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Project context and summary :
The LabEx Milieu interieur (MI) project is a clinical study aiming to define the natural variability of the human immune response. Fibroblasts were prepared from skin biopsies of 300 of the 1000 healthy donors (30 men and 30 women in each of 5 decades from 20 to 69 years) by Genethon. Each primary line is annotated by metadata derived from the systematic genotypic-phenotypic analysis Labex-MI (serology, genomic analyzes, proteomics, transcriptome, microbiota, clinical data and epidemiological selection criteria). The objective is to characterize the innate immune response of fibroblasts from healthy donors. We have developed approaches to obtain standardized measures of the immune response and to identify interindividual variance (Chansard et al., 2021) on a subset of sixteen primary fibroblast lines.. We observed that cell responses to LPS stimulation were distributed between “low” and “high” responders, while inter-donor variability of the response to poly I: C was very low. Poly I: C and LPS activate pathways from TLR3 and TLR4 receptors, respectively. We set up a real-time quantitative PCR assay of the 16 cells in parallel using the BioMark automated PCR system (Fluidigm) for comparative quantification of mRNAs baseline expression in the donors’ fibroblasts to check if low responses to LPS were due to a different expression of TLR4 and other factors of the pathway. We will extend the analysis to stimulated cells by Poly I: C ,LPS and other agonists of the innate immune response and also in parallel to cells infected by Leptospira interrogans.Related team publications :
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