Mice, Cytokines, Antibodies, and Other Reagents
BALB/c DO11.10 mice transgenic for OVA-specific TCR WT or crossed back with Rag1-, IL-4-, IFN-γ-, STAT4-, and STAT6-deficient mice were used as a source of antigen-specific T cells (Murphy et al., 1990; Ouyang et al., 1998; Shoemaker et al., 2006) and were bred and maintained under SPF conditions at the NIMR, London, Home Office, UK, Animals (Scientific Procedures) Act 1986 or at the Washington University School of Medicine. Female mice were used at 8–12 weeks old, and animal protocols were approved according to the Animals (Scientific Procedures) Act 1986, Home Office, UK. Reagents, including antibodies for T cell and DC preparation, purification and culture, media, cytokines, and cytokine mAbs have been described (Hosken et al., 1995; Shoemaker et al., 2006; Veldhoen et al., 2009; Veldhoen et al., 2006). LPS (S. minnesota) was from Alexis, chicken ovalbumin protein (OVA protein) was from from Sigma-Aldrich, and ovalbumin peptide323-339 (OVA) (endotoxin-free) was from Biosynthesis. U0126 was from BioMol International. PD184352 (MEK inhibitors), SB203580 (p38 inhibitor), and CT99021 (GSK3β inhibitor) were kind gifts from P. Cohen and N. Shpiro, University of Dundee, UK.