engleski

Tumor associated glycoprotein-72 affects the balance between pro-inflammatory and anti-inflammatory cyto/chemokine production in CD1a+ dendritic cells

Problem: Possible binding, endocytosis, and influence of Tumor Associated Glycoprotein-72 (TAG-72) on cytokine and chemokine production in CD1a+ dendritic cells were investigated. Design & methods: Normal human 6-10 weeks old pregnancy decidua was enzimatically digested by colagenase IV and centrifuged on gradient density in order to obtain decidual mononuclear cells (DMC). DMC immediately after isolation were untreated or pretreated with TAG-72 (50, 100 or 200 U/ml) or mannan (50, 100 or 200  g/ml) for 30 min on ice and then CD209 and CD206 were labeled using antibody directed toward their carbohydrate recognition domains or FITC dextran endocytosis was measured by flow cytometry. Intracellular IFN- , IL-4, IL-10, IL-15 and IL18 cytokines were detected in CD1a+ dendritic cells (DC) from DMC suspensions recovered after 18-hours culture in the medium only or in the presence of TAG-72 (200 U/ml). The chemokines CXCL10, CCL3, CCL17 and CCL22 were investigated in supernatants of LPS matured, monocyte derived dendritic cells which were cultured with or without TAG-72, as well as in TAG-72 treated or untreated enriched decidual CD1a+ supernatants by enzyme linked immunosorbent assay (ELISA). Mann-Whitney U test was used to determine statistical significance between two groups of interest. Results: TAG-72, as well as mannan (positive control) significantly reduced anti-CD206 and anti-CD209 mAb binding and FITC dextran endocytosis on a dose dependent manner in early pregnancy decidual CD1a+ DC. The addition of TAG-72 in the suspension of DMC minced substantially the percentage of IL-15 and IFN- expressing CD1a+ DC, as well as mean fluorescence intensity (MFI) for IL-15 and IL-18 in CD1a+ cells. IL-4 and IL-10 expression was not affected by TAG-72. CXCL10 secretion (approximately 40 ng/ml) was four times higher then CCL17 secretion of enriched decidual CD1a+ cells cultured in the medium only and it became balanced in the presence of TAG-72. Monocyte derived DC cultured in the medium only secreted Th1 oriented (CCL3, CXCL10) and Th2 oriented (CCL17, CCL22) chemokines after terminal LPS maturation. Secretion of CXCL10 significantly decreased in monocyte derived DC cultured in the presence of TAG-72. Conclusions: Decidual CD1a+ cells are able to bind and internalize TAG-72 by carbohydrate recognition domains of CD206 and CD209 receptors and consequently change cytokine/chemokine pattern production toward supporting Th2 oriented cells. Acknowledgement: We gratefully acknowledge the financial support provided by the Croatian Ministry of Science (Grants No. 0620402-0376 and 0620402-0377) and the EMBIC project, European FP6, NoE, LSHM-CT-2004-512040.

cyto/chemokine; dendritic cells; Tumor Associated Glycoprotein-72

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