Gene Expression Pattern of Peyer's Patch Lymphocytes Exposed to Kagocel Suggests Pattern-Recognition Receptors Mediate Its Action.

Alexander A Andreev-Andrievskiy,Olga Yu Frolova,Boris A Rudoy,Victoria S Scherbakova,Vladimir G Nesterenko,Roman A Zinovkin,Yuriy G Kazaishvili,Mikhail A Mashkin

doi:10.3389/fphar.2021.679511

Abstract

Kagocel is a synthetic carboxymethylcellulose derivative copolymerized with gossypol. Clinical data evidence its safety and efficiency for the treatment of flu and other viral infections via enhancement of interferon production. The gut-associated lymphoid tissue seems a likely site of kagocel action. The study was aimed to investigate the molecular mechanisms of its action using murine Peyer’s patches lymphocytes as a test system and the cytokines production and gene expression patterns as the primary outcomes. The Peyer’s patches lymphocytes isolated from BALB/c mice were stimulated with concanavalin A, or, to mimic viral infection, with a combination of concanavalin A and TLR3 ligand poly I:C. After 24 h of stimulation the cells were treated with saline, 30, 100, or 300 μg/ml of kagocel, or, as positive controls, 300 μg/ml oats b-D-glucan or 300 μg/ml lentinan. After 24 and 72 h of incubation with these drugs cytokines production was analyzed with ELISA and gene expression pattern was investigated using nCounter Inflammation panel chips followed by bioinformatics analysis. Expression of genes involved in the inflammatory response, antiviral defense, lymphocytes survival and proliferation (C1qa, C2, C3, Ccl21a, Il11, Il1b, Il23a, Il5, Ltb4r2, Alox15, Pla2g4a, Ptger1, Mapkapk5, Hras, Ifna1, Tlr2, Mrc1, Mx2) was upregulated in kagocel-treated Peyer’s patches lymphocytes. A list of plausible transcription factors (CEBPs, IRF, NFκB, RXR, Stat, Tead4, and ZSCAN) and master-regulators has been identified (cIAP, CIKS, dock9, MEKK1, FXR, IKK, IRAK, TRAF, dsRNA:TLR3:TRIF). The changes in gene expression pattern and the outcomes of bioinformatics analysis suggest that pattern recognition receptors, TLRs and dectin-1, are the key mediators of kagocel immunomodulatory action, with the possible involvement of interferon autocrine loop. The genes upregulated with kagocel include diverse components of the innate immune defense system.

Highlights

The breakdown products of bacteria, fungi, and viruses trigger a rapid reaction of immune cells and some other cell types in the body as a part of an innate immune response (Kieser and Kagan, 2017)
Kagocel is a synthetic copolymer of modified carboxymethylcellulose and a natural polyphenol, gossypol, which was designed at the Gamaleya Research Institute of Epidemiology and Microbiology and further marketed as an oral interferon inductor in Russia and CIS countries by Nearmedic, LLC
Analysis of variance revealed significant effects of time and mitogen stimulation applied on the concentration of all the cytokines studied, except for IL10, which was not affected by the type of mitogen (Figure 1)

Summary

Introduction

The breakdown products of bacteria, fungi, and viruses trigger a rapid reaction of immune cells and some other cell types in the body as a part of an innate immune response (Kieser and Kagan, 2017) Among these breakdown products, (pathogen-associated molecular patterns, PAMPs) polysaccharides of the bacterial and fungal cell wall upon binding to toll-like receptors (Iwasaki and Medzhitov, 2004), dectin-1 receptors, and possibly other receptor types (Brown, 2005) induce inflammation, interferon production, promote immune cells survival and proliferation (Iwasaki and Medzhitov, 2004; Brown, 2005; Kieser and Kagan, 2017). No adverse effects were identified in the chronic and reproduction toxicity studies with moderate doses of kagocel (Borovskaya, 2017)

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