The aim was to study the effects of a -agonist (clen-
buterol) on lipopolysaccharide (LPS)—induced cytokine
expression of bronchoalveolar cells. Four horses with
low-grade small airway diseases and four horses with re-
current airway obstruction were challenged on three
occasions with aerosols containing Salmonella LPS or a
0.9% saline solution. Before the first LPS challenge,
horses were not treated. Before the second and third
challenges, horses received intravenous clenbuterol
(Ventipulmin), 0.8 μg/kg twice daily, for 3.5 days. A 2-
week washout period was allowed between the chal-
lenges. Bronchoalveolar lavage fluid (BALF) was ob-
tained, and maximal intrapleural pressure ( P
pl
max)
was measured 6 hours after each challenge. After cyto-
logic examination was performed, the messenger RNA
expression in the BALF cell pellet of the following cy-
tokines was investigated by reverse-transcriptase poly-
merase chain reaction: tumor necrosis factor- (TNF- ),
interleukin (IL)-1 , IL-1 , IL-4, IL-6, IL-8 (CXCL8), IL-10,
tissue growth factor- (TGF- ), and interferon- (IFN- ).
Treatment with clenbuterol caused a decrease in maxi-
mal intrapleural pressure toward more normal values in
the recurrent airway obstruction group. IL-4 mRNA ex-
pression could not be demonstrated. Only IFN- and IL-
From the 1st Medical Clinic for Ungulates and Small Animals
a
and
Institute of Medical Chemistry,
b
Veterinary University of Vienna,
Veterinärplatz 1, Vienna, Austria.
Supported by Boehringer-Ingelheim.
Reprint requests: R. van den Hoven, DVM, PhD, 1st Medical Clinic
for Ungulates and Small Animals, Veterinary University of Vienna,
Veterinärplatz 1, Vienna 1210, Austria.
0737-0806/$ - see front matter
© 2004 Elsevier Inc. All rights reserved.
doi:10.1016/jevs.2004.12.001
Volume 24, Number 1 29
The Expression of Messenger RNA for Tumor
Necrosis Factor- ; Interleukins 1 , 6, 8, and 10;
Tissue Growth Factor- ; and Interferon- in
Bronchoalveolar Lavage Fluid Cells From Horses
Challenged With Lipopolysaccharide
and Treated With Clenbuterol
R. van den Hoven, DVM, PhD,
a
J. C. Duvigneau, PhD,
b
R. T. Hartl, Dipl Biol,
b
K. Riedelberger, Dr Med Vet,
a
M. Teinfalt, Dr Med Vet,
a
and M. Gemeiner, PhD
b
Scientific Papers
10 were statistically significantly different in the three
trials, indicating modulation of the LPS-induced inflam-
matory response by clenbuterol treatment. The expected
inhibition by clenbuterol on LPS-induced expression of
TNF- mRNA or stimulation of IL-10 mRNA could not
be clearly shown. However, IL-1, IL-10, and IFN-
mRNA tended to be downregulated by clenbuterol in all
horses; whereas IL-6 mRNA appeared to be downregu-
lated in horses with small airway diseases only. Despite
its limitations, this study showed that clenbuterol ap-
peared to modulate the communication between the
different cell types in equine airways. A different clen-
buterol dosing strategy, a different sample time, and a
heavier challenge might have caused more significant
changes. (J Equine Vet Sci 2004;24:29-36)
R
ecurrent airway obstruction (RAO), formerly re-
ferred to as chronic obstructive pulmonary dis-
ease, is a common disease of mature stabled
horses. An excellent short review of the disease
was recently given.
1
Airway inflammation plays an im-
portant role in the pathophysiology of the disorder. The
primary lesion is bronchiolitis, characterized by accumu-
lation of mucus and neutrophils in the airway lumen and
peribronchial infiltration with mononuclear cells, primar-
ily lymphocytes.
2
The action of neutrophilic granulocytes
is considered to be the major pathologic event in equine
RAO.
3
Cytokines, primarily chemokines, play a central
role in lower airway inflammation and are produced by a
multitude of cell types including neutrophils, macro-
phages, lymphocytes, eosinophils, mast cells, dendritic
cells, endothelial cells, and epithelial cells.
4
Specific cy-
tokines could serve as biomarkers for the involvement of
certain cell types in inflammatory and other pathologic
REFEREED