Peripheral blood and airway tissue expression of transforming growth factor β by neutrophils in asthmatic subjects and normal control subjects Hong Wei.

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Presentation transcript:

Peripheral blood and airway tissue expression of transforming growth factor β by neutrophils in asthmatic subjects and normal control subjects Hong Wei Chu, MD, John B. Trudeau, BS, Silvana Balzar, MD, Sally E. Wenzel, MD Journal of Allergy and Clinical Immunology Volume 106, Issue 6, Pages 1115-1123 (December 2000) DOI: 10.1067/mai.2000.110556 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Compared with normal control subjects, asthmatic subjects had increased numbers of TGF-β+ cells (A) , neutrophils (B) , and TGF-β+ neutrophils (C) in the endobronchial biopsy tissues. Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Representative photomicrographs of neutrophil elastase and TGF-β1,2,3 immunostaining in 2 consecutive sections of an asthmatic endobronchial biopsy tissue. A, Neutrophil elastase staining; B, TGF-β1,2,3 staining. The cell positive for both neutrophil and TGF-β1,2,3 is indicated with an arrow (original magnification, 400×). Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 TGF-β immunostaining on peripheral blood neutrophils of an asthmatic subject. A, Diff-Quick staining showing the purity of neutrophils; B, TGF-β1,2,3 staining; C, TGF-β1 staining; D, negative control neutrophil section without TGF-β antibody incubation (original magnification: A, 1000×; B-D, 400×). Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Compared with normal control subjects, asthmatic subjects demonstrated higher levels of TGF-β1 in the supernatant of purified peripheral blood neutrophils after a 6-hour incubation in the media. Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Confocal microscopic images of a double-stained peripheral blood neutrophil from an asthmatic subject. A, Normarski (no fluorescent excitation); B, neutrophil elastase staining (FITC); C, TGF-β1 staining (rhodamine); D, overlay of B and C. Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 6 RT-PCR analysis of TGF-β1 and β-actin mRNA in peripheral blood neutrophils: ethidium bromide–stained agarose gel. A, TGF-β1. Lane 1, positive control; lane 2, asthmatic subject; lane 3, normal control subject; lane 4, 100-bp DNA ladder. B, β-Actin. Lane 1, Positive control; lane 2, asthmatic subject; lane 3, normal control subject; lane 4, 100-bp DNA ladder. Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 7 Detection of TGF-β1 and β-actin mRNA by in-cell RT-PCR in peripheral blood neutrophils of an asthmatic subject. A, TGF-β1 without nuclear Fast Red counterstaining. The blue-purple color indicates the mRNA signal. B, No signal was observed in in-cell PCR negative control where TGF-β1 PCR primers were omitted. The nuclei of the cells were counterstained with Fast Red. C, TGF-β1 with nuclear Fast Red counterstaining. D, Colocalization of TGF-β1 mRNA and neutrophil elastase immunostaining. The nuclei of the cells were not counterstained with Fast Red. The blue-purple color indicates the signal of TGF-β mRNA, and the brown-red color indicates the neutrophil elastase staining. E, β-actin with nuclear Fast Red counterstaining (original magnification, 1000×). Journal of Allergy and Clinical Immunology 2000 106, 1115-1123DOI: (10.1067/mai.2000.110556) Copyright © 2000 Mosby, Inc. Terms and Conditions