2 peptide at 37 C for 24 hours. No specific peptide was added in the control. ACA for IL-8, TNF and IL-17A and the production of the corresponding CLS were determined by the ELISA method. To assess ACA, the proportion of cytokine inactivation in the experiment relative to the control was calculated and expressed in pg/ml; CLS production was evaluated by the level of cytokines in the experiment and control, expressed as pg/ml. The data were subjected to statistical processing. It was revealed that Enterococcus spp. strains are capable of secreting compounds that inactivate cytokines IL-8, IL-17A and TNF, and produce CLS in the culture medium. Intragenital and intraspecific variability was noted in the presence and frequency of occurrence and in the severity of these properties. It was found that the ZP2 peptide in E. faecium increases ACA with respect to all studied cytokines. When tested with E. faecalis, it either did not affect their ACA against TNF and IL-8, or completely inhibited ACA for IL-17A. At the same time, ZP2 blocked the production of CLS, e.g., IL-17A and IL-8 in E. faecium, but increased the production of CLS similar to TNF, and, with E. faecalis, it increased the number of IL-17A-producing isolates twofold, although the average level of production of these CLS was lower than in the control. Enterococcus spp strains are capable of secreting compounds that inactivate cytokines IL-8, TNF and IL- 17A, and may produce substances similar to these cytokines. The synthetic peptide ZP2 has a modifying effect on the manifestation of these properties by Enterococci. Further studies of biological and pathogenetic features of Enterococci and other bacterial species, as well as modifying effects of the ZP2 peptide are required.

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.

Objective. To characterize pathogenic pheno- and genoprofiles of staphylococci of different species, isolated from the secretion of the prostate gland in male patients with chronic bacterial prostatitis.Materials and methods. The bacterial spectrum of microflora was studied by a bacteriological method; the species identification of microorganisms was carried out by mass spectrometry. Detection of genes determining pathogenicity factors was carried out by PCR. The biofilm-forming ability of staphylococci, as well as their anti-lysozyme, hemolytic, and adhesive activity, were determined by photometry.Results. Staphylococci were found to be dominant in the pathogen structure of chronic bacterial prostatitis. These microorganisms, regardless of their species, had a pronounced pathogenic potential. Specific features in the bioprofiles of cultures of different species were determined according to the severity of the studied biological properties. Thus, the hemolytic activity and biofilm-forming ability was significantly higher in S. aureus. Conversely, CNS strains were characterized by high anti-lysozyme activity. A significant difference was noted in the prevalence of genes that determine pathogenicity factors in the studied staphylococci of different species.Conclusion. The pathogenic bioprofile of staphylococci of various species isolated from patients with chronic bacterial prostatitis can be used as a criterion in the search and identification of the pathogen, as well as in the development of effective therapeutical approaches.

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The synthetic analogue of the active center of GM-CSF — peptide ZP2 has a wide range of biological effects. However, its effect on the biological properties of Candida has not been previously investigated. The aim is to analyze the nature of the effect of synthetic peptide ZP2 on anti–cytokine activity (ACA) and the ability to produce cytokine-like substances (CLS) in Candida. Materials and methods. 26 vaginal strains of Candida spp. were used in the experiment. from conditionally healthy pregnant women. To study the effect of the ZP2 peptide on the ACA of Candida and their CLS production, Candida were cultured in Saburo broth with the ZP2 peptide at 37oC for 24 hours. The ZP2 peptide was not added to the control. ACA for IL-8, TNFα and IL17A and CLS products were determined by the ELISA method according to a well-known technique in the original modification using kits of Cytokin LLC (St. Petersburg). To assess the ACA of Candida, the proportion of cytokine inactivation in the experiment relative to the control was calculated and expressed in pg/ml; the production of CLS by Candida was judged by the level of cytokines in the experiment and control, expressed in pg/ml. The data were subjected to statistical processing. Results and discussion. The multidirectional effect of the peptide ZP2 on the ACA of Candida with respect to a number of cytokines and their ability to produce individual CLS has been established. The ZP2 peptide did not change the incidence of ACA against IL8 in Candida spp. strains, but increased the severity of this trait in C. non-albicans isolates. The ZP2 peptide inhibited ACA against TNFα in C. albicans, but stimulated the manifestation of ACA against IL17A only in C. non-albicans. The ZP2 peptide slightly increased the proportion of C. albicans with the ability to produce substances similar to IL8 and TNFα without significantly changing the severity of the trait. Among C. non-albicans, the number of strains producing these substances did not change significantly under the action of the ZP2 peptide, but the expression of the trait increased. The ZP2 peptide reduced the proportion of Candida spp. strains capable of producing IL17A-like substances. Conclusion. Candida are able to secrete compounds that inactivate various cytokines, as well as produce cytokine-like substances. The synthetic analogue of the active center of GM-CSF-peptide ZP2 has a modifying effect on the manifestation of these properties by fungi.