Modulating T Cell Activation Pathways

CD4⁺ T cells isolated from spleen and LN using CD4 microbeads
(Miltenyi). 1×10⁶ T cells cultured with platebound
α-CD3 (2C11) and α-CD28 (37.51) from BioXcell at 1μg/mL
each for 48 or 66 hrs. Inhibitors purchased and used as follows: ZSTK474
(1μM, Sigma), Rapamycin (10nM, Selleckchem), NFAT inhibitor (10μM,
Tocoris), Mek inhibitor PD0325901 (100nM, Peprotech) and Erk inhibitor FR180204
(10μM, Tocoris). In vitro T_R suppression
assays were performed as previous described (20 (link)). Chemotaxis assay performed as previously described (21 (link)).

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Sullivan J.M., Höllbacher B, & Campbell D.J. (2018). Dynamic expression of Id3 defines the stepwise differentiation of tissue-resident regulatory T cells. Journal of immunology (Baltimore, Md. : 1950), 202(1), 31-36.

Publication 2018

CD4 microbeads α-CD28 (37.51) ZSTK474 Rapamycin

Assay Cd4 t cells Chemotaxis Fr180204 Inhibitors Mek inhibitor pd0325901 Microbeads Rapamycin Spleen T cells Zstk474

Corresponding Organization : University of Washington

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Variable analysis

independent variables

α-CD3 (2C11) and α-CD28 (37.51) from BioXcell at 1μg/mL
ZSTK474 (1μM, Sigma)
Rapamycin (10nM, Selleckchem)
NFAT inhibitor (10μM, Tocoris)
Mek inhibitor PD0325901 (100nM, Peprotech)
Erk inhibitor FR180204 (10μM, Tocoris)

dependent variables

T cell activation and proliferation (measured through in vitro T_R suppression assays)
T cell chemotaxis (measured through chemotaxis assays)

control variables

CD4+ T cells isolated from spleen and LN using CD4 microbeads (Miltenyi)
1×10^6 T cells cultured for 48 or 66 hrs

positive controls

None specified

negative controls

None specified

Annotations

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