Research+3-20-12

Some groups use peptide pulsed splenocytes as the targets In Vivo NK Cell and CD8+ T Cell Cytotoxicity Screen http://mutagenetix.utsouthwestern.edu/protocol/protocol_rec.cfm?pid=8 It looks like it could be extremely useful to buy the "Anti-Mouse OVA257-264 (SIINFEKL) peptide bound to H-2Kb PE" http://www.ebioscience.com/mouse-mhc-class-i-antibody-pe-ebio25-d116.htm These people use EL4 cells (T lymphocyte lymphoma cells from C57BL/6N as targets) Cellular protein is the source of cross-priming antigen in vivo ATCC Info http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=TIB-39&Template=cellBiology In this paper they use B16 (melanoma cells from C57BL/6J mice) pulsed with the SIINFEKL peptide along with OT-1 CD8+ T cells from C57BL/6 mice. http://jem.rupress.org/content/207/1/223.full.pdf B16 melanoma cells have the H2-Kb haplotype Both C57BL/6 and C57BL/6J have the H2-Kb haplotype I wasn't sure exactly what OT-1 CD8+ T cells are. In the paper they state "OT-1 CD8+ T cells, harvested from spleens of C57BL/6 mice, expressa transgene encoding a TCR that specifically recognizes SIINFEKL peptide bound to MHC-I H-2kb (Hogquist et al., 1994)." From what I understand, they are doing something a little different with their "OT-1 CD8+" cells that we won't need to do. We will just need to immunize our C57BL/6 mice like we stated before. They don't say whether they use B16-F0, B16-F1, or B16-F10 cells. In our lab we have B16-F1 and B16-F10 cells. Do you have a preference about which we should use? Does it matter? This paper mentions something about H-2Kd Ova specificity CD8+ T cell concentration determines their efficiency in killing cognate antigen–expressing syngeneic mammalian cells in vitro and in mouse tissues (2010 paper from Columbia University) http://www.wikigenes.org/e/ref/e/1534037.html
 * CD8+ T cell concentration determines their efficiency in killing cognate antigen–expressing syngeneic mammalian cells in vitro and in mouse tissues** (2010 paper from Columbia University)

These other papers also characterize T cell and B16 cell interactions Moore et al., 1988; Ochalek et al., 1988; Snyder et al., 2003; Regoes et al., 2007) and in vivo (Dobrzanski et al., 2001; Regoes et al., 2007).