Adoptive Transfer of Pmel-1 and OT-II T Cells

Pmel-1 (B6.Cg-/Cy Tg [TcraTcrb] 8Rest/J), Rag2^−/−, OT-II (B6.Cg-Tg (TcraTcrb)425Cbn/J), and C57BL/6 mice were obtained from the Jackson Laboratory. C57BL/6 male mice of 6-8 weeks of age were used as recipient hosts for adoptive transfer unless otherwise indicated. We crossed Pmel-1 with Ly5.1 mice (B6.SJL-Ptprc^aPepc^b/BoyJ) to obtain Pmel-1 Ly5.1 mice. We crossed OT-II with Rag2^-/- to obtain OT-II Rag^-/- mice. All mice were maintained under specific pathogen–free conditions. B16 (H-2D^b), a murine melanoma, transduced as previously described31 (link) to express gp100 with human residues at position 25-27; EGS -> KVP. Mel624 was obtained from ATCC and Platinum-E ecotropic packaging cells were obtained from Cell Biolabs. Cell lines and maintained in DMEM media with 10% FBS, 1% glutamine and 1% penicillin-streptomycin.

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Eil R., Vodnala S.K., Clever D., Klebanoff C.A., Sukumar M., Pan J.H., Palmer D.C., Gros A., Yamamoto T.N., Patel S.J., Guittard G.C., Yu Z., Carbonaro V., Okkenhaug K., Schrump D.S., Linehan W.M., Roychoudhuri R, & Restifo N.P. (2016). Ionic immune suppression within the tumour microenvironment limits T cell effector function. Nature, 537(7621), 539-543.

Publication 2016

Pmel-1 OT-II C57BL/6 Mel624 Platinum-E ecotropic packaging cells

Adoptive transfer C57bl mice Cell Cell lines Glutamine Human Male Melanoma Murine Penicillin Platinum Pmel Rag2 Specific pathogen free Streptomycin

Corresponding Organization : National Cancer Institute

Other organizations : Babraham Institute

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Variable analysis

independent variables

Pmel-1 (B6.Cg-/Cy Tg [TcraTcrb] 8Rest/J)
Rag2-/- mice
OT-II (B6.Cg-Tg (TcraTcrb)425Cbn/J) mice
C57BL/6 male mice of 6-8 weeks of age

dependent variables

Adoptive transfer outcomes

control variables

Mice were maintained under specific pathogen–free conditions
Cell lines were maintained in DMEM media with 10% FBS, 1% glutamine and 1% penicillin-streptomycin

positive controls

C57BL/6 male mice of 6-8 weeks of age used as recipient hosts for adoptive transfer

negative controls

Not explicitly mentioned

Annotations

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