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Measurement of pro-inflammatory and anti-inflammatory cytokines in exhaled breath condensates
Abstracts S289
1054
Measurement of Pro-inflammatory and Anti-inflammatory Cytokines in Exhaled Breath Condensates
P. F. Kao, C. L. Liebeler, M. N. Blumenthal; Pulmonary, Allergy and Critical Care, University of Minnesota, Minneapolis, MN. RATIONALE: Asthma is a chronic, inflammatory respiratory disease. Exhaled markers of inflammation measured noninvasively may reveal information on the changes seen with different stages of asthma as proinflammatory and anti-inflammatory components are hypothesized to contribute to the pathological changes manifested in all stages of asthma. METHODS: Recruited individuals included 17 subjects with asthma and 11 control subjects. Each person was given standardized pulmonary function tests and exhaled breath condensates (EBC) were collected. Inflammatory and anti-inflammatory markers including TNF, IL-1, IL-4, IL-10 and IFN were collected and measured with a highly sensitive chemiluminescent enzyme immunoassay (ELISA) using a Bio-Tek Precision 2000 Automated Pipetting System. Luminometer was used to read the plates. The cytokines were measured at picograms/ml. Regression analysis was performed on the background corrected RLUs (relative light units) from the standard curve and unknown sample concentrations are interpolated. Reproducible results were seen. RESULTS: The analysis of markers of EBC reveals no definite trends in the pro-inflammatory cytokines including TNF, IL-1, IL-4 and the antiinflmmatory cytokine IL-10 and IFN in asthmatic subjects. Many problems were identified which may influence the results including medications. The detection of picograms/ml of cytokines and studying their ratios will enhance our understanding of their additive, synergistic and antagonistic effects. CONCLUSIONS: We have demonstrated the ability to reproducibly measure inflammatory and anti-inflammatory cytokines in EBC. This should allow us to further clarify their role in the development of asthma. Funding: NIH-NHLBI
TUESDAY
J ALLERGY CLIN IMMUNOL VOLUME 113, NUMBER 2
1054
Measurement of Pro-inflammatory and Anti-inflammatory Cytokines in Exhaled Breath Condensates
P. F. Kao, C. L. Liebeler, M. N. Blumenthal; Pulmonary, Allergy and Critical Care, University of Minnesota, Minneapolis, MN. RATIONALE: Asthma is a chronic, inflammatory respiratory disease. Exhaled markers of inflammation measured noninvasively may reveal information on the changes seen with different stages of asthma as proinflammatory and anti-inflammatory components are hypothesized to contribute to the pathological changes manifested in all stages of asthma. METHODS: Recruited individuals included 17 subjects with asthma and 11 control subjects. Each person was given standardized pulmonary function tests and exhaled breath condensates (EBC) were collected. Inflammatory and anti-inflammatory markers including TNF, IL-1, IL-4, IL-10 and IFN were collected and measured with a highly sensitive chemiluminescent enzyme immunoassay (ELISA) using a Bio-Tek Precision 2000 Automated Pipetting System. Luminometer was used to read the plates. The cytokines were measured at picograms/ml. Regression analysis was performed on the background corrected RLUs (relative light units) from the standard curve and unknown sample concentrations are interpolated. Reproducible results were seen. RESULTS: The analysis of markers of EBC reveals no definite trends in the pro-inflammatory cytokines including TNF, IL-1, IL-4 and the antiinflmmatory cytokine IL-10 and IFN in asthmatic subjects. Many problems were identified which may influence the results including medications. The detection of picograms/ml of cytokines and studying their ratios will enhance our understanding of their additive, synergistic and antagonistic effects. CONCLUSIONS: We have demonstrated the ability to reproducibly measure inflammatory and anti-inflammatory cytokines in EBC. This should allow us to further clarify their role in the development of asthma. Funding: NIH-NHLBI
TUESDAY
J ALLERGY CLIN IMMUNOL VOLUME 113, NUMBER 2