Background: We previously observed that A-gliadin peptide P31-43 induces effects similar to Epidermal growth factor (EGF) both in cultured cell lines and in enterocytes from celiac disease (CD) patients. The effect is mediated by delayed EGF degradation and prolonged EGF receptor (EGFR) activation in endocytic vesicles due to P31-43 mediated interference with endocytic maturation.Objective: To test P31-43 effects on IL15 induction at level of transcription, translation, intracellular trafficking and its role in P31-43 induced proliferation.Methods: Semi-quantitative and real time PCR investigated P31-43 effects on IL15 mRNA levels. Protein levels and distribution was analyzed by Facs, Elisa and immunofluorescence. Stat5 and IL15R alfa activation has been examined by WB. BrdU (Bromodeoxiuridine) analyzed proliferation.Results: In Caco 2 cells P31-43 does not increases IL15 mRNA levels. IL15 protein was found increased only on the cells surface together with markers of recycling vesicles, as trasferrin and Lamp2, implying a P31-43-mediated interference with IL15 vesicular trafficking. IL15 is linked to the receptor, is not dependent on new protein synthesis and functions as a growth factor. Stat 5 and the IL15 receptor alfa (IL15Ra) are activated after P31-43 treatment. Anti- IL15 blocking antibodies can prevent P31-43 induced increase of proliferation in Caco2 cells and in enterocytes of biopsies from CD patients.Conclusion: P31-43 increases IL15 levels on the surface of Caco2 cells interfering with its vesicular trafficking. IL15 so presented functions as a growth factor and P31-43 induced proliferation can be prevented by inhibitors of both EGFR and IL15 pathway activation.

Gliadin peptide P31-43 presents in trans IL15.

AURICCHIO, SALVATORE
2008

Abstract

Background: We previously observed that A-gliadin peptide P31-43 induces effects similar to Epidermal growth factor (EGF) both in cultured cell lines and in enterocytes from celiac disease (CD) patients. The effect is mediated by delayed EGF degradation and prolonged EGF receptor (EGFR) activation in endocytic vesicles due to P31-43 mediated interference with endocytic maturation.Objective: To test P31-43 effects on IL15 induction at level of transcription, translation, intracellular trafficking and its role in P31-43 induced proliferation.Methods: Semi-quantitative and real time PCR investigated P31-43 effects on IL15 mRNA levels. Protein levels and distribution was analyzed by Facs, Elisa and immunofluorescence. Stat5 and IL15R alfa activation has been examined by WB. BrdU (Bromodeoxiuridine) analyzed proliferation.Results: In Caco 2 cells P31-43 does not increases IL15 mRNA levels. IL15 protein was found increased only on the cells surface together with markers of recycling vesicles, as trasferrin and Lamp2, implying a P31-43-mediated interference with IL15 vesicular trafficking. IL15 is linked to the receptor, is not dependent on new protein synthesis and functions as a growth factor. Stat 5 and the IL15 receptor alfa (IL15Ra) are activated after P31-43 treatment. Anti- IL15 blocking antibodies can prevent P31-43 induced increase of proliferation in Caco2 cells and in enterocytes of biopsies from CD patients.Conclusion: P31-43 increases IL15 levels on the surface of Caco2 cells interfering with its vesicular trafficking. IL15 so presented functions as a growth factor and P31-43 induced proliferation can be prevented by inhibitors of both EGFR and IL15 pathway activation.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11588/362284
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