The purity of CD4+CD25+ or CD4+CD25− cells was 80–90% as assessed by flow cytometry. Then, 5×104 aliquots of WT or lpr DC were cultured in triplicate with 2.5×105 CD8+ T cells enriched from LNC of sensitized mice obtained at day +5 post-sensitization in complete RPMI-1640 media at 37oC, 5% CO2 and 5×104 aliquots of CD4+CD25+ T cells or CD4+CD25− T cells purified

from LN of naïve mice were added to these cultures. After 72 h of culture, supernatants were collected and tested for IFN-γ using Quantikine Mouse IFN-γ Immunoassay Kit (R&D Systems, Minneapolis, MN). Statistical analysis to assess differences between experimental groups was performed using two-tailed Student’s t test. Differences were considered significant when p<0.05. Three mice per group were used in all in vivo experiments. For in vitro experiments, three triplicate

samples PLX4032 clinical trial were analyzed for each group. All experiments were repeated at least two times with similar results. The authors thank the staff of the Cleveland Clinic Biological Resources Unit for excellent animal care. This work was supported by National Institutes of Health Grant RO1 AI45888 (R.L.F.). Conflict of interest: The authors declare no financial or commercial conflict of interest. “
“Over the last Crizotinib price decade, significant technological breakthroughs have revolutionized human genomic research in the form of genome-wide association studies (GWASs). GWASs have identified thousands of statistically significant genetic variants associated with hundreds of human conditions including many with immunological aetiologies (e.g. multiple sclerosis, ankylosing spondylitis and rheumatoid arthritis). Unfortunately, most GWASs fail to identify clinically significant associations. Identifying biologically significant variants by GWAS

also presents a challenge. The GWAS is a phenotype-to-genotype approach. As a complementary/alternative approach to the GWAS, investigators have begun to exploit extensive electronic medical record systems to conduct a genotype-to-phenotype approach when studying human disease – specifically, the phenome-wide Pregnenolone association study (PheWAS). Although the PheWAS approach is in its infancy, this method has already demonstrated its capacity to rediscover important genetic associations related to immunological diseases/conditions. Furthermore, PheWAS has the advantage of identifying genetic variants with pleiotropic properties. This is particularly relevant for HLA variants. For example, PheWAS results have demonstrated that the HLA-DRB1 variant associated with multiple sclerosis may also be associated with erythematous conditions including rosacea. Likewise, PheWAS has demonstrated that the HLA-B genotype is not only associated with spondylopathies, uveitis, and variability in platelet count, but may also play an important role in other conditions, such as mastoiditis.