Exposure to filtered air, cigarette smoke, and LTβR-Ig fusion protein as described in the paper. Briefly, Cigarette smoke (CS) was generated from 3R4F Research Cigarettes (Tobacco Research Institute, University of Kentucky, Lexington, KY), with the filters removed. Mice were whole body exposed to active 100% mainstream CS of 500 mg/m3 total particulate matter (TPM) for 50 min twice per day for 4 and 6 months (m) in a manner mimicking natural human smoking habits as previously described. The TPM level was monitored via gravimetric analysis of quartz fiber filters prior and after sampling air from the exposure chamber and measuring the total air volume. CO concentrations in the exposure chamber were constantly monitored by using a GCO 100 CO Meter (Greisinger Electronic, Regenstauf, Germany) and reached values of 288± 74 ppm. All mice tolerated CS-mediated CO concentrations without any sign of toxicity, with CO-Hb levels of 12.2 ± 2.4%. In two parallel experiments, mice were treated with an LTβR-Ig fusion protein (80 µg i.p., weekly) (muLTβR-muIgG1, ref #14811-122; and CHO 14.3 Lot #7195-30) or control-Ig (MOPC21, clone #113) for 2m, starting from 2m and 4m of CS exposure. Control mice were kept in a filtered air (FA) environment, but exposed to the same stress as CS-exposed animals. 24 h after the last CS exposure, mice were sacrificed.
Growth protocol
Animals housed under spf conditions
Extracted molecule
total RNA
Extraction protocol
RNeasy Mini Kit (Qiagen) including DNase treatment
Label
Biotin
Label protocol
Illumina TotalPrep RNA Amplification kit (Ambion)
Hybridization protocol
Standard Illumina hybridization protocol
Scan protocol
Standard Illumina scanning protocol
Description
Gene expression data from lung tissue from mice exposed to cigarette smoke for 6 months plus the LTβR-Ig fusion protein, biological replicate 3
Data processing
Data including background subtraction (GenomeStudioV2011.1 software, gene expression module version 1.9.0), removal of negative values by introducing an offset, quantile normalisation
