Activation of macrophages by gliadin fragments: isolation and characterization of active peptide.
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.
Tucková
Ludmila
L
Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic. tuckova@biomed.cas.cz
Novotná
Jana
J
Novák
Petr
P
Flegelová
Zuzana
Z
Kveton
Tomás
T
Jelínková
Lenka
L
Zídek
Zdenek
Z
Man
Petr
P
Tlaskalová-Hogenová
Helena
H
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
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