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Published on: November 16, 2016
Several gene programs are induced in ciprofloxacin-treated human lymphocytes as revealed by microarray analysis
Emily Eriksson1, Arne Forsgren, Kristian Riesbeck
1Department of Medical Microbiology, Lund University, Malmö University Hospital, Sweden.
Abstract:
Fluoroquinolones have immunomodulatory properties and interfere with cytokine production. The aim of this study was to characterize the extent of the superinduced mRNA levels in activated human lymphocytes incubated with ciprofloxacin (5 and 80 micro g/ml) using a cytokine gene expression microarray from R and D Systems (Abingdon, UK). Several gene transcripts (n=104) were up-regulated in cells treated with ciprofloxacin at 80 micro g/ml, whereas 98 transcripts were down-regulated out of 847 total genes included on the microarray. The increased mRNAs were distributed between major gene programs, including interleukins (36.5%), signal-transduction molecules (13.5%), adhesion molecules (10.6%), tumor necrosis factor and transforming growth factor-beta superfamilies (10.6%), cell-cycle regulators (9.6%), and apoptosis-related molecules (8.7%). To determine the specificity of the microarray, a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), which contained a panel of 12 different cytokine mRNAs, was used. Eleven out of the 12 gene transcripts were up-regulated in the specific RT-PCR, whereas only eight were found to be increased in the microarray. A microarray from Clontech (Hampshire, UK), containing 588 different genes, was also included. Results obtained with this broad-coverage expression array slightly differed compared with the other microarray. We conclude that the fluoroquinolone ciprofloxacin at high concentrations interferes with several gene programs, which is in accordance with a mammalian stress response. From a technical point of view, a discrepancy may exist between data obtained by different microarrays and more specific methods such as quantitative RT-PCR.
Insights
The antibiotic ciprofloxacin, at high concentrations, alters gene expression in human immune cells, impacting key biological pathways. This suggests a potential stress response and highlights differences in gene expression analysis methods.
Area of Science:
- Immunology
- Molecular Biology
- Pharmacology
Background:
- Fluoroquinolones possess immunomodulatory effects.
- These antibiotics can interfere with cytokine production.
- Understanding ciprofloxacin's impact on gene expression is crucial.
Purpose of the Study:
- To investigate the impact of ciprofloxacin on mRNA levels in activated human lymphocytes.
- To characterize gene expression changes using cytokine gene expression microarrays.
- To compare microarray data with quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).
Main Methods:
- Activated human lymphocytes were incubated with ciprofloxacin (5 and 80 µg/ml).
- Gene expression profiling was performed using cytokine gene expression microarrays (R and D Systems, Clontech).
- Quantitative RT-PCR was employed to validate microarray findings for specific cytokine mRNAs.
Main Results:
- Ciprofloxacin at 80 µg/ml significantly altered mRNA levels, up-regulating 104 and down-regulating 98 transcripts.
- Affected gene programs included interleukins, signal-transduction molecules, adhesion molecules, and apoptosis-related genes.
- Quantitative RT-PCR confirmed up-regulation for 11 out of 12 tested cytokine mRNAs, showing higher sensitivity than one microarray.
Conclusions:
- High concentrations of ciprofloxacin interfere with multiple gene programs in human lymphocytes.
- These alterations are consistent with a mammalian stress response.
- Discrepancies between different microarray platforms and RT-PCR highlight the importance of method selection for accurate gene expression analysis.
