Jill A. Poole, MD, Neil E. Alexis, PhD, Conrad Parks, BS, Amy K

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Presentation transcript:

Repetitive organic dust exposure in vitro impairs macrophage differentiation and function Jill A. Poole, MD, Neil E. Alexis, PhD, Conrad Parks, BS, Amy K. MacInnes, BS, Martha J. Gentry-Nielsen, PhD, Paul D. Fey, PhD, Lennart Larsson, PhD, Diane Allen-Gipson, PhD, Susanna G. Von Essen, MD, MPH, Debra J. Romberger, MD Journal of Allergy and Clinical Immunology Volume 122, Issue 2, Pages 375-382.e4 (August 2008) DOI: 10.1016/j.jaci.2008.05.023 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cell-surface marker expression in MDMs. After 1 week, control and ODE MDMs demonstrated mCD14 and CD16 expression by means of flow cytometry. HLA-DR, CD80, and CD86 expression was diminished with ODE versus control MDMs. A, Representative histogram of 4 separate studies. B, Mean ± SEM of the MFI (n = 4). ∗Statistically significant, P < .05). Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Cell-surface marker expression in MDMs. After 1 week, control MDMs were compared with MDMs/early exposed (A; n = 3) and MDMs conditioned with heat-inactivated dust (B; n = 3), 10 ng/mL LPS (C; n = 4), 4 μg/mL PGN (D; n = 4), combination LPS plus PGN (E; n = 4), and endotoxin-depleted ODE (F; n = 3). Values are expressed as means ± SEMs of the MFI. ∗Statistically significant, P < .05. Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Phagocytosis of IgG-opsonized Saccharomyces cerevisiae zymosan bioparticles. Phagocytosis was diminished in ODE-treated MDMs compared with that seen in control MDMs (n = 4, P = .001). A, Representative dot plot of rightward shift in fluorescence from 4 separate studies. B, Fold change in MFI (proportion of cells in the zymosan-exposed population at 60 minutes compared with cells exposed for 0 minutes). ∗Statistically significant, P < .05. Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Phagocytosis of IgG-opsonized zymosan bioparticles in MDMs. Fold change in MFI on the y-axis (proportion of cells in the zymosan-exposed population at 60 minutes compared with cells exposed for 0 minutes) with MDMs/early ODE exposed (A; n = 3) and MDMs conditioned with heat-inactivated dust (B; n = 3), LPS (C; n = 3), PGN (D; n = 3), LPS plus PGN (E; n = 3), and endotoxin-depleted ODE (F; n = 3) is shown. ∗Statistically significant, P < .05. NS, Nonsignificant, P > .05). Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Secretion of cytokines/chemokines in MDMs at 1 week after restimulation with ODE 1% for 5 hours. TNF-α (A), IL-6 (B), and IL-10 (C) secretion was dampened in ODE MDMs compared with that seen in control MDMs (n = 5, P < .05). CXCL8 (D) expression was not significantly altered (n = 5). Mean results are presented per 5 × 105 cells ± SEMs. ∗Statistically significant, P < .05). Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

A, Schematic representation of experimental protocol showing elutriated monocytes cultured with GM-CSF (1000 IU/mL) with and without low-concentration ODE (0.1%) for 6 days. B, Monocytes cultured with GM-CSF and ODE (0.1%) for 48 hours and washed and maintained in culture for the remaining 4 days (MDMs/early ODE exposed). Macrophages rested for 18 hours before experimental assays. Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Cytokine secretion in MDMs at 1 week after restimulation with ODE (1%) for 5 hours. TNF (A) and IL-6 (B) secretion was dampened in all conditioned MDMs compared with that seen in control MDMs, except for heat-inactivated ODE MDMs (n = 3). Mean results are presented as the percentage of ODE-induced cytokine secretion in control MDMs ± SEM. ∗Statistically significant, P < .05). NS, Not significant. Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Cytokine secretion in MDMs at 1 week after restimulation with LPS (100 ng/mL; A and B) and PGN (40 μg/mL; C and D). TNF (Fig E2, A and C) and IL-6 (Fig E2, B and D) secretion was dampened in all conditioned MDMs compared with that seen in control MDMs (n = 3). Mean results are presented as the percentage of LPS- or PGN-induced cytokine secretion in control MDMs ± SEM. ∗Statistically significant, P < .05). Journal of Allergy and Clinical Immunology 2008 122, 375-382.e4DOI: (10.1016/j.jaci.2008.05.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions