Joanna J. Lukawska, MD, Lefteris Livieratos, PhD, Barbara M

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Presentation transcript:

Real-time differential tracking of human neutrophil and eosinophil migration in vivo Joanna J. Lukawska, MD, Lefteris Livieratos, PhD, Barbara M. Sawyer, Tak Lee, MD, ScD, Michael O'Doherty, MD, MD, Philip J. Blower, PhD, Martin Kofi, James R. Ballinger, PhD, Christopher J. Corrigan, MD, PhD, Gopinath Gnanasegaran, MD, Ehsan Sharif-Paghaleh, PhD, Gregory E.D. Mullen, PhD Journal of Allergy and Clinical Immunology Volume 133, Issue 1, Pages 233-239.e1 (January 2014) DOI: 10.1016/j.jaci.2013.06.031 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Example images (anterior view) of 1-minute frames from the acquired dynamic image sequence shown at 1, 5, 15, 25, and 120 minutes after injection of radiolabeled neutrophils (top row) and eosinophils (bottom row). The lung, liver, and spleen are indicated by arrows. Journal of Allergy and Clinical Immunology 2014 133, 233-239.e1DOI: (10.1016/j.jaci.2013.06.031) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Percentage mean lung TACs of labeled neutrophils and eosinophils in 4 volunteers with a fitted curve. Error bars represent ± 1 SD, with a fitted curve. Rapid clearance of eosinophils from the left and right lungs was exhibited (mean half-life, 4.16 ± 0.11 minutes). Neutrophils had significantly slower lung clearance (mean half-life, 13.72 ± 4.14 minutes). Journal of Allergy and Clinical Immunology 2014 133, 233-239.e1DOI: (10.1016/j.jaci.2013.06.031) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Mean TACs for neutrophils and eosinophils of the liver (A) and spleen (B). Error bars represent ± 1 SD. In the liver eosinophils showed initial faster uptake, a reflection of faster lung migration. There were no significant differences noted after the first 15 minutes. In the spleen eosinophils demonstrated significantly higher uptake than neutrophils (P = .014). Journal of Allergy and Clinical Immunology 2014 133, 233-239.e1DOI: (10.1016/j.jaci.2013.06.031) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Example of the distribution of radioactivity over 250 minutes for the lungs, liver, spleen, and mediastinum in a single representative healthy volunteer for purified autologous neutrophils (A) and purified autologous eosinophils (B). cps, Counts per second. Journal of Allergy and Clinical Immunology 2014 133, 233-239.e1DOI: (10.1016/j.jaci.2013.06.031) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 CD16/CD11bhigh neutrophil population established by means of flow cytometry. A, Unfixed healthy volunteer whole blood was stained with CD16 and CD11b and analyzed by means of flow cytometry. Quadrants are based on negatively stained cells and isotype controls. Cells were gated on granulocytes and analyzed for the CD16/CD11blow population (left square gate). B, The CD16/CD11bhigh neutrophil population was determined by means of incubation of unfixed healthy volunteer whole blood with GM-CSF and then analyzed by means of flow cytometry. Quadrants are based on negatively stained cells and isotype controls. Cells were gated on granulocytes and analyzed for the CD16/CD11bhigh population (right square gate). Journal of Allergy and Clinical Immunology 2014 133, 233-239.e1DOI: (10.1016/j.jaci.2013.06.031) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions