Gene expression profiling of Treg subsets based on ILT3 expression
Comparison of gene expression ex vivo isolated ILT3+ respectively ILT3- Treg cells Overall design: 10-16-week-old syngeneic male and female mice were euthanized via CO2 inhalation. Spleens and mesenteric lymph nodes were mechanically disrupted and filtered through a 40 µm cell strainer to remove cell debris. Erythrocytes of spleen suspension were removed through the use of Gey's lysis buffer and filtered again through a 40µM cell strainer. Live cells were counted microscopically by exclusion of dead cells with trypan blue staining. CD25+ CD4+ T cells were pre-enriched via magentic columns. ILT3+ respectively ILT3- CD4+ CD25+ T cells were sorted with a FACSAria III (BD Biosciences).