Related expression of arachidonate 12- and 15-lipoxygenases in animal and human lung tissue.
We examined the immunohistochemical distribution of the arachidonate 12- and 15-lipoxygenases in animal and human lung tissue using a polyclonal anti-12/15-lipoxygenase antibody. Immunoblotting of whole cell extracts from bovine and human tracheal epithelial cells or from bovine leukocytes with the antibody (raised originally against purified porcine leukocyte 12-lipoxygenase) showed immunoperoxidase staining of a single protein band (Mr = 72,000), which comigrated with purified bovine 12-lipoxygenase. The antibody also immunoprecipitated both 12- and 15-lipoxygenase activities from cytosolic fractions of bovine and human tracheal epithelial cells. Immunohistochemistry of formaldehyde-fixed and paraffin-embedded bovine (and ovine and canine) trachea using the same polyclonal antibody and an indirect biotin-avidin-peroxidase detection system demonstrated specific staining of tracheal epithelium, polymorphonuclear and mononuclear leukocytes, and perineural cells. Less intense staining of submucosal glands and blood vessels was also observed. Lung sections demonstrated that the level of lipoxygenase antigen decreased markedly by the level of the bronchi and was absent in more distal airways. A similar pattern of immunostaining was found in human lung, except that airway smooth muscle was also weakly reactive, and polymorphonuclear (neutrophilic) leukocytes were unstained (in accordance with the low 12/15-lipoxygenase activity in this cell type). We conclude that animal and human epithelial 12/15-lipoxygenases share enzymatic, antigenic, and regional distribution characteristics and may therefore possess a common function in the pulmonary airway.
Shannon
V R
VR
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Crouch
E C
EC
Takahashi
Y
Y
Ueda
N
N
Yamamoto
S
S
Holtzman
M J
MJ
D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D004848:Epithelium / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007422:Intestines / Q000201:enzymology; D008168:Lung / Q000201:enzymology*; D011233:Precipitin Tests; D012867:Skin / Q000201:enzymology; D014132:Trachea / Q000201:enzymology
D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase