In vitro effects of paleobacteria exometabolites from permafrost soils on development of immune response

We have tested the effects of metabolites from paleobacteria Bacillus cereus, strain 875 TS, from Pleistocene-Holocene permafrost rocks aiming to assess the mechanisms and features of in vitro immune response in the culture of human peripheral blood mononuclear cells. It was found that paleobacterial exometabolites significantly activate monocyte differentiation into subpopulations of intermediate (CD14⁺CD16⁺) and non-classical (CD14^loCD16⁺) monocytes, effector CD4⁺ and CD8⁺T lymphocytes, with changes of early (CD69), intermediate (CD25) and late (HLA-DR) activation markers, Treg differentiation (CD3⁺CD4⁺CD25^hiCD127^-). These metabolites also stimulated the synthesis of IFNγ and IL-4 cytokines as compared with control levels. The differential influence of paleobacterial exometabolites upon immunomodulatory activity depended on temperature regimen of their production, i.e., “cold” (obtained from bacteria during their cultivation at 5 °C), “medium-temperature” (22 °C) and “heat” (37 °C) regimens. “Cold” metabolites stimulate predominantly the immune response mechanisms with proinflammatory activity, i.e., differentiation of intermediate CD14⁺CD16⁺ monocytes, increased differentiation activity of CD8⁺T lymphocytes, and synthesis of IFNγ. “Warm” metabolites stimulate mostly the immune response mechanisms with anti-inflammatory activity, namely, differentiation of non-classical CD14^loCD16⁺ monocytes, increased differentiation activity of CD4⁺T lymphocytes and secretion of IL-4. Another distinctive feature is the ratio of pro- and anti-inflammatory mechanisms, which does not depend on the type of exometabolites. Thus, during the first three days of cell cultivation, the differentiation activity of CD8⁺T lymphocytes prevails over the differentiation of CD4⁺T lymphocytes, and the level of IFNγ secretion exceeds the IL-4 amounts. On the third day, there is a significant increase in the Treg level, which is accompanied by a tendency to normalize the balance between IFNγ (Th1) and IL-4 (Th2) by the seventh day. We have observed a clear effect of Treg (CD3⁺CD4⁺CD25^hiCD127^-) on the strength and duration of the immune response. The increase in Treg levels occurs moderately and transiently, which, on the one hand, prevents the excessive development of proinflammatory mechanisms, and on the other hand, does not lead to the development of long-term immune suppression. An increase in the Treg level on days 1-3 is accompanied by a decreased activity of monocyte differentiation into the subsets and the synthesis of proinflammatory IFNγ cytokine. Considering that one of the main functions of induced Treg is the suppression of systemic inflammatory, autoimmune and allergic diseases, increase in their activity under the influence of exometabolites of paleobacteria Bacillus cereus strain 875 TS may serve as a basis for the development of new biopreparations for treatment of a wide range of diseases.

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