Interleukin 10 Induces the Expression of Membrane-Bound HLA-G and the Production of Soluble HLA-G on HeLa CCL-2 Cells

Abstract

BACKGROUND: Interleukin-10 is a cytokine that has a pleiotropic effect on the immune system and inflammation. IL-10 can contribute to the anti-tumour immune response by increasing HLA-G expression. AIM: This study aimed to determine the effect of IL-10 induction on membrane HLA-G expression and soluble HLA-G production of HeLa CCL-2 cells. METHODS: HeLa CCL-2 cells were cultured in the well plate and divided into 4 groups consist of 1 control group that was not induced by IL-10 and 3 treatment groups that were induced by IL-10 500 ng/ml, 1000 ng/ml and 2000 ng/ml respectively. All groups were incubated for 48 hours in a 37°C incubator at 5% CO2 atmospheric pressure. HLA-G measurements were carried out both in cell lysate and cell culture supernatant using ELISA and in membrane-bound using immunofluorescence method. The expression of HLA-G in membrane-bound calculated using the ImageJ application. Data obtained were analysed by ANOVA and LSD test. RESULTS: In the control group, the HLA-G level in the culture supernatant was higher than in cell lysate (p = 0.000), as well as in all treatment groups (p = 0.000). There were significant differences between the treatment group (p = 0.000) and within the treatment group (p = 0.000) at HLA levels. The highest expression of HLA-G in HeLa cell membranes found in cell culture induced by IL-10 concentrations of 500 ng/ml, i.e., 59.28 AU in view. HLA-G membrane expression in the IL-10 1000 ng/ml induced group was significantly different from all treatment groups (p = 0.000). CONCLUSION: HeLa CCL-2 cells express HLA-G on the membrane and release dissolved HLA-G without induction of IL-10 although IL-10 induction augments the presence and the production of HLA-G in HeLa CCl-2 cells. Introduction Interleukin-10, also called human cytokine synthesis inhibitory factor, is a cytokine secreted by monocytes which has a pleiotropic effect on the immune system and inflammation. Most leukocytes secrete IL-10 to a certain level. The highest secretion of IL-10 comes from monocytes and their mature forms: macrophages and dendritic cells. Specific granulocytes and agranulocytes: eosinophil and NK cells, and small lymphocytes: T and B cells, also release IL-10 but at lower levels [1]. Human leukocyte antigen (HLA)-G is HLA class 1