Effects of in vitro exposure to hay dust on the gene expression of chemokines and cell-surface receptors in primary bronchial epithelial cell cultures established from horses with chronic recurrent airway obstruction.
OBJECTIVE
To examine effects of in vitro exposure to solutions of hay dust, lipopolysaccharide (LPS), or beta-glucan on chemokine and cell-surface receptor (CSR) gene expression in primary bronchial epithelial cell cultures (BECCs) established from healthy horses and horses with recurrent airway obstruction (RAO).


METHODS
BECCs established from bronchial biopsy specimens of 6 RAO-affected horses and 6 healthy horses.


METHODS
5-day-old BECCs were treated with PBS solution, hay dust solutions, LPS, or beta-glucan for 6 or 24 hours. Gene expression of interleukin (IL)-8, chemokine (C-X-C motif) ligand 2 (CXCL2), IL-1beta, toll-like receptor 2, toll-like receptor 4, IL-1 receptor 1, and glyceraldehyde 3-phosphate dehydrogenase was measured with a kinetic PCR assay.


RESULTS
Treatment with PBS solution for 6 or 24 hours was not associated with a significant difference in chemokine or CSR expression between BECCs from either group of horses. In all BECCs, treatment with hay dust or LPS for 6 hours increased IL-8, CXCL2, and IL-1beta gene expression > 3-fold; at 24 hours, only IL-1beta expression was upregulated by > 3-fold. In all BECCs, CSR gene expression was not increased following any treatment. With the exception of a 3.7-fold upregulation of CXCL2 in BECCs from RAO-affected horses (following 6-hour hay dust treatment), no differences in chemokine or CSR gene expression were detected between the 2 groups. At 24 hours, CXCL2 gene expression in all BECCs was downregulated.


CONCLUSIONS
Epithelial CXCL2 upregulation in response to hay dust particulates may incite early airway neutrophilia in horses with RAO.
Ainsworth
Dorothy M
DM
Department of Clinical Sciences, Cornell University, Ithaca, NY 14853, USA.
Matychak
Marybeth
M
Reyner
Claudia L
CL
Erb
Hollis N
HN
Young
Jean C
JC
D000821:Animal Feed; D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D018925:Chemokines / Q000235:genetics / Q000378:metabolism*; D004391:Dust / Q000276:immunology*; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology*; D006734:Horse Diseases / Q000276:immunology*; D006736:Horses; D008070:Lipopolysaccharides; D008297:Male; D029424:Pulmonary Disease, Chronic Obstructive / Q000276:immunology / Q000662:veterinary; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*
D018925:Chemokines; D004391:Dust; D008070:Lipopolysaccharides; D011956:Receptors, Cell Surface