Measurement of pro-inflammatory and anti-inflammatory cytokines in exhaled breath condensates*1

1054 Measurement of Pro-inflammatory and Anti-inflammatoryCytokines in Exhaled Breath Condensates

P. F. Kao, C. L. Liebeler, M. N. Blumenthal; Pulmonary, Allergy and Crit-ical Care, University of Minnesota, Minneapolis, MN.RATIONALE: Asthma is a chronic, inflammatory respiratory disease.Exhaled markers of inflammation measured noninvasively may revealinformation on the changes seen with different stages of asthma as pro-inflammatory and anti-inflammatory components are hypothesized tocontribute to the pathological changes manifested in all stages of asthma.METHODS: Recruited individuals included 17 subjects with asthma and11 control subjects. Each person was given standardized pulmonary func-tion tests and exhaled breath condensates (EBC) were collected. Inflam-matory and anti-inflammatory markers including TNF, IL-1, IL-4, IL-10and IFN were collected and measured with a highly sensitive chemilumi-nescent enzyme immunoassay (ELISA) using a Bio-Tek Precision 2000Automated Pipetting System. Luminometer was used to read the plates.The cytokines were measured at picograms/ml. Regression analysis wasperformed on the background corrected RLUs (relative light units) fromthe standard curve and unknown sample concentrations are interpolated.Reproducible results were seen.RESULTS: The analysis of markers of EBC reveals no definite trends inthe pro-inflammatory cytokines including TNF, IL-1, IL-4 and the anti-inflmmatory cytokine IL-10 and IFN in asthmatic subjects. Many prob-lems were identified which may influence the results including medica-tions. The detection of picograms/ml of cytokines and studying theirratios will enhance our understanding of their additive, synergistic andantagonistic effects.CONCLUSIONS: We have demonstrated the ability to reproduciblymeasure inflammatory and anti-inflammatory cytokines in EBC. Thisshould allow us to further clarify their role in the development of asthma.Funding: NIH-NHLBI

J ALLERGY CLIN IMMUNOL Abstracts S289VOLUME 113, NUMBER 2

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