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. Author manuscript; available in PMC: 2013 Oct 28.
Published in final edited form as: J Immunol. 2011 Jan 12;186(4):10.4049/jimmunol.1003225. doi: 10.4049/jimmunol.1003225

FIGURE 5.

FIGURE 5

Open-CD3 outlasts TCR engagement by APCs. A, OT-I Rag−/− β2m−/− preselection DP thymocytes were cocultured for 30 min at 37°C with T2-Kb APCs that had been preloaded with either pFARL (null peptide), pOVA (antigenic peptide), or no peptide plus soluble anti-CD3ε mAb 2C11. Then, cocultures were lysed and subjected to the CD3-PD assay. CD3-PD samples were visualized for their content of 2C11, MHC-I H chain, and CD3ζ by WB. Open-CD3 inductions achieved by antigenic and 2C11 stimulation were calculated relative to the amounts of open-CD3 found in basal conditions (pFARL). B, T1 hybridoma cells were cocultured for 30 min with P-815 APCs that had been preloaded with either no peptide (Ø), pSYIP (antigenic peptide), or no peptide plus soluble anti-CD3ε mAb 2C11. Then, cocultures were lysed and subjected to the CD3-PD assay or H2-Kd immunoprecipitation with the mAb SF1-1.1. CD3-PD samples were visualized for their content of 2C11, MHC-I H chain, and CD3ζ by WB. Open-CD3 inductions achieved by antigenic and 2C11 stimulation were calculated relative to the amounts of open-CD3 found in basal conditions (Ø).