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Interleukin-5 messenger RNA expression in peripheral blood CD4+ cells in asthma  Christopher K.W. Lai, DM, FRCP(E), Alice S.S. Ho, MRCP, Christopher H.S.

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Presentation on theme: "Interleukin-5 messenger RNA expression in peripheral blood CD4+ cells in asthma  Christopher K.W. Lai, DM, FRCP(E), Alice S.S. Ho, MRCP, Christopher H.S."— Presentation transcript:

1 Interleukin-5 messenger RNA expression in peripheral blood CD4+ cells in asthma 
Christopher K.W. Lai, DM, FRCP(E), Alice S.S. Ho, MRCP, Christopher H.S. Chan, MRCP, Janet Tang, BSc, Joseph C.K. Leung, MSc, Kar N. Lai, DSc, FRCP  Journal of Allergy and Clinical Immunology  Volume 97, Issue 6, Pages (June 1996) DOI: /S (96) Copyright © 1996 Mosby, Inc. Terms and Conditions

2 FIG. 1 Demonstration of PCR products of β-actin and IL-5 mRNA from unstimulated CD4+ T cells of three subjects with stable asthma (SA) and six with acute severe asthma (AA) by agarose gel electrophoresis. β-Actin and IL-5 bands are seen in the positive control lane (+) (cDNA for β-actin and IL-5 from Clontech Laboratory) but not in the negative control (-) (reagent control in which RNA was replaced by DEPC-dH2O). Note absence of readily detectable bands for IL-5 in the mRNA from unstimulated cells. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

3 FIG. 2 Demonstration of PCR products of β-actin and IL-5 mRNA from stimulated CD4+ T cells from five patients with acute asthma (AA) before (D1) and after systemic steroid therapy (D7), five patients with stable asthma (SA), and five normal subjects (N) by agarose gel electrophoresis. M, φX174/Hae III; +, positive control (cDNA for β-actin and IL-5 from Clontech Laboratory); -, negative control (reagent control in which RNA was replaced by DEPC-dH2O). Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

4 FIG. 3 IL-5 to β-actin mRNA ratio in patients with acute and stable asthma and in normal healthy volunteers. All 21 patients with acute asthma from whom samples were obtained on days 1 and 7 demonstrated a reduction in IL-5 expression a week after systemic corticosteroid therapy. Bar indicates median values. *p = 0.034; not significant after Bonferonni correction. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

5 FIG. 4 Kinetics of amplification of β-actin and IL-5 cDNAs. The data are the mean values of 10 specimens—five from patients with acute asthma before the commencement of systemic steroid therapy and five from patients with stable asthma. After 30 PCR cycles, as described in the Methods section, the amplification was still in the linear part of the curves for both cDNAs, suggesting that amplification was not saturable under our experimental conditions. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

6 FIG. 5 Serum ECP concentrations in patients with acute and stable asthma and normal healthy volunteers. All but one of the 21 patients with acute asthma from whom samples were obtained twice demonstrated a reduction in ECP level. Bar indicates median values. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

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