Effect of macrophage-activating factor (GcMAF-RF) upon ex vivo polarization of macrophages, activation of dendritic cells and production of cytokines by human whole blood cells

2021 ◽  
Vol 23 (2) ◽  
pp. 257-274
Author(s):  
S. S. Kirikovich ◽  
E. V. Levites ◽  
E. V. Dolgova ◽  
A. S. Proskurina ◽  
G. S. Ritter ◽  
...  

This article is the second communication in a series of articles devoted to the effects of a domestic preparation of macrophage-activating factor (GcMAF-RF) and assessment of its biological properties. The aim of this work was to study the effect of the GcMAF-RF upon M0 → M1 polarization of macrophages (Mph), and activation of the professional properties of ex vivo generated antigen-presenting dendritic cells (DC), as well as on ex vivo production of pro-inflammatory (TNFα, IL-1β, IL-6, IFNγ, IL-17, IL-18) and anti-inflammatory (TGF-β, IL-4, IL-10) cytokines, growth factors (IL-2, GM-CSF, G-CSF, VEGF) and chemokines (MCP, IL-8) by the whole blood cells from healthy donors. Mph and DC were generated from the monocytes (3 to 5×106 /ml) derived from adherent fraction of peripheral blood mononuclear cells (MNC) of healthy donors. Granulocyte/macrophage colony-stimulating factor (rhGM-CSF) was used to obtain Mph, whereas DC production was induced by GM-CSF and interferon-α. To provide M1 polarizing signals, bacterial lipopolysaccharide (LPS from E. coli 0114:B4) was used in controls. In experimental series, GcMAF-RF was added 48 h before the end of culture. The stimulating effect of the obtained Mph and DC upon cell proliferation was assessed in allogeneic mixed culture of leukocytes (alloMLC) using radiometric technique, by 3 H-thymidine incorporation. The influence index (IR) of Mph or DC upon allo-SCL was calculated as the ratio of the proliferative response of MNCs in the presence of Mph, or DC to the level of spontaneous MNC proliferation. To determine the cytokine production by human whole blood cells ex vivo, peripheral blood samples from 3 donors with two replicate GcMAF-RF preparations were used, at a total of 6 points. All variants of the study were carried out with mitogen-activated and non-activated blood cells. The cytokine content was determined by the ELISA assays. The effects of GcMAF-RF were quantified as a fold increase (FI), i.e., the ratio of cytokine production in the presence of GcMAF-RF to the level of their spontaneous production. It was shown that the GcMAF-RF preparation was as effective, as lipopolysaccharide (LPS), the standard Mph and DC activator which induces polarization of differentiated M0-macrophages into M1 cells and final maturation of DCs, manifesting by a significant increase in their allo-stimulatory activity in a mixed leukocyte culture (allo-MLC). Moreover, GcMAF-RF stimulates production of numerous cytokines and chemokines (TNFα, IL-1β, IL-6, IL-18, IL-4, IL-10, GM-CSF, G-CSF, VEGF, IL-8), by blood cells (granulocytes, lymphocytes, monocytes), thus indicating direct participation of the macrophage activator GcMAF-RF in various immune processes. The domestic GcMAF-RF drug induces polarization of macrophages M0 → M1, final maturation of DCs and allostimulating activity of Mf and DCs, and is also able to effectively stimulate circulating blood cells to synthesize cytokines/chemokines with pro-inflammatory and immunoregulatory activities. 

Maturitas ◽  
1997 ◽  
Vol 26 (1) ◽  
pp. 63-71 ◽  
Author(s):  
S.X. Zheng ◽  
Y. Vrindts ◽  
M. Lopez ◽  
D. De Groote ◽  
P.F. Zangerle ◽  
...  

2017 ◽  
Vol 3 (1) ◽  
pp. 1344115 ◽  
Author(s):  
Esvet Akbas ◽  
Fatih Caglar Celikezen ◽  
Hasan Turkez ◽  
Ozlem Ozdemir ◽  
Adem Ruzgar ◽  
...  

10.12737/5025 ◽  
2014 ◽  
Vol 8 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Никифоров ◽  
V. Nikiforov ◽  
Терехов ◽  
I. Terekhov ◽  
Хадарцев ◽  
...  

The study discusses the effect on the functional state of the whole blood cells of low-intensity microwave radiation. On the model of intercellular interactions in the conditions of spontaneous cell activity and by stimulating cells complex mitogen, the influence of microwave radiation 1000 MHz on production by whole blood cells interleukins: IL-1β, 2, 6, 8, 10, 12, 13, 17A, TNF-alpha, IFN-g, RAIL-1, and soluble forms of toll-like receptor 1, 2, 4, 6 types was studied. The research results show that a single 45 minutes of exposure of microwave radiation of 0.05 PPM microwatt/cm2 has the ability to enhance mitogen-stimulated production of RAIL-1 by 38,7% (p=0.03), to reduce the induced production of IL-1β by 26,3% (p=0,037), IL-8 by 56,2% (p=0.022) and to increase oppressed segment complex mitogen production of IL-10 by 27,8% (p=0,041). The microwave radiation increased spontaneous and mitogen-stimulated expression of TLR, especially TLR1 during the initial low level.


2015 ◽  
Vol 68 (4) ◽  
pp. 821-827 ◽  
Author(s):  
Fatih Çağlar Çelikezen ◽  
Başak Toğar ◽  
Fatma Betül Özgeriş ◽  
Mehmet Sait İzgi ◽  
Hasan Türkez

2020 ◽  
Vol 54 (3) ◽  
pp. 91-95
Author(s):  
Dijana Topalović ◽  
Lada Živković ◽  
Ninoslav Đelić ◽  
Vladan Bajić ◽  
Biljana Spremo-Potparević

Objective. Inosine 5'-monophosphate dehydrogenase (IMPDH) activity in cancer cells is increased. Tiazofurin selectively inhibits the activity of IMPDH, and it has been granted for the treatment of different cancers and new viral diseases. Its widespread use was limited because exposure to tiazofurin under certain circumstances was found to have a higher frequency of severe non-hematologic toxicity. Therefore, the objective of this study was to examine genotoxic action and inducement of DNA damage of tiazofurin using the comet assay. Methods. The ability of tiazofurin to induce DNA damage was evaluated using single-cell gel electrophoresis (SCGE) technique/comet assay. Human whole blood cells were exposed to three final concentrations of tiazofurin (1µM/mL, 2 µM/mL, and 5 µM/mL) for 30 min in vitro. Results. Our results indicate that tiazofurin produced a significant level of DNA damage on whole blood cells after 30 min of exposure vs. control. All tested concentrations were significantly comet-forming, in a concentration-dependent manner. Conclusion. Our investigation on the tiazofurin-treated cells and their relationship to the formation of DNA damage demonstrated that the genotoxic effect was induced after exposure to tiazofurin under described conditions.


2021 ◽  
Author(s):  
Nilgun Yakubogullari ◽  
Ali Cagir ◽  
Erdal Bedir ◽  
Duygu Sag

Astragaloside VII (AST VII), a plant triterpenoid saponin isolated from Astragalus species, shows promise as vaccine adjuvant, as it supports a balanced Th1/Th2 immune response. However, the underlying mechanisms of its adjuvant activity have not been defined. Here we investigated the impact of AST VII and its newly synthesized semi-synthetic analogs on human whole blood cells, as well as on mouse bone marrow-derived dendritic cells (BMDCs). Cells were stimulated with AST VII and its derivatives in the presence or absence of LPS or PMA/ionomycin and the secretion of cytokines and the expression of activation markers were analyzed by ELISA and flow cytometry, respectively. AST VII and its analogs increased the production of IL-1b; in PMA/ionomycin stimulated human whole blood cells. In LPS-treated mouse BMDCs, AST VII increased the production of IL-1b; and IL-12, and the expression of MHC II, CD86, and CD80. The strength of the IL-1b; boost correlated directly with the hydrophobicity of the AST VII compounds. In mixed leukocyte reaction, AST VII and derivatives increased the expression of the activation marker CD44 on mouse CD4+ and CD8+ T cells. In conclusion, AST VII and its derivatives strengthen pro-inflammatory responses, support dendritic cell maturation, and T cell activation in vitro. Our results provide insights into the mechanisms of the adjuvant activities of AST VII and its analogs, which will be instrumental to improve their utility as vaccine adjuvant.


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