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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.


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Expression of β7 and β1 integrins and forward and side scatter characteristics of tracheal constitutive and inducible MC populations at D7 after challenges. (a) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β7 integrin expression on the 3 populations of tracheal MCs, CMCs (orange shaded), eMMCs (red) and MMCs (blue); black dotted line -isotype control. (b) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β1 (CD29) integrin expression on the 3 populations of tracheal MCs colored as in a. (c) Representative histograms of the side scatter characteristics of the 3 MC populations in trachea (colored as in a and compared with splenic MCp (grey shaded) and intraperitoneal MCs (back dotted line). (d) Forward scatter characteristics of tracheal, splenic and intraperitoneal MC populations as in c. Histograms in c and d show the mean value from 2-3 mice/group from one of 5 experiments. Bar graphs represent mean (± SEM) from 4 experiments with 5-8 mice per group *p < 0.05, **p < 0.01, ***p < 0.001.
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Figure 5: Expression of β7 and β1 integrins and forward and side scatter characteristics of tracheal constitutive and inducible MC populations at D7 after challenges. (a) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β7 integrin expression on the 3 populations of tracheal MCs, CMCs (orange shaded), eMMCs (red) and MMCs (blue); black dotted line -isotype control. (b) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β1 (CD29) integrin expression on the 3 populations of tracheal MCs colored as in a. (c) Representative histograms of the side scatter characteristics of the 3 MC populations in trachea (colored as in a and compared with splenic MCp (grey shaded) and intraperitoneal MCs (back dotted line). (d) Forward scatter characteristics of tracheal, splenic and intraperitoneal MC populations as in c. Histograms in c and d show the mean value from 2-3 mice/group from one of 5 experiments. Bar graphs represent mean (± SEM) from 4 experiments with 5-8 mice per group *p < 0.05, **p < 0.01, ***p < 0.001.

Mentions: The eMMCs transiently present on D1 in the trachea have the highest β7 integrin expression of MCs in the trachea (Figure 5a, red line and bar). The mature MMCs have less β7 integrin expression (Figure 5a, blue line and bar) than eMMCs while their β1 expression is similar (Figure 5b). The granularity by SSC is greater in MMCs than the eMMCs, which is greater than splenic MCps (Figure 5c, blue line, red line, and grey shaded area, respectively). Nonetheless, the SSC of MMCs is still lower than that of the tracheal FcεRIhiKithi CMCs, which is modest compared to peritoneal MCs (Figure 5c, orange line and black dotted line, respectively). The FSC of CMCs is lower than that of the inducible trachea eMMC and MMC populations, with the eMMCs being comparable to peritoneal MCs (Figure 5d).


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)

Expression of β7 and β1 integrins and forward and side scatter characteristics of tracheal constitutive and inducible MC populations at D7 after challenges. (a) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β7 integrin expression on the 3 populations of tracheal MCs, CMCs (orange shaded), eMMCs (red) and MMCs (blue); black dotted line -isotype control. (b) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β1 (CD29) integrin expression on the 3 populations of tracheal MCs colored as in a. (c) Representative histograms of the side scatter characteristics of the 3 MC populations in trachea (colored as in a and compared with splenic MCp (grey shaded) and intraperitoneal MCs (back dotted line). (d) Forward scatter characteristics of tracheal, splenic and intraperitoneal MC populations as in c. Histograms in c and d show the mean value from 2-3 mice/group from one of 5 experiments. Bar graphs represent mean (± SEM) from 4 experiments with 5-8 mice per group *p < 0.05, **p < 0.01, ***p < 0.001.
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Figure 5: Expression of β7 and β1 integrins and forward and side scatter characteristics of tracheal constitutive and inducible MC populations at D7 after challenges. (a) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β7 integrin expression on the 3 populations of tracheal MCs, CMCs (orange shaded), eMMCs (red) and MMCs (blue); black dotted line -isotype control. (b) Representative histograms (top panel) and mean (± SEM) net MFI (bottom panel) of the β1 (CD29) integrin expression on the 3 populations of tracheal MCs colored as in a. (c) Representative histograms of the side scatter characteristics of the 3 MC populations in trachea (colored as in a and compared with splenic MCp (grey shaded) and intraperitoneal MCs (back dotted line). (d) Forward scatter characteristics of tracheal, splenic and intraperitoneal MC populations as in c. Histograms in c and d show the mean value from 2-3 mice/group from one of 5 experiments. Bar graphs represent mean (± SEM) from 4 experiments with 5-8 mice per group *p < 0.05, **p < 0.01, ***p < 0.001.
Mentions: The eMMCs transiently present on D1 in the trachea have the highest β7 integrin expression of MCs in the trachea (Figure 5a, red line and bar). The mature MMCs have less β7 integrin expression (Figure 5a, blue line and bar) than eMMCs while their β1 expression is similar (Figure 5b). The granularity by SSC is greater in MMCs than the eMMCs, which is greater than splenic MCps (Figure 5c, blue line, red line, and grey shaded area, respectively). Nonetheless, the SSC of MMCs is still lower than that of the tracheal FcεRIhiKithi CMCs, which is modest compared to peritoneal MCs (Figure 5c, orange line and black dotted line, respectively). The FSC of CMCs is lower than that of the inducible trachea eMMC and MMC populations, with the eMMCs being comparable to peritoneal MCs (Figure 5d).

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