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Published on: March 17, 2020
Effects of gliadin stimulation on bone marrow-derived dendritic cells from HLA-DQ8 transgenic MICE
R Ciccocioppo1, M Rossi, I Pesce
1Center for the Study and Cure of Coeliac Disease, IRCCS San Matteo Hospital Foundation, University of Pavia, Italy. rachele.ciccocioppo@unipv.it
Background And Aim:
Gliadin presentation by HLA-DQ2/8 molecules to T cells plays a crucial role in triggering the inflammatory cascade in coeliac disease. We aimed to study the immunological effects of gliadin stimulation on dendritic cells (DCs) from HLA-DQ8 transgenic and BALB/c mice.
Methods:
Bone marrow-derived DCs were stimulated with alpha-chymotrypsin-digested gliadin or ovoalbumin (100 microg/ml). Modification of DC maturation, through HLA-DQ8 and MHC class II expression, and activation, by CD80 and CD86, was assessed by flow cytometry. The ability of pulsed and unpulsed DCs to prime T cells was evaluated by mixed leucocyte reaction. The expression of interleukin-4, -10, -12p70 and interferon-alpha, as well as of Toll-like receptor-4, -7, -8, -9 was determined by ELISA and real-time RT-PCR, respectively.
Results:
Gliadin stimulation induced DC maturation (p<0.001 in BALB/c, p<0.01 in DQ8) but not activation, whereas ovoalbumin upregulated all markers (p<0.01 for maturation and p<0.001 for activation in both DC populations). No increase of T proliferation was elicited by pulsed DCs with respect to unpulsed DCs. Only in DQ8 DCs, gliadin induced Toll-like receptor-4 (p<0.001), -7 (p<0.001), -8 (p<0.005) expression and interferon-alpha (p<0.001) secretion.
Conclusion:
Gliadin resulted unable to activate DC, but stimulated Toll-like receptor expression and interferon-alpha secretion.
