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Influence of the cystic fibrosis transmembrane conductance regulator on expression of lipid metabolism-related genes in dendritic cells.

Xu Y, Tertilt C, Krause A, Quadri LE, Crystal RG, Worgall S - Respir. Res. (2009)

Bottom Line: Gene expression analysis in DC generated from naive CF and WT mice revealed decreased expression of Caveolin-1 (Cav1), a membrane lipid raft protein, in the CF DC compared to WT DC.Following exposure to P. aeruginosa, expression of 3beta-hydroxysterol-Delta7 reductase (Dhcr7) and stearoyl-CoA desaturase 2 (Scd2), two enzymes involved in the lipid metabolism that are also regulated by SREBP, was less decreased in the CF DC compared to WT DC.These results suggest that CFTR dysfunction in DC affects factors involved in membrane structure and lipid-metabolism, which may contribute to the abnormal inflammatory and immune response characteristic of CF.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Weill Cornell Medical College, New York, USA. yax2002@med.cornell.edu

ABSTRACT

Background: Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Infections of the respiratory tract are a hallmark in CF. The host immune responses in CF are not adequate to eradicate pathogens, such as P. aeruginosa. Dendritic cells (DC) are crucial in initiation and regulation of immune responses. Changes in DC function could contribute to abnormal immune responses on multiple levels. The role of DC in CF lung disease remains unknown.

Methods: This study investigated the expression of CFTR gene in bone marrow-derived DC. We compared the differentiation and maturation profile of DC from CF and wild type (WT) mice. We analyzed the gene expression levels in DC from naive CF and WT mice or following P. aeruginosa infection.

Results: CFTR is expressed in DC with lower level compared to lung tissue. DC from CF mice showed a delayed in the early phase of differentiation. Gene expression analysis in DC generated from naive CF and WT mice revealed decreased expression of Caveolin-1 (Cav1), a membrane lipid raft protein, in the CF DC compared to WT DC. Consistently, protein and activity levels of the sterol regulatory element binding protein (SREBP), a negative regulator of Cav1 expression, were increased in CF DC. Following exposure to P. aeruginosa, expression of 3beta-hydroxysterol-Delta7 reductase (Dhcr7) and stearoyl-CoA desaturase 2 (Scd2), two enzymes involved in the lipid metabolism that are also regulated by SREBP, was less decreased in the CF DC compared to WT DC.

Conclusion: These results suggest that CFTR dysfunction in DC affects factors involved in membrane structure and lipid-metabolism, which may contribute to the abnormal inflammatory and immune response characteristic of CF.

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Related in: MedlinePlus

CFTR expression in bone marrow derived dendritic cells (DC). RNA and protein were extracted from wild type (WT) mouse lung and DC. CFTR expression was measured by real-time RT-PCR and Western analysis. A. Real-time RT-PCR. WT mouse lung tissue was used as a positive control and calibration. The y-axis represents CFTR cDNA transcription level in terms of relative quantity value (RQ). B. Western analysis of CFTR protein in DC compared to the WT lung tissue. C. Quantification of CFTR protein by image intensity analysis. Images were scanned and analyzed by software Image J normalized to GAPDH loading control. Shown is the mean ± SEM of three pairs of independent samples. **denotes p < 0.01.
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Figure 1: CFTR expression in bone marrow derived dendritic cells (DC). RNA and protein were extracted from wild type (WT) mouse lung and DC. CFTR expression was measured by real-time RT-PCR and Western analysis. A. Real-time RT-PCR. WT mouse lung tissue was used as a positive control and calibration. The y-axis represents CFTR cDNA transcription level in terms of relative quantity value (RQ). B. Western analysis of CFTR protein in DC compared to the WT lung tissue. C. Quantification of CFTR protein by image intensity analysis. Images were scanned and analyzed by software Image J normalized to GAPDH loading control. Shown is the mean ± SEM of three pairs of independent samples. **denotes p < 0.01.

Mentions: First we evaluated the level of CFTR expression in DC compared to lung tissue known for high expression of CFTR. CFTR mRNA was detected in DC and whole lung by real-time RT-PCR (Figure 1A). The CFTR mRNA levels were 212-fold lower in the DC compared to the whole lung (p < 0.01). Likewise, CFTR protein was detected by Western analysis (Figure 1B); the expression level in DC was 11-fold lower compared to lung (p < 0.01, Figure 1C).


Influence of the cystic fibrosis transmembrane conductance regulator on expression of lipid metabolism-related genes in dendritic cells.

Xu Y, Tertilt C, Krause A, Quadri LE, Crystal RG, Worgall S - Respir. Res. (2009)

CFTR expression in bone marrow derived dendritic cells (DC). RNA and protein were extracted from wild type (WT) mouse lung and DC. CFTR expression was measured by real-time RT-PCR and Western analysis. A. Real-time RT-PCR. WT mouse lung tissue was used as a positive control and calibration. The y-axis represents CFTR cDNA transcription level in terms of relative quantity value (RQ). B. Western analysis of CFTR protein in DC compared to the WT lung tissue. C. Quantification of CFTR protein by image intensity analysis. Images were scanned and analyzed by software Image J normalized to GAPDH loading control. Shown is the mean ± SEM of three pairs of independent samples. **denotes p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2683168&req=5

Figure 1: CFTR expression in bone marrow derived dendritic cells (DC). RNA and protein were extracted from wild type (WT) mouse lung and DC. CFTR expression was measured by real-time RT-PCR and Western analysis. A. Real-time RT-PCR. WT mouse lung tissue was used as a positive control and calibration. The y-axis represents CFTR cDNA transcription level in terms of relative quantity value (RQ). B. Western analysis of CFTR protein in DC compared to the WT lung tissue. C. Quantification of CFTR protein by image intensity analysis. Images were scanned and analyzed by software Image J normalized to GAPDH loading control. Shown is the mean ± SEM of three pairs of independent samples. **denotes p < 0.01.
Mentions: First we evaluated the level of CFTR expression in DC compared to lung tissue known for high expression of CFTR. CFTR mRNA was detected in DC and whole lung by real-time RT-PCR (Figure 1A). The CFTR mRNA levels were 212-fold lower in the DC compared to the whole lung (p < 0.01). Likewise, CFTR protein was detected by Western analysis (Figure 1B); the expression level in DC was 11-fold lower compared to lung (p < 0.01, Figure 1C).

Bottom Line: Gene expression analysis in DC generated from naive CF and WT mice revealed decreased expression of Caveolin-1 (Cav1), a membrane lipid raft protein, in the CF DC compared to WT DC.Following exposure to P. aeruginosa, expression of 3beta-hydroxysterol-Delta7 reductase (Dhcr7) and stearoyl-CoA desaturase 2 (Scd2), two enzymes involved in the lipid metabolism that are also regulated by SREBP, was less decreased in the CF DC compared to WT DC.These results suggest that CFTR dysfunction in DC affects factors involved in membrane structure and lipid-metabolism, which may contribute to the abnormal inflammatory and immune response characteristic of CF.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Weill Cornell Medical College, New York, USA. yax2002@med.cornell.edu

ABSTRACT

Background: Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Infections of the respiratory tract are a hallmark in CF. The host immune responses in CF are not adequate to eradicate pathogens, such as P. aeruginosa. Dendritic cells (DC) are crucial in initiation and regulation of immune responses. Changes in DC function could contribute to abnormal immune responses on multiple levels. The role of DC in CF lung disease remains unknown.

Methods: This study investigated the expression of CFTR gene in bone marrow-derived DC. We compared the differentiation and maturation profile of DC from CF and wild type (WT) mice. We analyzed the gene expression levels in DC from naive CF and WT mice or following P. aeruginosa infection.

Results: CFTR is expressed in DC with lower level compared to lung tissue. DC from CF mice showed a delayed in the early phase of differentiation. Gene expression analysis in DC generated from naive CF and WT mice revealed decreased expression of Caveolin-1 (Cav1), a membrane lipid raft protein, in the CF DC compared to WT DC. Consistently, protein and activity levels of the sterol regulatory element binding protein (SREBP), a negative regulator of Cav1 expression, were increased in CF DC. Following exposure to P. aeruginosa, expression of 3beta-hydroxysterol-Delta7 reductase (Dhcr7) and stearoyl-CoA desaturase 2 (Scd2), two enzymes involved in the lipid metabolism that are also regulated by SREBP, was less decreased in the CF DC compared to WT DC.

Conclusion: These results suggest that CFTR dysfunction in DC affects factors involved in membrane structure and lipid-metabolism, which may contribute to the abnormal inflammatory and immune response characteristic of CF.

Show MeSH
Related in: MedlinePlus