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Research upregulation of CD23 (FcepsilonRII) expression in human airway smooth muscle cells (huASMC) in response to IL-4, GM-CSF, and IL-4/GM-CSF.

Belleau JT, Gandhi RK, McPherson HM, Lew DB - Clin Mol Allergy (2005)

Bottom Line: The protein content of IL-4/GMCSF stimulated cells was significantly elevated.Stimulation with IL-4 resulted in the phosphorylation of STAT-6 and an increase in the expression of the IL-2Rgammac.Upregulation of CD23 by IL-4/GM-CSF results in phenotypic changes in huASMC that could play a role in cell migration or a change in the synthetic function of the cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Children's Foundation Research Center at the Le Bonheur Children's Medical Center, University of Tennessee Health Science Center, 50 North Dunlap Street, Rm401, WPT, Memphis, TN 38103, USA. jcbelleau@yahoo.com

ABSTRACT

Background: Airway smooth muscle cells play a key role in remodeling that contributes to airway hyperreactivity. Airway smooth muscle remodeling includes hypertrophy and hyperplasia. It has been previously shown that the expression of CD23 on ASMC in rabbits can be induced by the IgE component of the atopic serum. We examined if other components of atopic serum are capable of inducing CD23 expression independent of IgE.

Methods: Serum starved huASMC were stimulated with either IL-4, GM-CSF, IL-13, IL-5, PGD2, LTD4, tryptase or a combination of IL-4, IL-5, IL-13 each with GM-CSF for a period of 24 h. CD23 expression was analyzed by flow cytometry, western blot, and indirect immunofluorescence.

Results: The CD23 protein expression was upregulated in huASMC in response to IL-4, GM-CSF, and IL-4/GM-CSF. The percentage of cells with increased fluorescence intensity above the control was 25.1 +/- 4.2% (IL-4), 15.6 +/- 2.7% (GM-CSF) and 32.9 +/- 13.9% (IL-4/GMCSF combination)(n = 3). The protein content of IL-4/GMCSF stimulated cells was significantly elevated. Expression of CD23 in response to IL-4, GM-CSF, IL-4/GM-CSF was accompanied by changes in cell morphology including depolymerization of isoactin fibers, cell spreading, and membrane ruffling. Western blot revealed abundant expression of the IL-4Ralpha and a low level expression of IL-2Rgammac in huASMC. Stimulation with IL-4 resulted in the phosphorylation of STAT-6 and an increase in the expression of the IL-2Rgammac.

Conclusion: CD23 on huASMC is upregulated by IL-4, GM-CSF, and IL-4/GM-CSF. The expression of CD23 is accompanied by an increase in cell volume and an increase in protein content per cell, suggesting hypertrophy. Upregulation of CD23 by IL-4/GM-CSF results in phenotypic changes in huASMC that could play a role in cell migration or a change in the synthetic function of the cells. Upregulation of CD23 in huASMC by IL-4 and GM-CSF can contribute to changes in huASMC and may provide an avenue for new therapeutic options in asthma targeting ASMC.

No MeSH data available.


Related in: MedlinePlus

IL-4Rα and IL-2Rγc Expression in huASMC. A: IL-4Rα and IL-2Rγc expression in huASMC at baseline. Unstarved huASMC lysates were subjected to western blot analysis for IL-4Rα and IL-2Rγc using polyclonal anti-IL-4Rα (1:500 dilution, Santa Cruz), or monoclonal anti-IL-2Rγc (1:125 dilution, R&D Systems, Inc.). The nitrocellulose membranes were incubated with a 1:1,000 dilution of anti-rabbit or anti-mouse horseradish peroxidase linked whole antibody (Amersham). The immunoreactive protein bands were detected by ECL (Amersham). IL-2Rγc is minimally expressed in huASMC while IL-4Rα is expressed abundantly in huASMC.
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Figure 5: IL-4Rα and IL-2Rγc Expression in huASMC. A: IL-4Rα and IL-2Rγc expression in huASMC at baseline. Unstarved huASMC lysates were subjected to western blot analysis for IL-4Rα and IL-2Rγc using polyclonal anti-IL-4Rα (1:500 dilution, Santa Cruz), or monoclonal anti-IL-2Rγc (1:125 dilution, R&D Systems, Inc.). The nitrocellulose membranes were incubated with a 1:1,000 dilution of anti-rabbit or anti-mouse horseradish peroxidase linked whole antibody (Amersham). The immunoreactive protein bands were detected by ECL (Amersham). IL-2Rγc is minimally expressed in huASMC while IL-4Rα is expressed abundantly in huASMC.

Mentions: Western blot analysis of IL-4Rα and IL-2Rγc in huASMC lysates showed the presence of these receptor components on huASMC. Figure 5 shows abundant expression of IL-4Rα and a low level expression of IL-2Rγc protein on huASMC. After stimulation of huASMC with IL-4 (0.4 nM) for 24 h, a two fold increase in γc expression was observed compared to the BSA vehicle control (Figure 6).


Research upregulation of CD23 (FcepsilonRII) expression in human airway smooth muscle cells (huASMC) in response to IL-4, GM-CSF, and IL-4/GM-CSF.

Belleau JT, Gandhi RK, McPherson HM, Lew DB - Clin Mol Allergy (2005)

IL-4Rα and IL-2Rγc Expression in huASMC. A: IL-4Rα and IL-2Rγc expression in huASMC at baseline. Unstarved huASMC lysates were subjected to western blot analysis for IL-4Rα and IL-2Rγc using polyclonal anti-IL-4Rα (1:500 dilution, Santa Cruz), or monoclonal anti-IL-2Rγc (1:125 dilution, R&D Systems, Inc.). The nitrocellulose membranes were incubated with a 1:1,000 dilution of anti-rabbit or anti-mouse horseradish peroxidase linked whole antibody (Amersham). The immunoreactive protein bands were detected by ECL (Amersham). IL-2Rγc is minimally expressed in huASMC while IL-4Rα is expressed abundantly in huASMC.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1173127&req=5

Figure 5: IL-4Rα and IL-2Rγc Expression in huASMC. A: IL-4Rα and IL-2Rγc expression in huASMC at baseline. Unstarved huASMC lysates were subjected to western blot analysis for IL-4Rα and IL-2Rγc using polyclonal anti-IL-4Rα (1:500 dilution, Santa Cruz), or monoclonal anti-IL-2Rγc (1:125 dilution, R&D Systems, Inc.). The nitrocellulose membranes were incubated with a 1:1,000 dilution of anti-rabbit or anti-mouse horseradish peroxidase linked whole antibody (Amersham). The immunoreactive protein bands were detected by ECL (Amersham). IL-2Rγc is minimally expressed in huASMC while IL-4Rα is expressed abundantly in huASMC.
Mentions: Western blot analysis of IL-4Rα and IL-2Rγc in huASMC lysates showed the presence of these receptor components on huASMC. Figure 5 shows abundant expression of IL-4Rα and a low level expression of IL-2Rγc protein on huASMC. After stimulation of huASMC with IL-4 (0.4 nM) for 24 h, a two fold increase in γc expression was observed compared to the BSA vehicle control (Figure 6).

Bottom Line: The protein content of IL-4/GMCSF stimulated cells was significantly elevated.Stimulation with IL-4 resulted in the phosphorylation of STAT-6 and an increase in the expression of the IL-2Rgammac.Upregulation of CD23 by IL-4/GM-CSF results in phenotypic changes in huASMC that could play a role in cell migration or a change in the synthetic function of the cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Children's Foundation Research Center at the Le Bonheur Children's Medical Center, University of Tennessee Health Science Center, 50 North Dunlap Street, Rm401, WPT, Memphis, TN 38103, USA. jcbelleau@yahoo.com

ABSTRACT

Background: Airway smooth muscle cells play a key role in remodeling that contributes to airway hyperreactivity. Airway smooth muscle remodeling includes hypertrophy and hyperplasia. It has been previously shown that the expression of CD23 on ASMC in rabbits can be induced by the IgE component of the atopic serum. We examined if other components of atopic serum are capable of inducing CD23 expression independent of IgE.

Methods: Serum starved huASMC were stimulated with either IL-4, GM-CSF, IL-13, IL-5, PGD2, LTD4, tryptase or a combination of IL-4, IL-5, IL-13 each with GM-CSF for a period of 24 h. CD23 expression was analyzed by flow cytometry, western blot, and indirect immunofluorescence.

Results: The CD23 protein expression was upregulated in huASMC in response to IL-4, GM-CSF, and IL-4/GM-CSF. The percentage of cells with increased fluorescence intensity above the control was 25.1 +/- 4.2% (IL-4), 15.6 +/- 2.7% (GM-CSF) and 32.9 +/- 13.9% (IL-4/GMCSF combination)(n = 3). The protein content of IL-4/GMCSF stimulated cells was significantly elevated. Expression of CD23 in response to IL-4, GM-CSF, IL-4/GM-CSF was accompanied by changes in cell morphology including depolymerization of isoactin fibers, cell spreading, and membrane ruffling. Western blot revealed abundant expression of the IL-4Ralpha and a low level expression of IL-2Rgammac in huASMC. Stimulation with IL-4 resulted in the phosphorylation of STAT-6 and an increase in the expression of the IL-2Rgammac.

Conclusion: CD23 on huASMC is upregulated by IL-4, GM-CSF, and IL-4/GM-CSF. The expression of CD23 is accompanied by an increase in cell volume and an increase in protein content per cell, suggesting hypertrophy. Upregulation of CD23 by IL-4/GM-CSF results in phenotypic changes in huASMC that could play a role in cell migration or a change in the synthetic function of the cells. Upregulation of CD23 in huASMC by IL-4 and GM-CSF can contribute to changes in huASMC and may provide an avenue for new therapeutic options in asthma targeting ASMC.

No MeSH data available.


Related in: MedlinePlus