The primary role in biologic activity of the neutrophil chemokines IL-8 and GRO-alpha in cultured nasal epithelial cells.
Primary nasal epithelial cells were investigated for their ability to synthesize and deliver neutrophil chemotactic factors (chemokines) following tumor necrosis factor-alpha (TNF-alpha) induction. The chemokines interleukin8 (IL-8), growth-related oncogene-alpha (GRO-alpha), epithelial cell-derived neutrophil attractant-78 (ENA-78), and granulocyte chemotactic protein-2 (GCP-2) have been detected and characterized and shown to have different potencies in the chemotaxis of neutrophils. Cultures of primary nasal epithelial cells were treated with TNF-alpha in concentrations of 20 and 200 ng/ml for 2, 8, 24, and 72 h. The chemokine protein concentrations in the supernatants of the incubations were determined by the ELISA technique. Chemokine mRNA expression in epithelial cells was also measured using the reverse transcriptase-polymerase chain reaction (RT-PCR). The biologic activity of the chemokines was identified using a three-step high-performance liquid chromatography (HPLC) technique, a bioassay involving measurement of neutrophil chemotaxis in a single Boyden chamber. Both the IL-8 and GRO-alpha proteins and their respective mRNA appear to be induced by TNF-alpha in epithelial cells. The chemotactic responsiveness of both GRO-alpha and IL-8 appears to predominate after 24 h incubation with TNF-alpha. The chemokines GCP-2 and ENA-78 were only weakly induced by TNF-alpha. The neutrophil chemokines IL-8 and GRO-alpha were synthesized in nasal epithelial cell culture induced by TNF-alpha in biologically active concentrations of 0.8 ng/ml and 1.42 ng/ml, respectively. It appears that both the IL-8 and GRO-alpha chemokines may contribute to neutrophil tissue migration in sinusitis, whereas GCP-2 and ENA-78 are of secondary importance to the chemotaxis of neutrophils in this condition.
Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital, Münster, Germany. rudack@uni-muenster.de
Claudia
C
Rudack
Steffen
S
Maune
Johannes
J
Eble
Jens-Michael
JM
Schroeder
D002478:Cells, Cultured; D054360:Chemokine CXCL1; D054365:Chemokine CXCL5; D054427:Chemokine CXCL6; D018925:Chemokines / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D019743:Chemokines, CXC / Q000378:metabolism; D002630:Chemotactic Factors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002634:Chemotaxis, Leukocyte; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D016209:Interleukin-8 / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D009297:Nasal Mucosa / Q000166:cytology; D018375:Neutrophil Activation; D009504:Neutrophils / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C475829:CXCL1 protein, human; C486263:CXCL5 protein, human; C515503:CXCL6 protein, human; D054360:Chemokine CXCL1; D054365:Chemokine CXCL5; D054427:Chemokine CXCL6; D018925:Chemokines; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha