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Maturation of mast cell progenitors to mucosal mast cells during allergic pulmonary inflammation in mice.

Bankova LG, Dwyer DF, Liu AY, Austen KF, Gurish MF - Mucosal Immunol (2014)

Bottom Line: The resident tracheal CMCs had higher SSC, FcɛRI, and Kit and lower β7-integrin expression than the MMCs.By histology, the MMCs follow similar kinetics to the flow cytometry-identified mature MMCs and are notably persistent for >42 days.Steroid treatment reduced inflammation and MCp influx but had no effect on established MMCs.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT
In contrast to resident constitutive mast cells (CMCs), mucosal MCs (MMCs) appear in the lung and trachea of sensitized mice only following inhalation challenge. We monitored the influx and maturation of MCs by their expression of Kit, FcɛRI, β7-integrin and side scatter (SSC) by flow cytometry. Influx of MC progenitors (MCps) (FcɛRI(lo), Kit(int), β7(hi), and SSC(lo)) peaks 1 day after challenges and subsides to baseline by day 7 after challenge. The mature MMCs appear as a distinct population on day 7 and peak at day 14 with higher SSC and FcɛRI expression, but lower β7 and Kit expression. A distinct transitional population is present between 1 and 7 days after challenge. Maturation occurs more rapidly in the trachea. The resident tracheal CMCs had higher SSC, FcɛRI, and Kit and lower β7-integrin expression than the MMCs. By histology, the MMCs follow similar kinetics to the flow cytometry-identified mature MMCs and are notably persistent for >42 days. Steroid treatment reduced inflammation and MCp influx but had no effect on established MMCs. Thus, changes in SSC, FcɛRI, and Kit together with the expression of αE/α4:β7-integrins characterizes the development of induced MMCs from MCps and distinguishes them from resident CMCs in the trachea and large airways.

No MeSH data available.


Related in: MedlinePlus

Induction of 3 populations of MCs in lung of BALB/c mice 1 to 14 days after challenges. BALB/c mice were sensitized and either not challenged (NC) or challenged for 7 consecutive days with aerosolized antigen. The lung cells were isolated and analyzed by FACS on D1, D7, and D14 after the challenges. (a) Mean leukocyte cell counts (left panel) and mean number of FcεR1+Kit+ MCs/106 CD45+CD3-CD19-CD11b-cells in the lung, determined by FACS (right panel). (b) Representative dot plots of lung cells from NC and challenged mice analyzed for FcεR1 and Kit. Three different FcεRI+Kit+ populations are designated and colored as follows: FcεRIloKitint – green, FcεRIintKitint – red and FcεRIintKitlo – blue. (c) Concentration (number/106 CD45+ leukocytes) of lung MC lineage populations (FcεR1+Kit+) colored as in (b). Bar graphs represent means ± SEMs from 5 experiments with 7-15 mice/group. * p < 0.05, **p < 0.01, ***p < 0.001
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Figure 1: Induction of 3 populations of MCs in lung of BALB/c mice 1 to 14 days after challenges. BALB/c mice were sensitized and either not challenged (NC) or challenged for 7 consecutive days with aerosolized antigen. The lung cells were isolated and analyzed by FACS on D1, D7, and D14 after the challenges. (a) Mean leukocyte cell counts (left panel) and mean number of FcεR1+Kit+ MCs/106 CD45+CD3-CD19-CD11b-cells in the lung, determined by FACS (right panel). (b) Representative dot plots of lung cells from NC and challenged mice analyzed for FcεR1 and Kit. Three different FcεRI+Kit+ populations are designated and colored as follows: FcεRIloKitint – green, FcεRIintKitint – red and FcεRIintKitlo – blue. (c) Concentration (number/106 CD45+ leukocytes) of lung MC lineage populations (FcεR1+Kit+) colored as in (b). Bar graphs represent means ± SEMs from 5 experiments with 7-15 mice/group. * p < 0.05, **p < 0.01, ***p < 0.001

Mentions: To investigate the influx of MCps and their maturation in real time in the lungs of sensitized and aerosolized antigen (Ag)-challenged mice, we analyzed the CD45+, non–B, non–T cells for expression of Kit (CD117) and FcεRI. We used a mixture of anti-FcεRI and anti-IgE antibodies to detect occupied and unoccupied FcεRI on MCs because bound IgE can interfere with detection of FcεRIα by the MAR1 mAb.17 Compared to sensitized, non-challenged (NC) mice, the number of leukocytes harvested from the lungs (the inflammatory cell infiltrate) increases significantly to a peak at D1 post-challenge and declines to baseline levels by D14 (Figure 1a). At the same time, the increase in the concentration of MCs/106 CD45+ cells, peaks at D1 and persists on a plateau to D14 even though the inflammatory cell infiltrate, measured as number of leukocytes harvested, has returned to baseline.


Maturation of mast cell progenitors to mucosal mast cells during allergic pulmonary inflammation in mice.

Bankova LG, Dwyer DF, Liu AY, Austen KF, Gurish MF - Mucosal Immunol (2014)

Induction of 3 populations of MCs in lung of BALB/c mice 1 to 14 days after challenges. BALB/c mice were sensitized and either not challenged (NC) or challenged for 7 consecutive days with aerosolized antigen. The lung cells were isolated and analyzed by FACS on D1, D7, and D14 after the challenges. (a) Mean leukocyte cell counts (left panel) and mean number of FcεR1+Kit+ MCs/106 CD45+CD3-CD19-CD11b-cells in the lung, determined by FACS (right panel). (b) Representative dot plots of lung cells from NC and challenged mice analyzed for FcεR1 and Kit. Three different FcεRI+Kit+ populations are designated and colored as follows: FcεRIloKitint – green, FcεRIintKitint – red and FcεRIintKitlo – blue. (c) Concentration (number/106 CD45+ leukocytes) of lung MC lineage populations (FcεR1+Kit+) colored as in (b). Bar graphs represent means ± SEMs from 5 experiments with 7-15 mice/group. * p < 0.05, **p < 0.01, ***p < 0.001
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4390399&req=5

Figure 1: Induction of 3 populations of MCs in lung of BALB/c mice 1 to 14 days after challenges. BALB/c mice were sensitized and either not challenged (NC) or challenged for 7 consecutive days with aerosolized antigen. The lung cells were isolated and analyzed by FACS on D1, D7, and D14 after the challenges. (a) Mean leukocyte cell counts (left panel) and mean number of FcεR1+Kit+ MCs/106 CD45+CD3-CD19-CD11b-cells in the lung, determined by FACS (right panel). (b) Representative dot plots of lung cells from NC and challenged mice analyzed for FcεR1 and Kit. Three different FcεRI+Kit+ populations are designated and colored as follows: FcεRIloKitint – green, FcεRIintKitint – red and FcεRIintKitlo – blue. (c) Concentration (number/106 CD45+ leukocytes) of lung MC lineage populations (FcεR1+Kit+) colored as in (b). Bar graphs represent means ± SEMs from 5 experiments with 7-15 mice/group. * p < 0.05, **p < 0.01, ***p < 0.001
Mentions: To investigate the influx of MCps and their maturation in real time in the lungs of sensitized and aerosolized antigen (Ag)-challenged mice, we analyzed the CD45+, non–B, non–T cells for expression of Kit (CD117) and FcεRI. We used a mixture of anti-FcεRI and anti-IgE antibodies to detect occupied and unoccupied FcεRI on MCs because bound IgE can interfere with detection of FcεRIα by the MAR1 mAb.17 Compared to sensitized, non-challenged (NC) mice, the number of leukocytes harvested from the lungs (the inflammatory cell infiltrate) increases significantly to a peak at D1 post-challenge and declines to baseline levels by D14 (Figure 1a). At the same time, the increase in the concentration of MCs/106 CD45+ cells, peaks at D1 and persists on a plateau to D14 even though the inflammatory cell infiltrate, measured as number of leukocytes harvested, has returned to baseline.

Bottom Line: The resident tracheal CMCs had higher SSC, FcɛRI, and Kit and lower β7-integrin expression than the MMCs.By histology, the MMCs follow similar kinetics to the flow cytometry-identified mature MMCs and are notably persistent for >42 days.Steroid treatment reduced inflammation and MCp influx but had no effect on established MMCs.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT
In contrast to resident constitutive mast cells (CMCs), mucosal MCs (MMCs) appear in the lung and trachea of sensitized mice only following inhalation challenge. We monitored the influx and maturation of MCs by their expression of Kit, FcɛRI, β7-integrin and side scatter (SSC) by flow cytometry. Influx of MC progenitors (MCps) (FcɛRI(lo), Kit(int), β7(hi), and SSC(lo)) peaks 1 day after challenges and subsides to baseline by day 7 after challenge. The mature MMCs appear as a distinct population on day 7 and peak at day 14 with higher SSC and FcɛRI expression, but lower β7 and Kit expression. A distinct transitional population is present between 1 and 7 days after challenge. Maturation occurs more rapidly in the trachea. The resident tracheal CMCs had higher SSC, FcɛRI, and Kit and lower β7-integrin expression than the MMCs. By histology, the MMCs follow similar kinetics to the flow cytometry-identified mature MMCs and are notably persistent for >42 days. Steroid treatment reduced inflammation and MCp influx but had no effect on established MMCs. Thus, changes in SSC, FcɛRI, and Kit together with the expression of αE/α4:β7-integrins characterizes the development of induced MMCs from MCps and distinguishes them from resident CMCs in the trachea and large airways.

No MeSH data available.


Related in: MedlinePlus