title: Activation of macrophages by gliadin fragments: isolation and characterization of active peptide.
abstract: Celiac disease, induced by dietary gluten, is characterized by mucosal atrophy and local inflammation associated with cell infiltration and activation. Unlike other food proteins, gluten and its proteolytic fragments, besides inducing a specific immune response, were shown to activate components of innate immunity and cause, e.g., direct stimulation of TNF-alpha and IL-10 and a significant rise in NO production by peritoneal macrophages. The identity of the active fragments was established by separating the peptic digest of gliadin by RP-HPLC chromatography. The purest fraction with the highest activity was analyzed by mass spectrometry, and the gliadin peptide sequence was identified as VSFQQPQQQYPSSQ. This peptide (T) and its N- and C-terminally shortened forms (A, B, C and D, E, F) were synthesized. Peptide B (FQQPQQQYPSSQ) elicited the highest TNF-alpha, IL-10, and RANTES secretion and increase in IFN-gamma-primed NO production by mouse macrophages. In contrast, C-terminally shortened peptides had a lower ability to stimulate macrophages than the native form.
authors.affiliation: Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic. tuckova@biomed.cas.cz
authors.forename: Ludmila
authors.initials: L
authors.lastname: Tucková
authors.forename: Jana
authors.initials: J
authors.lastname: Novotná
authors.forename: Petr
authors.initials: P
authors.lastname: Novák
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Flegelová
authors.forename: Tomás
authors.initials: T
authors.lastname: Kveton
authors.forename: Lenka
authors.initials: L
authors.lastname: Jelínková
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Zídek
authors.forename: Petr
authors.initials: P
authors.lastname: Man
authors.forename: Helena
authors.initials: H
authors.lastname: Tlaskalová-Hogenová
mesh_terms: D000818:Animals; D018946:Chemokine CCL5 / Q000096:biosynthesis; D005260:Female; D005903:Gliadin / Q000494:pharmacology*; D016753:Interleukin-10 / Q000096:biosynthesis; D008262:Macrophage Activation / Q000187:drug effects*; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D010446:Peptide Fragments / Q000302:isolation & purification / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D018946:Chemokine CCL5; D010446:Peptide Fragments; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D009569:Nitric Oxide; D005903:Gliadin; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse