Expression of cytokine and chemokine mRNA and secretion of tumor necrosis factor-α by gallbladder epithelial cells: Response to bacterial lipopolysaccharides

Background

In addition to immune cells, many other cell types are known to produce cytokines. Cultured normal mouse gallbladder epithelial cells, used as a model system for gallbladder epithelium, were examined for their ability to express the mRNA of various cytokines and chemokines in response to bacterial lipopolysaccharide. The synthesis and secretion of the tumor necrosis factor-α (TNF-α) protein by these cells was also measured.

Results

Untreated mouse gallbladder cells expressed mRNA for TNF-α, RANTES, and macrophage inflammatory protein-2 (MIP-2). Upon treatment with lipopolysaccharide, these cells now produced mRNA for Interleukin-1β (IL-1β), IL-6, monocyte chemoattractant protein-1 (MCP-1), and showed increased expression of TNF-α and MIP-2 mRNA. Untreated mouse gallbladder cells did not synthesize TNF-α protein; however, they did synthesize and secrete TNF-α upon treatment with lipopolysaccharide.

Methods

Cells were treated with lipopolysaccharides from 3 strains of bacteria. Qualitative and semi-quantitative RT-PCR, using cytokine or chemokine-specific primers, was used to measure mRNA levels of TNFα, IL-1β, IL-6, IL-10, KC, RANTES, MCP-1, and MIP-2. TNF-α protein was measured by immunoassays.

Conclusion

This research demonstrates that gallbladder epithelial cells in response to lipopolysaccharide exposure can alter their cytokine and chemokine RNA expression pattern and can synthesize and secrete TNFα protein. This suggests a mechanism whereby gallbladder epithelial cells in vivo may mediate gallbladder secretory function, inflammation and diseases in an autocrine/paracrine fashion by producing and secreting cytokines and/or chemokines during sepsis.     

Response to the letter to the editor for “Obstructive Sleep Apnea and Risk of COVID-19 Infection,Hospitalization and Respiratory Failure”

Sleep and Breathing -     

Does the Number of Previous Vaginal Deliveries Affect Overactive Bladder Symptoms and their Response to Treatment?

Objectives: To explore associations between the number of vaginal deliveries (primary aim) or gender (secon‐ dary aim) and overactive bladder (OAB) symptoms as well as their response to treatment with a muscarinic antagonist. Methods: Preplanned secondary analysis of an observational study of solifenacin in OAB patients. Episode frequencies of OAB symptoms, pad use and scores on OAB rating scales were documented in 4450 patients before and after a 12–14 week treatment period with solifenacin 5 or 10 mg. Results: Women without, and with one, two or more than two vaginal deliveries and men were similar in their baseline characteristics. All groups also exhibited rather similar reductions in symptoms and improvements in rating scales upon treatment. Conclusion: These data indicate that solifenacin, and perhaps other muscarinic receptor antagonists, are similarly suitable for the treatment of OAB symptoms in both genders, irrespective of previous vaginal deliveries.     

Response to “Effect of APAP and heated humidification with a heated breathing tube on adherence,quality of life,and nasopharyngeal complaints”

Sleep and Breathing -     

Response to ‘Ultrasound characterization of cutaneous ulcers in systemic sclerosis’

Clinical Rheumatology -     

Predictive Factors of Response of Perianal Crohn’s Disease to Azathioprine or 6-Mercaptopurine

PURPOSE: This study was designed to evaluate the predictive factors of response of perianal Crohns disease to azathioprine or 6-mercaptopurine. METHODS: Ninety-four patients (65 females; mean age, 31 years) with active perianal Crohns disease were treated with azathioprine or 6-mercaptopurine for more than 6 (median, 27) months (median azathioprine dose, 2 mg/kg/day). The evolution of perianal lesions during azathioprine or 6-mercaptopurine therapy was analyzed retrospectively. Patients who had a clear anatomic improvement (fistula closure, fissure healing, stricture dilatation) and who did not develop any perianal complications requiring an antibiotic course or surgical intervention were considered responders regarding their perianal disease. RESULTS: Three years after inclusion, the cumulative probabilities of remaining free of perianal complication and achieving a clear anatomic improvement were 0.47 (95 percent confidence interval, 0.36–0.58) and 0.4 (95 percent confidence interval, 0.29–0.53), respectively. On the whole, 27 patients (29 percent) were responders to azathioprine or 6-mercaptopurine therapy. The absence of fistula, duration of perianal disease shorter than 22 months, and aged 40 years or older at inclusion were three independent factors associated with response to azathioprine or 6-mercaptopurine therapy. There was no correlation between the response of perianal lesions and the achievement of intestinal remission with azathioprine or 6-mercaptopurine. CONCLUSION: One-third of patients with perianal lesions of Crohns disease demonstrated a clear improvement during azathioprine or 6-mercaptopurine therapy. Patients aged 40 years or older with a recent perianal disease and without fistula were the best responders.     

Response to “Quantitative measures of tissue mechanics to detect hypermobile Ehlers Danlos syndrome and hypermobility syndrome disorders: a systematic review”

Clinical Rheumatology -     

Human Lung Fibroblast Response to NGF,IL-1β, and Dexamethsone

It has been shown that lung mast cells, eosinophils, and fibroblasts are receptive to the action of nerve growth factor (NGF) and that NGF is released in to the bloodstream of subjects affected by allergic inflammatory response. The role of NGF in lung inflammatory disorders is unclear because there is evidence suggesting that NGF can be involved in both proinflammatory and anti-inflammatory responses. Lung fibroblasts play a marked role in inflammation. In this study we investigated the effect of NGF, interleukin 1β (II-1β), and dexamethasone (DEX) on human lung fibroblasts in vitro. We found that II-1β, but not NGF, promotes fibroblasts’ survival and that NGF stimulates trkA receptor expression, down regulates TFG-α, and has no effect on TNF-β immunoreactivity. Moreover, DEX exerts different effects on NGF release by fibroblasts pre-exposed to II-1β. Our findings suggest that the NGF released by lung fibroblast during inflammation is not associated with the increase of proinflammatory factors such as TNF-α and II-1β.