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Published on: April 29, 2015
Monocyte derived macrophages from CF pigs exhibit increased inflammatory responses at birth
Lily Paemka1, Brian N McCullagh1, Mahmoud H Abou Alaiwa2
1Department of Pediatrics, Pappajohn Biomedical Institute, Carver College of Medicine, University of Iowa, Iowa City, IA, USA.
Background:
We sought to address whether CF macrophages have a primary functional defect as a consequence of CFTR loss and thus contribute to the onset of infection and inflammation observed in CF lung disease.
Methods:
Monocyte derived macrophages (MDMs) were prepared from newborn CF and non-CF pigs. CFTR mRNA expression was quantified by rtPCR and anion channel function was determined using whole cell patch clamp analysis. IL8 and TNFα release from MDMs in response to lipopolysaccharide stimulation was measured by ELISA.
Results:
CFTR was expressed in MDMs by Q-rtPCR at a lower level than in epithelial cells. MDMs exhibited functional CFTR current at the cell membrane and this current was absent in CF MDMs. CF MDMs demonstrated an exaggerated response to lipopolysaccharide stimulation.
Conclusions:
In the absence of CFTR function, macrophages from newborn CF pigs exhibit an increased inflammatory response to a lipopolysaccharide challenge. This may contribute to the onset and progression of CF lung disease.
Insights
Macrophages from newborn cystic fibrosis (CF) pigs show a heightened inflammatory response due to CFTR loss. This defect may drive the lung disease progression in CF patients.
Area of Science:
- Immunology
- Cell Biology
- Genetics
Background:
- Cystic fibrosis (CF) lung disease is characterized by chronic infection and inflammation.
- The role of macrophages in the early pathogenesis of CF lung disease remains unclear.
Purpose of the Study:
- To investigate whether cystic fibrosis transmembrane conductance regulator (CFTR) loss in macrophages causes primary functional defects.
- To determine if these defects contribute to the onset of infection and inflammation in CF lung disease.
Main Methods:
- Monocyte-derived macrophages (MDMs) were isolated from newborn CF and non-CF pigs.
- CFTR mRNA expression was quantified using quantitative reverse transcription polymerase chain reaction (Q-rtPCR).
- Anion channel function was assessed via whole-cell patch-clamp analysis, and cytokine release (IL-8, TNF-α) was measured by ELISA following lipopolysaccharide stimulation.
Main Results:
- CFTR mRNA was detected at lower levels in MDMs compared to epithelial cells.
- Functional CFTR currents were present in non-CF MDMs but absent in CF MDMs.
- CF MDMs exhibited an exaggerated inflammatory response to lipopolysaccharide stimulation.
Conclusions:
- Macrophages lacking CFTR function display an amplified inflammatory response to lipopolysaccharide.
- This heightened inflammatory response in CF macrophages may contribute to the initiation and advancement of CF lung disease.
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