Research 3-20-12

2015-01-13

azim58 - 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/Defa
ult.aspx?ATCCNum=TIB-39&amp;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.
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://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

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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).

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