title: Endotoxin contamination of ovalbumin suppresses murine immunologic responses and development of airway hyper-reactivity.
abstract: The reversible airway hyper-reactivity (AHR) of asthma is modeled by sensitizing and challenging mice with aerosolized ovalbumin. However, the C57BL/6 murine strain does not display the large increase in circulating IgG and IgE antibodies found in human atopy and asthma. We found that commercial ovalbumin was contaminated with lipopolysaccharide (LPS) in amounts sufficient to fully activate endothelial cells in an in vitro assay of the first step of inflammation. Desensitization of TLR4 by LPS pretreatment suppressed the inflammatory effect of ovalbumin. The presence of LPS was occult, because it does not require serum presentation and, like the LPS of Salmonella minnesota, was not suppressed by polymyxin B. Purified ovalbumin did not activate endothelial cells in vitro; however, endotoxin-free ovalbumin was far more effective than commercial material in stimulating IgE production and respiratory dysfunction in a C57BL/6 murine model of AHR. Moreover, endotoxin-free ovalbumin induced lung inflammation with alveolar enlargement and destruction in a histologic pattern that differed from the changes caused by commercial, endotoxin-contaminated ovalbumin. Reconstitution of purified ovalbumin with S. minnesota LPS decreased lung inflammation, decreased changes in lung function, and suppressed anti-ovalbumin antibody production. We conclude endotoxin contaminates ovalbumin preparations and that endotoxin co-administration with the ovalbumin antigen creates a state of tolerance in a murine model of AHR. Co-exposure to endotoxin and antigen occurs in humans through organic dusts, so murine models of AHR may reflect the clinical situation, but models based on commercial ovalbumin do not accurately reflect the effect of protein antigen alone on animal physiology.
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, UT 84112-5330, USA.
authors.forename: Junji
authors.initials: J
authors.lastname: Watanabe
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Miyazaki
authors.forename: Guy A
authors.initials: GA
authors.lastname: Zimmerman
authors.forename: Kurt H
authors.initials: KH
authors.lastname: Albertine
authors.forename: Thomas M
authors.initials: TM
authors.lastname: McIntyre
mesh_terms: D000402:Airway Obstruction / Q000139:chemically induced*; D000818:Animals; D001249:Asthma / Q000139:chemically induced; D004195:Disease Models, Animal; D004340:Drug Contamination; D004730:Endothelium, Vascular / Q000187:drug effects; D006801:Humans; D007108:Immune Tolerance; D007109:Immunity / Q000187:drug effects*; D007136:Immunoglobulins / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D010047:Ovalbumin / Q000008:administration & dosage / Q000494:pharmacology* / Q000592:standards; D011014:Pneumonia / Q000139:chemically induced / Q000276:immunology / Q000473:pathology; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D007136:Immunoglobulins; D008070:Lipopolysaccharides; D010047:Ovalbumin