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Antibody to recombinant murine tumor necrosis factor (TNF) neutralizes guinea pig TNF
R Ruppel-Kerr1, J A Lemp, M H Karol
1Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, PA 15261.
Abstract:
Cotton dust has been found to cause acute pulmonary inflammation and fever in humans and in a guinea pig model of byssinosis. Following 3 h inhalation of cotton dust particles, guinea pig macrophages were found to release ex vivo a factor(s) toxic to WEHI fibrosarcoma cells. The cytotoxic factor(s) was also present in the bronchoalveolar fluid. We sought to investigate the mechanism of the inflammatory response to determine whether the factor was TNF. Antibodies to murine TNF were produced by immunizing sets of rabbits using two protocols. All animals produced anti-TNF antibodies with titers of 1/1000-1/25,000. Sera from one set of animals completely neutralized the cytotoxicity of murine TNF toward the WEHI cell line. The antisera neutralized up to 93% of the cytotoxicity of guinea pig samples but only 54% of human recombinant TNF. These results identify TNF in pulmonary tissues of guinea pigs following exposure to cotton dust. Moreover, the studies indicate that rabbit antibodies to murine TNF can be used to detect the guinea pig cytokine.
Insights
Cotton dust exposure triggers the release of a cytotoxic factor from guinea pig lungs, identified as tumor necrosis factor (TNF). This finding helps understand pulmonary inflammation and byssinosis mechanisms.
Area of Science:
- Pulmonary immunology
- Toxicology
- Cell biology
Background:
- Cotton dust inhalation causes acute pulmonary inflammation and fever in humans and guinea pigs.
- Guinea pig macrophages release a cytotoxic factor after cotton dust exposure, found in bronchoalveolar fluid.
Purpose of the Study:
- To investigate the inflammatory mechanism of cotton dust exposure.
- To determine if the cytotoxic factor released by guinea pig macrophages is tumor necrosis factor (TNF).
Main Methods:
- Production of antibodies to murine TNF by immunizing rabbits.
- Testing the neutralization capacity of anti-TNF antibodies against cytotoxic factors from guinea pig samples and human recombinant TNF.
Main Results:
- Rabbit antisera against murine TNF were successfully produced with high titers.
- The antisera neutralized a significant portion (93%) of the cytotoxicity in guinea pig samples, but only 54% of human recombinant TNF.
- These results indicate the presence of TNF in guinea pig pulmonary tissues after cotton dust exposure.
Conclusions:
- Tumor necrosis factor (TNF) is present in guinea pig lungs following cotton dust inhalation.
- Rabbit antibodies to murine TNF can effectively detect the guinea pig cytokine, aiding in the study of byssinosis.