Analysis of the Biological Properties of Blood Plasma Protein with GcMAF Functional Activity

Abstract

The main problem related to the studies focusing on group-specific component protein-derived macrophage-activating factor (GcMAF) is the lack of clarity about changes occurring in different types of macrophages and related changes in their properties under the effect of GcMAF in various clinical conditions. We analyzed the antitumor therapeutic properties of GcMAF in a Lewis carcinoma model in two clinical conditions: untreated tumor lesion and tumor resorption after exposure to Karanahan therapy. GcMAF is formed during site-specific deglycosylation of vitamin D3 binding protein (DBP). DBP was obtained from the blood of healthy donors using affinity chromatography on a column with covalently bound actin. GcMAF-related factor (GcMAF-RF) was converted in a mixture with induced lymphocytes through the cellular enzymatic pathway. The obtained GcMAF-RF activates murine peritoneal macrophages (p < 0.05), induces functional properties of dendritic cells (p < 0.05) and promotes in vitro polarization of human M0 macrophages to M1 macrophages (p < 0.01). Treatment of whole blood cells with GcMAF-RF results in active production of both pro- and anti-inflammatory cytokines. It is shown that macrophage activation by GcMAF-RF is inhibited by tumor-secreted factors. In order to identify the specific antitumor effect of GcMAF-RF-activated macrophages, an approach to primary reduction of humoral suppressor activity of the tumor using the Karanahan therapy followed by macrophage activation in the tumor-associated stroma (TAS) was proposed. A prominent additive effect of GcMAF-RF, which enhances the primary immune response activation by the Karanahan therapy, was shown in the model of murine Lewis carcinoma. Inhibition of the suppressive effect of TAS is the main condition required for the manifestation of the antitumor effect of GcMAF-RF. When properly applied in combination with any chemotherapy, significantly reducing the humoral immune response at the advanced tumor site, GcMAF-RF is a promising antitumor therapeutic agent that additively destroys the pro-tumor properties of macrophages of the tumor stroma.

The Macrophage Activator GcMAF-RF Enhances the Antitumor Effect of Karanahan Technology through Induction of M2–M1 Macrophage Reprogramming

“…To analyze the events occurring in the tumor of experimental animals treated with Karanahan technology solely and in combination with GcMAF-RF [ 11 ], the results of four experimental series reported in [ 1 , 2 ] were used: May 2020 (No. 1), November 2020 (No.…”

“…The protocol for obtaining group-specific macrophage activator (GcMAF-RF) is described in detail in [ 1 , 11 ]. GcMAF-RF is the subject of industrial property of “ACTIVATOR MAF” LLC.…”

“…Tumor-bearing mice received similar injections of saline. The procedure used to prepare GcMAF-RF was thoroughly described in [ 1 ].…”

“…Studies [ 1 , 2 ] have characterized a new antitumor potential of Karanahan technology—the ability to induce an antitumor immune response. Karanahan technology is described in detail in [ 2 ]; it is time-ordered treatment of tumor-bearing mice with a cross-linking cytostatic cyclophosphamide and a complex double-stranded DNA agent.…”

“…Based on the functional properties of GcMAF-RF, its antitumor effect may be associated with the activation of immune responses by inhibiting the immunosuppressive capabilities of myeloid cells, such as tumor-associated macrophages and myeloid-derived suppressor cells (MDSCs), in the tumor microenvironment; the most important element of the synergistic contribution of GcMAF-RF to tumor lysis is related to its capability of inhibiting the synthesis of protumor factors suppressing tumor-reactive iimmune responses in the tumor and its stroma. At this point, M2 macrophages become the target of GcMAF-RF, and repolarization of the M2 protumor phenotype to the M1 antitumor or M0 not-activated phenotype occurs [ 1 ].…”