Fetal human airway smooth muscle cell production of leukocyte chemoattractants is differentially regulated by fluticasone.
BACKGROUND
Adult human airway smooth muscle (ASM) produce cytokines involved in recruitment and survival of leukocytes within airway walls. Cytokine generation by adult ASM is glucocorticoid-sensitive. Whether developing lung ASM produces cytokines in a glucocorticoid-sensitive fashion is unknown.


METHODS
Cultured fetal human ASM cells stimulated with TNF-α (0-20 ng/ml) were incubated with TNF-α receptor-blocking antibodies, fluticasone (1 and 100 nm), or vehicle. Supernatants and cells were assayed for the production of CCL5, CXCL10, and CXCL8 mRNA and protein and glucocorticoid receptor phosphorylation.


RESULTS
CCL5, CXCL10, and CXCL8 mRNA and protein production by fetal ASM cell was significantly and dose-dependently following TNF-α treatment. Cytokine mRNA and protein production were effectively blocked by TNF-α R1 and R2 receptor neutralizing antibodies but variably inhibited by fluticasone. TNF-α-induced TNF-R1 and R2 receptor mRNA expression was only partially attenuated by fluticasone. Glucocorticoid receptor phosphorylation at serine (Ser) 211 but not at Ser 226 was enhanced by fluticasone.


CONCLUSIONS
Production of CCL5, CXCL10, and CXCL8 by fetal ASM appears to involve pathways that are both qualitatively and mechanistically distinct to those described for adult ASM. The findings imply developing ASM has potential to recruit leukocyte into airways and, therefore, of relevance to childhood airway diseases.
Department of Infection, Immunity and inflammation, University of Leicester, Leicester, UK.
Helen
H
Pearson
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota.
Rodney D
RD
Britt
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota.|Department of Anesthesiology, Mayo Clinic, Rochester, Minnesota.
Christine M
CM
Pabelick
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota.|Department of Anesthesiology, Mayo Clinic, Rochester, Minnesota.
Y S
YS
Prakash
Department of Infection, Immunity and inflammation, University of Leicester, Leicester, UK.|Institute of Lung Health, Glenfield Hospital Leicester, Leicester, UK.
Yassine
Y
Amrani
Department of Infection, Immunity and inflammation, University of Leicester, Leicester, UK.|Institute of Lung Health, Glenfield Hospital Leicester, Leicester, UK.
Hitesh C
HC
Pandya
D000906:Antibodies / Q000494:pharmacology; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000235:genetics / Q000378:metabolism; D054357:Chemokine CXCL10 / Q000235:genetics / Q000378:metabolism; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D016207:Cytokines / Q000276:immunology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D000068298:Fluticasone / Q000494:pharmacology*; D005865:Gestational Age; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000187:drug effects* / Q000196:embryology / Q000276:immunology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D010766:Phosphorylation; D011965:Receptors, Glucocorticoid / Q000819:agonists / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D012694:Serine; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000906:Antibodies; C483024:CCL5 protein, human; C114753:CXCL10 protein, human; C508606:CXCL8 protein, human; D018946:Chemokine CCL5; D054357:Chemokine CXCL10; D016207:Cytokines; D005938:Glucocorticoids; D016209:Interleukin-8; D011965:Receptors, Glucocorticoid; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; C506235:TNFRSF1A protein, human; C415902:TNFRSF1B protein, human; D014409:Tumor Necrosis Factor-alpha; D012694:Serine; D000068298:Fluticasone