Repeat organic dust exposure–induced monocyte inflammation is associated with protein kinase C activity Jill A. Poole, MD, Todd A. Wyatt, PhD, Susanna.

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Repeat organic dust exposure–induced monocyte inflammation is associated with protein kinase C activity Jill A. Poole, MD, Todd A. Wyatt, PhD, Susanna G. Von Essen, MD, MPH, John Hervert, BS, Conrad Parks, BS, Tracy Mathisen, BS, Debra J. Romberger, MD Journal of Allergy and Clinical Immunology Volume 120, Issue 2, Pages 366-373 (August 2007) DOI: 10.1016/j.jaci.2007.04.033 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Dose-dependent secretion of TNF-α by THP-1 cells to a 5-hour challenge with complete DE, endotoxin-depleted DE, and boiled DE. Mean results are presented as concentration (pg/mL) per 1 × 105 cells with SE bars shown (n = 3). ∗Statistically significant (P < .05). NS, Nonsignificant. Journal of Allergy and Clinical Immunology 2007 120, 366-373DOI: (10.1016/j.jaci.2007.04.033) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Diminished secretion of TNF-α by organic dust–pretreated THP-1 cells after restimulation. A, Complete DE. B, Endotoxin-depleted DE. THP-1 cells were pretreated with or without DE for 18 hours, then restimulated for 5 hours. Mean results are presented as concentration (pg/mL) per 105 cells with SE bars shown (n = 3). ∗Statistically significant (P < .05). PreRx, Pretreatment. Journal of Allergy and Clinical Immunology 2007 120, 366-373DOI: (10.1016/j.jaci.2007.04.033) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Secretion of inflammatory mediators in human PBMs with peptidoglycan (PG), LPS, and DE. Monocytes pretreated with PG, LPS, and DE for 18 hours, and then restimulated for 5 hours. A, TNF-α. B, IL-6. C, CXCL8. D, IL-10. Mean results are presented as concentration (pg/mL) per 105 cells with SE bars shown (n = 8 separate donors). ∗Statistically significant (P < .05). PreRx, Pretreatment. Journal of Allergy and Clinical Immunology 2007 120, 366-373DOI: (10.1016/j.jaci.2007.04.033) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Organic dust–induced PKC isoform activity. THP-1 cells were incubated with or without complete dust extract (5%) for 15, 30, 60, 180, and 360 minutes, and then assayed for PKCα, PKCδ, PKCɛ, and PKCζ. Mean results shown as kinase activity fold increase compared with control media with SE bars (n = 3). ∗Statistically significant compared with control; P < .05. Journal of Allergy and Clinical Immunology 2007 120, 366-373DOI: (10.1016/j.jaci.2007.04.033) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 PKC isoform activation in THP-1 cells pretreated with complete DE after restimulation. Cells pretreated with or without DE for 18 hours, then rechallenged for 30 minutes. Mean results shown as kinase activity fold increase compared with control media with SE bars (n = 3). Statistical values are based on comparison with cells without pretreatment. ∗Statistically significant (P < .05). PreRx, Pretreatment. Journal of Allergy and Clinical Immunology 2007 120, 366-373DOI: (10.1016/j.jaci.2007.04.033) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Effects of PKC isoform inhibitors on DE (5%)–induced secretion of TNF-α in THP-1 cells. PKC inhibitors: α inhibitor, Gö 6976; ɛ inhibitor, Ro 31-8220; δ inhibitor, Rottlerin; ζ inhibitor, myristoylated PKCζ. Mean results presented as the percentage of dust-alone TNF-α secretion with SE bars shown (n = 3). ∗Statistically significant (P < .05). INH, Inhibitor. Journal of Allergy and Clinical Immunology 2007 120, 366-373DOI: (10.1016/j.jaci.2007.04.033) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions