Flow Cytometric Analysis of Macrophages and Cytokines Profile in the Bronchoalveolar Lavage Fluid in Patients with Lung Cancer

SIMPLE SUMMARY: Macrophages are an integral part of the tumor microenvironment, playing a role in immunoregulation. We investigated the antigenic and cytokine macrophages profile derived from bronchoalveolar lavage fluid in lung affected by cancer (cBALF) and healthy lung (hBALF) of 36 patients. Macrophages markers: CD206, CD163, CD80, CD86, CD40, Arginase-1, and CD68 were evaluated by flow cytometry. Cytokines (IL-1 RA, IL-6, IL-10, TNF-α, IL-1β, IL-12, IL-23, and TGF-β) profile was analyzed. There was higher median proportion of macrophages in cBALF than in hBALF. The population of macrophages presented immunophenotype: cCD68+(bright) CD206+(bright) CD163+(bright) CD80+ CD86+ CD40+(bright) CD45+ cArginase+. We observed some trends in the expression of the analyzed antigens in clBALF and hlBLAF. High concentrations of IL-1RA and IL-6 and correlations between pro- and anti-inflammatory cytokines in cBALF and hBALF supernatants were found. We expanded knowledge of the macrophages polarization, their diversity and unique properties based on the antigenic pattern and cytokine profile. ABSTRACT: Macrophages play an important role in the suppression and activation of immune anti-cancer response, but little is known about dominant macrophage phenotype in the lung cancer environment, evaluated by bronchoalveolar lavage fluid (BALF). The aim of this study was to characterize macrophages in BALF from a lung affected by cancer (cBALF) and a healthy lung (hBALF) of the same patient regarding their individual macrophage polarization and selected cytokines profile. A total of 36 patients with confirmed lung cancer were investigated. Macrophages markers: CD206 CD163 CD80 CD86 CD40 CD45, Arginase-1, and CD68 were evaluated by flow cytometry. Cytokines (IL-1 RA, IL-6, IL-10, TNF-α, IL-1β, IL-12, IL-23, and TGF-β) profile was analyzed. There was higher median proportion of macrophages in Cbalf than in Hbalf. The population of macrophages presented immunophenotype: Ccd68+(bright) CD206+(bright) CD163+(bright) CD80+ CD86+ CD40+(bright) CD45+ cArginase+. We observed some trends in the expression of the analyzed antigens in clBALF and hlBLAF. The highest concentrations of IL-1RA and IL-6 were in Cbalf and Hbalf supernatant. There were the correlations between pro- and anti-inflammatory cytokines. The findings showed that macrophages include a diverse and plastic group with the presence of different antigens and cytokines, and determining the target phenotype is a complex and variable process.