Gene Expression of Subunits of the IL-12 Family Cytokines in moDCs Derived In Vitro from the Cord Blood of Children of Healthy and Allergic Mothers
Abstract
The incidence of allergic diseases is steadily increasing an urgent need to clarify the immunologic processes which occur early in life and signal an increased risk of possible future allergy development. The ratio and maturation state of DCs together with the cytokine environment are important in directing and modulating immune responses.
The maturation state (presence of CD83) of cord blood monocyte-derived dendritic cells (moDCs) 01 52 children of healthy mothers and 58 children of allergic mothers was estimated by flow cytometry. The capacity of moDCs to express genes for subunits of IL-12 family cytokines was monitored using real-time PCR and protein secretion in cell culture supernatants by ELISA.
The percentage of CD83(+) moDCs was significantly higher in the allergic group after LPS stimulation (43.11 +/- 4.41) in comparison to the healthy group (24.85 +/- 3.37). Significantly higher gene expression of subunits of IL-12 family members was observed in moDCs of children of allergic mothers, in comparison with children of healthy mothers.
The differences were evident mainly after LPS stimulation of moDCs (healthy group: p19: 3.05 +/- 1.24; p28: 14.8 +/- 6.8;p35: 1.8 +/- 0.6; p40: 8.0 +/- 3.5; EBI3: 3.0 +/- 1.2; allergic group: p19: 6.1 +/- 2.7; p28: 61.4 +/- 22.2; p35: 14.9 +/- 6.5; p40: 36.4 +/- 18.8; EBI3: 11.3 +/- 3.2), with the exception of p28, whose expression was significantly higher in the allergic group even without stimulation (healthy group: 0.28 +/- 0.12, allergic group: 0.87 +/- 0.62). No significant difference between the healthy and allergic groups was found at the protein level.
The observation of both increased presence of cell surface activation marker on moDCs and higher IL-12 family gene expression in LPS-stimulated moDCs of children of allergic mothers indicates a higher reactivity of these cells.