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Updated: Apr 27, 2026
Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins
Published on: June 14, 2016
Differential response of gel-forming mucins to pathogenic middle ear bacteria
Joseph E Kerschner1, Wenzhou Hong2, Pawjai Khampang2
1Division of Pediatric Otolaryngology, Children's Hospital of Wisconsin, 9000 W. Wisconsin Avenue, Milwaukee, Wisconsin 53226, USA; Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, Wisconsin 53226, USA.
Objective:
To assess the differential response of the secretory gel forming mucins (GFM) to the most common bacterial pathogens causing otitis media, Streptococcus pneumoniae (SP), nontypeable Haemophilus influenza (NTHi), and Moraxella catarrhalis (Mcat), in a culture model of human middle ear epithelium (HMEEC).
Methods:
In vitro cultured HMEEC was exposed to 5 μg/ml of bacterial whole cell lysate (WCL). RNA was extracted to generate cDNA. The expression levels of each of the targeted mucin transcripts, MUC2, MUC5AC, MUC5B and MUC19, were detected by quantitative PCR.
Results:
The submerged HMEEC exposed to NTHi-86028NP WCL demonstrated a significant increase of MUC2, MUC5AC and MUC5B as compared to the control non-treated cells while MUC19 transcript level remained unchanged. WCL of additional major OM pathogens significantly increase the transcription of these three mucin genes as well. A combination of NTHi and SP further synergistically induced MUC2 and MUC5AC gene expression however, not all NTHi strains synergized with SP in the induction. Addition of Mcat WCL to the synergized combination of NTHi and SP did not participate in the synergistic response of mucins.
Conclusion:
The specific pathogen combinations were important in determining the degree of synergistic effects to GFM expression. The current data are substantive in guiding future work to extend our understanding of OM pathogens and GFMs.
Insights
Bacterial pathogens causing otitis media differentially regulate gel-forming mucins (GFM) in human middle ear epithelium. Specific pathogen combinations, like Streptococcus pneumoniae and nontypeable Haemophilus influenza, synergistically increase MUC2 and MUC5AC expression.
Area of Science:
- Otorhinolaryngology
- Microbiology
- Molecular Biology
Background:
- Otitis media (OM) is a common middle ear infection.
- Secretory gel-forming mucins (GFM) play a crucial role in mucosal defense.
- Understanding pathogen-specific mucin responses is vital for OM pathogenesis research.
Purpose of the Study:
- To investigate the differential expression of MUC2, MUC5AC, MUC5B, and MUC19 in human middle ear epithelial cells (HMEEC) upon exposure to common OM pathogens.
- To assess the synergistic effects of combined bacterial pathogens on GFM expression.
Main Methods:
- Human middle ear epithelial cells (HMEEC) were cultured in vitro.
- Cells were exposed to whole cell lysates (WCL) of Streptococcus pneumoniae (SP), nontypeable Haemophilus influenza (NTHi), and Moraxella catarrhalis (Mcat).
- Quantitative PCR was used to measure the expression levels of MUC2, MUC5AC, MUC5B, and MUC19 transcripts.
Main Results:
- NTHi WCL significantly increased MUC2, MUC5AC, and MUC5B expression in HMEEC, while MUC19 remained unchanged.
- Other major OM pathogens also significantly upregulated these three mucin genes.
- Combinations of NTHi and SP synergistically induced MUC2 and MUC5AC expression, though this effect varied by NTHi strain.
- M. catarrhalis did not contribute to the synergistic mucin response when combined with NTHi and SP.
Conclusions:
- Pathogen-specific interactions significantly influence the synergistic induction of gel-forming mucins in middle ear infections.
- These findings provide a basis for further research into the complex interplay between OM pathogens and host mucin responses.
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