Glucosaminylmuramyl dipeptide acid (GMDP-A) modulates intracellular signaling pathways in natural killer cells

• Svetlana V. Guryanova
• Gaponov A.M.
• Pisarev V.M.
• Yakushenko E.V.
• Tutelyan A.V.
• Alexandrov I.A.
• Tsipandina E.V.
• Ivan G. Kozlov

Abstract

Introduction. Medicines developed as derivatives of muramyl peptides have been the subject of experimental and clinical studies for several decades. The intracellular NOD2- receptor responsible for the binding of muramyl dipeptide and its derivatives remains an attractive molecular target for the development of new drugs with a known mechanism of action. GMDP-A belongs to one of the NOD2-receptor ligands. In this study signaling mechanisms initiated by GMDP-A were characterized for the first time in natural killer (NK) cells.

Material and methods. NK cells isolated by magnetic separation from peripheral blood of healthy donors were incubated with 10 pg/ml GMDP-A for 6 hours and 16 hours. The levels of genes expression were determined using the Illumina HumanHT-12v4 Expression BeadChip kit and Illumina iScan instrument (Illumina, USA). The data were statistically processed using an application software package (v 2011.1, Illumina).

Results. It was found that GMDP-A enhanced the transcription of a number of genes associated with transduction of signals from receptors, thereby ensuring the maintenance of the functional activity of NK cells. GMDP-A induced the synthesis of transcripts of the MAPK signaling pathway, VAV2 signal mediators, the CRKL signal adapter, enhanced the synthesis of transcripts of the regulatory subunit 3 (γ) phosphoinositide-3-kinase and phospholipase C B1 genes that control phospholipid metabolism, activated the cells with defects in the cellular immunity link. GMDP-A significantly increases the expression level of the following key genes associated with the control of the mechanisms of direct and antibody-mediated cytotoxic activity of NK cells: CRKL, VAV2, ZAP70, RAP1A, PLCB1, FCGR3A, the MAPK family, IFNA1, TNFRSF9, TNFSF14.

Conclusion. Experimental data justify the high potential of the GMDP-A to counteract altered recognition of the early stages of oncogenic and viral transformation by key cells of the innate immunity, the NK cells.

Keywords:natural killer cells; glucosaminyl muramyl dipeptide acid; transcriptome; MAPK signaling pathway; interferon; anti-infection immunity; anti-tumor immunity

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