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Blood fibrocytes are recruited during acute exacerbations of chronic obstructive pulmonary disease through a CXCR4-dependent pathway  Isabelle Dupin,

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Presentation on theme: "Blood fibrocytes are recruited during acute exacerbations of chronic obstructive pulmonary disease through a CXCR4-dependent pathway  Isabelle Dupin,"— Presentation transcript:

1 Blood fibrocytes are recruited during acute exacerbations of chronic obstructive pulmonary disease through a CXCR4-dependent pathway  Isabelle Dupin, PhD, Benoit Allard, PhD, Annaig Ozier, MD, PhD, Elise Maurat, MSc, Olga Ousova, PhD, Eva Delbrel, MSc, Thomas Trian, PhD, Hoang-Nam Bui, MD, Claire Dromer, MD, Olivier Guisset, MD, Elodie Blanchard, MD, Gilles Hilbert, MD, PhD, Frédéric Vargas, MD, PhD, Matthieu Thumerel, MD, Roger Marthan, MD, PhD, Pierre-Olivier Girodet, MD, PhD, Patrick Berger, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 137, Issue 4, Pages e7 (April 2016) DOI: /j.jaci Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Study design. Numbers of patients who were included and had their fibrocyte counts quantified. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Increased blood fibrocyte counts during COPD exacerbation. A and B, Circulating fibrocytes (CD45+Col1+ cells) expressed as a percentage of PBMCs (Fig 2, A) and counts in blood (Fig 2, B) from control subjects (Cont, n = 40), patients with nonexacerbating COPD (NEx, n = 9), and patients with exacerbating COPD (V1, n = 54) are shown. *P < .05, **P < .01, and ***P < .001, nonparametric Kruskal-Wallis test. C and D, CD45+CD34+Col1+ cells expressed as a percentages of PBMCs (Fig 2, C) and counts in blood (Fig 2, D) from control subjects (Cont, n = 29), patients with nonexacerbating COPD (NEx, n = 8), and patients with exacerbating COPD (V1, n = 41). *P < .05, nonparametric Kruskal-Wallis test. Fig 2, A-D, Medians are represented as horizontal lines. E and F, Comparison of fibrocytes (CD45+Col1+ cells) in patients with exacerbating COPD at the time of exacerbation (V1) and 2 months after exacerbation resolution (V2). **P < .01, Wilcoxon matched pairs test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Consequences of increased fibrocyte percentages on survival and lung function in patients with COPD. A, Kaplan-Meier survival analysis comparing patients with exacerbating COPD and greater than 28% CD45+Col1+ cells among PBMCs measured at the time of exacerbation (n = 6; black curve) with patients with less than 28% CD45+Col1+ cells among PBMCs (n = 36; gray curve). B-D, Relationships between FEV1 (Fig 3, B), FVC (Fig 3, C), Pao2 (Fig 3, D), and the percentage of CD45+Col1+ cells in PBMCs in patients with exacerbating COPD at V2. Correlation coefficient (r) and significance level (P value) were obtained by using nonparametric Spearman analysis. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Analysis of fibrocyte marker expression by means of flow cytometry. Expression of CXCR4 (A and B), CCR2 (C and D), CCR3 (E and F), and CCR7 (G and H) in fibrocytes from control subjects (Cont) and patients with exacerbating COPD (V1). Fibrocytes are CD45+Col1+ cells. Results were expressed as percentages of PBMCs (Fig 4, A, C, E, and G) and counts in blood (Fig 4, B, D, F, and H). *P < .05 and **P < .01, Mann-Whitney tests. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Analysis of fibrocyte chemotaxis. A, Migration of fibrocytes from control subjects (n = 8, gray bars) and patients with exacerbating COPD (n = 6, black bars) in response to plasma from patients with exacerbating COPD in the presence or absence of 25 μg/mL plerixafor. *P < .05, paired t test. B, Plasma CXCL12-α levels in individual subjects. Cont, Control subjects; NE, patients with nonexacerbating COPD; V1, patients with exacerbating COPD during an AECOPD; V2, patients with exacerbating COPD in the stable state. C, Migration of fibrocytes from control subjects (n = 8, gray lines) and patients with exacerbating COPD (n = 5, black lines) in response to CXCL12. **P < .01, 2-way ANOVA with Bonferroni tests. D, Migration of fibrocytes from control subjects (n = 5) and patients with exacerbating COPD (n = 7) in response to CXCL12 in the presence or absence of 25 μg/mL plerixafor. *P < .05, paired t test. Results were expressed as means ± SEMs (Fig 5, A, C, and D) or with symbols indicating individual subject values and horizontal gray lines representing medians (Fig 5, B). Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E1 Identification of fibrocytes by using flow cytometry and phase-contrast microscopy. Representative dot plots of flow cytometry for fibrocyte quantification from a patient with COPD exacerbation. A, Total NANT cell population selected on unstained cells. B, Isotype control for CD45 set on unstained cells. C, Positive population for CD45. D, Isotype control for collagen-1 set on a CD45-gated population. E, Positive population for both CD45 and collagen-1. FITC, Fluorescein isothiocyanate; FSC-A, forward scatter; SSC-A, side scatter. F, Representative image of cultured blood fibrocytes taken with a phase-contrast microscope, showing their morphology as spindle-shaped adherent cells. Bar = 20 μm. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E2 Percentage of CD45+Col1+ cells among PBMCs of patients with exacerbating COPD at V2 with 1 or no unscheduled visits (n = 15) or with 2 or more unscheduled visits (n = 16) the year before V1. Medians are represented as gray horizontal lines. *P < .05, Mann-Whitney test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E3 A, Migration of fibrocytes from control subjects (n = 3, gray bars) and patients with exacerbating COPD (n = 6, black bars) in response to plasma of patients with exacerbating COPD in the presence or absence of 3.8 μg/mL SB (paired t test). B, Plasma CCL11 levels in individual subjects. Cont, Control subjects; NE, patients with nonexacerbating COPD; V1, patients with exacerbating COPD during an AECOPD; V2, patients with exacerbating COPD in the stable state. C, Migration of fibrocytes from control subjects (n = 4, gray lines) and patients with exacerbating COPD (n = 6, black lines) in response to CCL11. D, Migration of fibrocytes from control subjects (n = 4) and patients with exacerbating COPD (n = 5) in response to CCL11 in the presence or absence of 3.8 μg/mL SB P < .001, paired t test. Results are expressed as means ± SEMs (Fig E3, A, C, and D) or with symbols indicating individual subject values and horizontal gray lines representing medians (Fig E3, B). ***P < .001. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E4 Individual CXCL12-β plasma concentrations. Cont, Control subjects; NEx, patients with nonexacerbating COPD; V1, patients with exacerbating COPD during an AECOPD; V2, patients with exacerbating COPD in the stable state. Symbols indicate individual subjects, and horizontal gray lines represent medians. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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