Research Upregulation of CD23 FcεRII Expression in Human Airway Smooth Muscle Cells huASMC in Response to IL-4, GM-CSF, and IL-4-GM-CSFReport as inadecuate




Research Upregulation of CD23 FcεRII Expression in Human Airway Smooth Muscle Cells huASMC in Response to IL-4, GM-CSF, and IL-4-GM-CSF - Download this document for free, or read online. Document in PDF available to download.

Clinical and Molecular Allergy

, 3:6

First Online: 20 May 2005Received: 30 August 2004Accepted: 20 May 2005

Abstract

BackgroundAirway 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.

MethodsSerum 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.

ResultsThe 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 combinationn = 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-4Rα and a low level expression of IL-2Rγc in huASMC. Stimulation with IL-4 resulted in the phosphorylation of STAT-6 and an increase in the expression of the IL-2Rγc.

ConclusionCD23 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.

List of abbreviationshuASMCHuman airway smooth muscle cells

FceRIHigh-affinity receptor for IgE

FceRII CD23Low-affinity receptor for IgE

AHRAirway hyperreactivity

PGD2Prostaglandin D2

LTD4Leukotriene D4

PEPhycoerythrin

FBSFetal bovine serum

BSABovine serum albumin

PBSPhosphate bufferred saline

BCABicinchoninic acid

FITCFluorescein Isothiocyanate

TRITCTetramethyl Rhodamine Iso-Thiocyanate

RIPARadio-immunoprecipitation assay

ECLEnhanced chemiluminescence light

RαReceptor alpha

γcCommon gamma chain

FACSFluorescent Activated Cell Sorter

STATSignal Transducer Activator of Transcription

TH2T helper cell type 2

ERKExtracellular regulated kinases

Electronic supplementary materialThe online version of this article doi:10.1186-1476-7961-3-6 contains supplementary material, which is available to authorized users.

Joseph T Belleau, Radha K Gandhi contributed equally to this work.

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Author: Joseph T Belleau - Radha K Gandhi - Holly M McPherson - D Betty Lew

Source: https://link.springer.com/







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