Tag Archives: macrophages

Changes in proHB-EGF expression after functional activation of the immune system cells

T. O. Chudina1,2, A. J. Labintsev1, S. I. Romaniuk1, D. V. Kolybo1, S. V. Komisarenko1

1Palladin Institute of Biochemistry, National Academy  of Sciences of Ukraine, Kyiv;
2ESC Institute of Biology and Medicine, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: kolibo@biochem.kiev.ua

The level of proHB-EGF expression on J774, Raji, KG-1 cells derived from different types of human and mouse immune system cells under the standard in vitro culture conditions and during functional activation of these cells was investigated. Changes in the proHB-EGF expression on the cell surface were found to depend on the density of cell population, the content of fetal bovine serum in the culture medium, the effect of mitogenic factors – bacterial lipopolysaccharide, an inactive full-size form of diphtheria toxin (CRM197) and recombinant soluble HB-EGF – rsHB-EGF. The results obtained are important for the understanding of the functional role of proHB-EGF receptor on the surface of macrophage-like cells and B lymphocytes and indicate the involvement of this receptor in immune response regulation in an organism.

The effect of thymic mesenchymal stromal cells on arginase activity and nitric oxide produced by mouse macrophages

R. S. Dovgiy1,2, I. S. Nikolsky3, L. M. Skivka1

1Taras Shevchenko National University of Kyiv, Ukraine;
2Institute of Gerontology, NAMS of Ukraine, Kyiv;
3State Institute of Genetic and Regenerative
Medicine NAMS of Ukraine, Kyiv;
e-mail: romandovgiy@gmail.com

Mesenchymal stromal cells (MSC) gained much attention due to their therapeutic properties, media­ted largely by anti-inflammatory action. We aimed to investigate the capacity of MSC obtained from young mice to modulate arginine metabolism of macrophages from old animals. Bone marrow cells obtained from young and aged mice were cocultivated with MSC in the presence of M-CSF. Nitric oxide production was analyzed in supernatants by Griess reaction, and arginase activity was measured in cell lysates. We have found that arginase activity was significantly lower in macrophages isolated from old mice as compared to young animals (P ˂ 0.05). Syngeneic MSC addition markedly stimulated arginase activity in macrophages from both young and aged mice (P ˂ 0.001), with greater effect in old animals. There were no significant differences in nitric oxide level between groups. In summary, there was more pronounced anti-inflammatory shift in macrophage metabolism in aged animals upon cocultivation with MSC.

Mycobacterium tuberculosis antigens MPT63 and MPT83 increase phagocytic activity of murine peritoneal macrophages

A. A. Siromolot1,2, O. S. Oliinyk2, D. V. Kolibo2,1, S. V. Komisarenko2

1Educational and Scientific Centre Institute of Biology,
Taras Shevchenko National University of Kyiv, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: saa0205@ukr.net

Macrophages (MΦ) are the most described and characterized target and host of mycobacteria. Like other cells of innate immunity MΦ have a wide range of receptor molecules which interact with different pathogen associated molecular patterns (PAMPs). Immunodominant proteins MPT63 and MPT83 that are synthesized in abundance by Mycobacterium bovis or Mycobacterium tuberculosis strains could be involved in development of tuberculosis infection. The aim of this study was to search for effects of these mycobacterial antigens on target cells. For this aim full-sized sequences of MPT83 (rMPT83full) and MPT63 antigens were cloned into plasmid pET24a(+). The increase of phagocytic activity of murine peritoneal macrophages was demonstrated, but not of macrophage-like cells from J774 cell line, which were treated by rMPT63 and rMPT83full proteins for 24 h. This effect of such antigens can be considered as a way to facilitate the consumption of mycobacterial cells by macrophages to avoid other effector mechanisms of innate and adaptive immunity.