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Airway smooth muscle proliferation and survival is not modulated by mast cells.

Kaur D, Hollins F, Saunders R, Woodman L, Sutcliffe A, Cruse G, Bradding P, Brightling C - Clin. Exp. Allergy (2009)

Bottom Line: Mast cell activation was confirmed by the measurement of histamine release.Using a number of techniques, we found that ASM proliferation and survival was not significantly different between cells derived from subjects with or without asthma.Our findings do not support a role for increased airway smooth proliferation and survival as the major mechanism driving ASM hyperplasia in asthma.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection, Inflammation and Immunity, Institute for Lung Health, University of Leicester, Leicester, UK.

ABSTRACT

Background: Airway smooth muscle (ASM) hyperplasia and mast cell localization within the ASM bundle are important features of asthma. The cause of this increased ASM mass is uncertain and whether it is a consequence of ASM-mast cell interactions is unknown.

Objective: We sought to investigate ASM proliferation and survival in asthma and the effects of co-culture with mast cells.

Methods: Primary ASM cultures were derived from 11 subjects with asthma and 12 non-asthmatic controls. ASM cells were cultured for up to 10 days in the presence or absence of serum either alone or in co-culture with the human mast cell line-1, unstimulated human lung mast cells (HLMC) or IgE/anti-IgE-activated HLMC. Proliferation was assessed by cell counts, CFSE assay and thymidine incorporation. Apoptosis and necrosis were analysed by Annexin V/propidium iodide staining using flow cytometry and by assessment of nuclear morphology using immunofluorescence. Mast cell activation was confirmed by the measurement of histamine release.

Results: Using a number of techniques, we found that ASM proliferation and survival was not significantly different between cells derived from subjects with or without asthma. Co-culture with mast cells did not affect the rate of proliferation or survival of ASM cells.

Conclusion: Our findings do not support a role for increased airway smooth proliferation and survival as the major mechanism driving ASM hyperplasia in asthma.

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Related in: MedlinePlus

Airway smooth muscle (ASM) cells (n=4) were co-cultured with un-activated human lung mast cells (HLMC, n=4) alone and activated HLMC (n=4, IgE/Anti-IgE) over 10 days in ITS media. Histamine concentrations were measured in culture supernatants and corrected for mast cell number. Constitutive histamine release was demonstrated with un-activated HLMC co-culture, which was augmented in IgE/anti-IgE activated HLMC over 1 and 3 days. Data presented as mean±SEM; P-values are as shown.
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fig03: Airway smooth muscle (ASM) cells (n=4) were co-cultured with un-activated human lung mast cells (HLMC, n=4) alone and activated HLMC (n=4, IgE/Anti-IgE) over 10 days in ITS media. Histamine concentrations were measured in culture supernatants and corrected for mast cell number. Constitutive histamine release was demonstrated with un-activated HLMC co-culture, which was augmented in IgE/anti-IgE activated HLMC over 1 and 3 days. Data presented as mean±SEM; P-values are as shown.

Mentions: HLMC alone (n=4) released significantly more histamine with IgE/anti-IgE activation at day 0 (918±101 ng/106 cells) when compared with un-activated HLMC (495±85 ng/106 cells; P=0.001). In co-culture, histamine was readily measured at all time-points and increased over time with concentrations at day 7 (P=0.03) and 10 (P=0.02) significantly higher than day 1 (Fig. 3). Histamine release by HLMC co-cultured with ASM cells and IgE/anti-IgE activated was increased compared with unstimulated co-cultured HLMC after 1 and 3 days (Fig. 3).


Airway smooth muscle proliferation and survival is not modulated by mast cells.

Kaur D, Hollins F, Saunders R, Woodman L, Sutcliffe A, Cruse G, Bradding P, Brightling C - Clin. Exp. Allergy (2009)

Airway smooth muscle (ASM) cells (n=4) were co-cultured with un-activated human lung mast cells (HLMC, n=4) alone and activated HLMC (n=4, IgE/Anti-IgE) over 10 days in ITS media. Histamine concentrations were measured in culture supernatants and corrected for mast cell number. Constitutive histamine release was demonstrated with un-activated HLMC co-culture, which was augmented in IgE/anti-IgE activated HLMC over 1 and 3 days. Data presented as mean±SEM; P-values are as shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2821816&req=5

fig03: Airway smooth muscle (ASM) cells (n=4) were co-cultured with un-activated human lung mast cells (HLMC, n=4) alone and activated HLMC (n=4, IgE/Anti-IgE) over 10 days in ITS media. Histamine concentrations were measured in culture supernatants and corrected for mast cell number. Constitutive histamine release was demonstrated with un-activated HLMC co-culture, which was augmented in IgE/anti-IgE activated HLMC over 1 and 3 days. Data presented as mean±SEM; P-values are as shown.
Mentions: HLMC alone (n=4) released significantly more histamine with IgE/anti-IgE activation at day 0 (918±101 ng/106 cells) when compared with un-activated HLMC (495±85 ng/106 cells; P=0.001). In co-culture, histamine was readily measured at all time-points and increased over time with concentrations at day 7 (P=0.03) and 10 (P=0.02) significantly higher than day 1 (Fig. 3). Histamine release by HLMC co-cultured with ASM cells and IgE/anti-IgE activated was increased compared with unstimulated co-cultured HLMC after 1 and 3 days (Fig. 3).

Bottom Line: Mast cell activation was confirmed by the measurement of histamine release.Using a number of techniques, we found that ASM proliferation and survival was not significantly different between cells derived from subjects with or without asthma.Our findings do not support a role for increased airway smooth proliferation and survival as the major mechanism driving ASM hyperplasia in asthma.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection, Inflammation and Immunity, Institute for Lung Health, University of Leicester, Leicester, UK.

ABSTRACT

Background: Airway smooth muscle (ASM) hyperplasia and mast cell localization within the ASM bundle are important features of asthma. The cause of this increased ASM mass is uncertain and whether it is a consequence of ASM-mast cell interactions is unknown.

Objective: We sought to investigate ASM proliferation and survival in asthma and the effects of co-culture with mast cells.

Methods: Primary ASM cultures were derived from 11 subjects with asthma and 12 non-asthmatic controls. ASM cells were cultured for up to 10 days in the presence or absence of serum either alone or in co-culture with the human mast cell line-1, unstimulated human lung mast cells (HLMC) or IgE/anti-IgE-activated HLMC. Proliferation was assessed by cell counts, CFSE assay and thymidine incorporation. Apoptosis and necrosis were analysed by Annexin V/propidium iodide staining using flow cytometry and by assessment of nuclear morphology using immunofluorescence. Mast cell activation was confirmed by the measurement of histamine release.

Results: Using a number of techniques, we found that ASM proliferation and survival was not significantly different between cells derived from subjects with or without asthma. Co-culture with mast cells did not affect the rate of proliferation or survival of ASM cells.

Conclusion: Our findings do not support a role for increased airway smooth proliferation and survival as the major mechanism driving ASM hyperplasia in asthma.

Show MeSH
Related in: MedlinePlus