Persistent activation of nuclear factor-kappaB signaling pathway in severe uncontrolled asthma.
The transcription factor nuclear factor-kappaB (NF-kappaB) is inactive when bound to its inhibitory protein IkappaBalpha. On cell stimulation with inflammatory signals, IkappaBalpha is phosphorylated by IkappaB kinases and subsequently degraded. Freed NF-kappaB then induces expression of cytokines such as granulocyte-macrophage colony-stimulating factor, interleukin-8, and regulated upon activation, normal T cell expressed and secreted. These mediators are overexpressed in asthma and are downregulated by glucocorticoids through NF-kappaB activity repression. However, high levels of granulocyte-macrophage colony-stimulating factor, interleukin-8, and regulated upon activation, normal T cell expressed and presumably secreted are released by peripheral blood mononuclear cells isolated from patients with severe asthma despite continuous systemic glucocorticoid treatment. We report that these mediators are markedly decreased by pyrrolidinedithiocarbamate, an inhibitor of NF-kappaB activation. To further characterize the persistent NF-kappaB activation in severe asthma, we analyzed the expression of various components of this activation pathway in healthy subjects and in asthmatics with mild controlled, and moderate and severe uncontrolled disease. We found high amounts of phosphorylated IkappaBalpha characterizing the three asthmatic groups. Western blot analyses indicated that in peripheral blood mononuclear cells the IkappaB kinase beta and p65 levels were greater in moderate and severe asthmatics than in normal subjects. Electrophoretic mobility shift assay and immunocytochemistry showed a greater activation status of p65 in severe asthmatics. Our data suggest that exaggerated NF-kappaB activation perpetuates inflammatory mediators production in severe asthma.
Gagliardo
Rosalia
R
Istituto di Biomedicina e Immunologia Molecolare, Consiglio Nazionale delle Ricerche, Palermo, Via Ugo La Malfa 153, 90146 Palermo, Italy. gagliardo.r@iol.it
Chanez
Pascal
P
Mathieu
Marc
M
Bruno
Andreina
A
Costanzo
Giorgia
G
Gougat
Claire
C
Vachier
Isabelle
I
Bousquet
Jean
J
Bonsignore
Giovanni
G
Vignola
Antonio M
AM
D000328:Adult; D001249:Asthma / Q000503:physiopathology*; D018946:Chemokine CCL5 / Q000032:analysis; D005260:Female; D005938:Glucocorticoids / Q000008:administration & dosage; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000032:analysis; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000502:physiology*; D008297:Male; D008875:Middle Aged; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000502:physiology*; D011239:Prednisolone / Q000008:administration & dosage; D011759:Pyrrolidines / Q000494:pharmacology; D012720:Severity of Illness Index; D015398:Signal Transduction / Q000502:physiology*; D013859:Thiocarbamates / Q000494:pharmacology
D018946:Chemokine CCL5; D005938:Glucocorticoids; D016209:Interleukin-8; D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D011239:Prednisolone