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SAMPLES (18)

mace:id
Technology # Array version
Species
Organ
Tissue
Physiopathology
Type
Name

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User name	Arndt Benecke
Email	arndt@ihes.fr
Phone / Fax number	+33160926665 / +33160926609
Location	Institut des Hautes Etudes Scientifiques (Integrative Biology) - 35, route de Chartres - 91440 Bures sur Yvette, France

Scientific description

Acute pneumonia caused by Streptococcus pneumoniae is a major cause of child mortality. Antibodies are considered the main effectors of protection against invasive disease. To get new insights into the mechanisms involved in the protective immunity against S. pneumoniae acute pneumonia we established an experimental model of protection in mice and then conducted genome-wide transcriptional analysis of lungs from protected and non-protected animals. Intranasal inoculation of a sub-lethal dose of S. pneumoniae serotype 1 conferred protection against a subsequent challenge with a lethal dose of the same strain. Microarray-based analysis of whole lung obtained 24 h after lethal challenge showed only 149 genes differentially expressed between protected and non-protected animals. These included upregulation of an important number of genes related to immunity and defense in protected animals, particularly Il17a, Il17f, Il21 and Il22 as well as Ifng and several IFN-ï§-dependent genes. We found also a significant increase in IL-17A and IFN-ï§ proteins in the lung of protected animals. Time course analysis showed that gene expression was upregulated as soon as 8h after challenge in protected animals but was diminished or strongly delayed in the control group. Overall, these results show that a timely upregulated Il17a- and Ifng related signature is associated to the protective immunity against S. pneumoniae acute pneumonia.

Technical description

Total RNA was extracted using NucleoSpinÂ® RNA L (Macherey-Nagel) and was concentrated in YM30 Microcon centrifugal filter device (Millipore) by centrifugation at 14 000 g during 60 min at 4 ÂºC. RNA quality was evaluated with an Agilent 2100 Bioanalyser (Agilent Technologies). Poly(A) mRNA from 20 Âµg of total RNA was converted into digoxigenin labeled cDNA for hybridization to Mouse Genome Survey Microarrays (Applied Biosystems) using Chemiluminescent RT Labeling Kit for detection in an Applied Biosystems ABI1700 system.

Data analysis 
Microarray data were quality-controlled (8). Statistical analysis of the microarray data was carried out as described previously (32, 33, 47)

Ontology Enrichment Analysis
Combining GO, Kegg and PANTHER annotations, we assigned all probes present on the MGS Version 2.0 array to the PANTHER biological processes. We then calculated the relative representation of those probes detected as significantly regulated. P-values for over- and under-representation of biological processes were calculated using a binominal distribution.

Reference

Pubmed : http://www.ncbi.nlm.nih.gov/pubmed/22204818