Changes in the Level of Cytokines of Human Peripheral Blood In Vitro under the Influence of Metabolites of Bacteria Isolated from Permafrost

About the authors:

Svetlana S. Kolyvanova, Junior Researcher, Department of Bioresources Сryosphere, Tyumen Scientific Centre of the Siberian Branch of the Russian Academy of Sciences; Рostgraduate Student, Department of Anatomy and Рhysiology of Нumans and Аnimals, University of Tyumen; kolyvanova93@mail.ru

Lyudmila F. Kalenova, Dr. Sci. (Biol.), Chief Researcher, Department of Bioresources Сryosphere, Tyumen Scientific Centre of the Siberian Branch of the Russian Academy of Sciences; Leading Researcher, International Center of Cryology and Cryosophy, University of Tyumen; lkalenova@mail.ru

Abstract:

Microbiological research has shown viable microorganisms in the permafrost. The survival of bacteria in extremely cold conditions was the basis for the study of their byproduct (secondary metabolites). It is known that the bacterial cells’ number and their cultivation their cultivation temperature significantly affect their biological activity. For example, an experimental study of M3 strain of the Bacillus sp., isolated from permafrost, showed that the changes in cultivation temperature affected the enzymatic, immunotropic, and reparative properties of bacteria in vitro and in vivo. In this regard, the study of the effect of metabolites of microorganisms at the level of cytokine secretion, as indicators of immunological reactivity, seems relevant to create new immunotropic drugs in the future.

The effect of secondary metabolites of the bacteria Bacillus sp. (strain M3) and Bacillus megaterium (strain 8/75-1), isolated from permafrost of different geological ages, on the synthesis inflammatory (TNF-α, IL-1β, IL-8, IL-2, IFN-γ ) and anti-inflammatory (IL-4, IL-10) cytokines by mononuclear cells of human peripheral blood by ELISA was analyzed in the article. Microorganisms Bacillus cereus (strain JP5832), isolated from the modern drug Bactisubtil, were used as control for comparison. Metabolites were obtained from a suspension of microorganisms taken in aliquots of 0.05·10⁶ or 500·10⁶ m. cl./ml, which incubated at temperatures of −5°C and 37°C. The revealed effects of the action of secondary metabolites were in a certain dependence on the temperature conditions of incubation of microorganisms, their number, and species. Metabolites of strain M3, used at a dose of 0.05·10⁶ m. cl., induced the synthesis of cytokines of nonspecific immunity (IL-1β and IL-8), the development of cell-mediated (IFN-γ) and antibody-mediated (IL-4) immune reaction, and the action of metabolites strain 8/75-1 to a greater extent was aimed at increasing the secretion of IL-2, IFN-γ by mononuclear cells. The “warm” metabolites of strain 8/75-1 and the “cold” metabolites of strain M3, taken in a dose of 500·10⁶ m. cl., have shown themselves to be active stimulants of secretion of practically the entire investigated spectrum of cytokine.

References:

1. Brushkov A. V., Mel’nikov V. P., Shchelchkova M. V., Griva G. I., Repin V. E., Brenner E. V., Tanaka M. 2011. “Biogeokhimiya merzlykh porod Tsentral’noy Yakutii” [Biogeochemistry of Permafrost of Central Yakutia]. Earth’s Cryosphere, vol. 15, no 4, pp. 90-100.
2. Budchanov Yu. I. 2008. Gormony i mediatory immunnoy sistemy. Regulyatsiya immunnogo otveta [Hormones and Mediators of the Immune System. Regulation of the Immune Response]. Tver: TSMA.
3. Kalenova L. F., Subbotin A. M., Bazhin A. S. 2013. “Vliyanie bakteriy iz mnogoletnemerzlykh porod raznogo geologicheskogo vozrasta na immunnuyu sistemu” [Influence of Bacteria from Perennial Frozen Species of Different Geological Age on the Immune System]. Journal of New Medical Technologies, no 1.
4. Kalenova L. F., Bazhin A. S., Novikova M. A. 2014. “Vliyanie metabolitov bakteriy Bacillus sp. iz vechnoy merzloty na skorost’ reparatsii kozhnoy rany” [Effects of Metabolites of Bacteria Bacillus sp. from Permafrost on Speed Repair of Skin Wound]. Journal of New Medical Technologies, vol. 21, no 4, pp. 53-61. 
5. Kalenova L. F., Subbotin A. M., Novikova M. A., Bazhin A. S. 2014. “Vliyanie temperatury inkubatsii na biologicheskuyu aktivnost’ mikroorganizmov iz vechnoy merzloty” [Effect of Incubation Temperature on the Biological Activity of Microorganisms Isolated from Permafrost]. Bulletin of Experimental Biology and Medicine, vol. 158, no 12, pp. 737-741.
6. Karaevskaya E. S., Demidov N. E., Shmelev D. G., Rivkina E. M., Bulat S. A. 2017. “Izuchenie bakterial’nykh soobshchestv mnogoletnemerzlykh porod oazisov Antarktidy metodami kul’tivirovaniya” [The Study of the Bacterial Communitis in the Antarctic Oases’ Permafrost by Means of Culturing]. Arctic and Antarctic Research, no 2 (112), pp. 27-42.
7. Kolyvanova S. S., Solovyov V. S., Kalenova L. F. 2015. “Vliyanie shtammov bakteriy roda Bacillus, vydelennykh iz mnogoletnemerzlykh porod raznogo geologicheskogo vozrasta, na immunologicheskie pokazateli laboratornykh zhivotnykh” [Effect of Bacterial Strains of the Genus Bacillus, Isolated from Permafrost of Different Geological Ages, on the Immunological Parameters of Laboratory Animals]. Russian Journal of Immunology, vol. 9 (18), no 3 (1), pp. 105-107.
8. Nikolaev Yu. A. 2011. “Autoregulaciya stressovogo otveta microorganismov” [Autoregulation of the Stress Response of Microorganisms]. Dr. Sci. (Biol.) diss. Moscow: S. N. Winogradsky Institute of Microbiology RAS.
9. Orlova T. I., Bulgakova V. G., Polin A. N. 2014. “Vtorichnyye metabolity mikroorganizmov — potentsialnyy rezerv farmatsevticheskikh preparatov” [Microbial Secondary Metabolites as Potential Reserve of Pharmaceuticals]. Antibiotics and Chemotherapy, no 3-4, рр. 39-44.
10. Shcherbakova V. A. 2018. “Anaerobnye bakterii i arkhei v mnogoletnemerzlykh otlozheniyakh Arktiki” [Anaerobic Bacteria and Archaea in Arctic Permafrost]. Dr. Sci. (Biol.) diss. Moscow: S. N. Winogradsky Institute of Microbiology RAS.
11. El-Registan G. I., Mulyukin A. L., Nikolaev Yu. A., Suzina N. E., Galchenko V. F., Duda V. I. 2006. “Adaptogennye funktsii vnekletochnykh autoregulyatorov mikroorganizmov” [Adaptogenic Functions of Extracellular Autoregulators of Microorganisms]. Microbiology, vol. 75, no 4. pp. 446-456.
12. Erin K., Shiner А., Kendra P. 2005. “Interkingdom Signaling: Deciphering the Language of Acylhomoserine Lactones”. Microbiology Reviews, vol. 29. pp. 935-947.
13. Levin B. R., Rozen D. E. 2006. “Non-Inherited Antibiotic Resistance”. Nature Reviews Microbiology, vol. 4. pp. 556-562. DOI: 10.1038/nrmicro1445