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. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: J Immunol. 2013 Jul 5;191(3):10.4049/jimmunol.1300658. doi: 10.4049/jimmunol.1300658

Figure 2.

Figure 2

20.1 mAb specifically stimulates diverse Vγ2Vδ2 T cells in a Vγ2Vδ2 TCR-dependent manner. (A) 20.1 mAb specifically stimulates the expansion of Vγ2Vδ2 T cells from blood mononuclear cells. PBMC from five donors were cultured with media, HMBPP (100 nM), a control mAb (10 μg/ml), or the 20.1 mAb (10 μg/ml). On day 3, IL-2 was added to 1 nM. After 9 d, cells were stained with the indicated mAbs and analyzed by flow cytometry. (B) Specific stimulation of Vγ2Vδ2 T cell clones by the 20.1 mAb. CD8αα+/CD4- CD8- (solid bars) and CD4+ (open bars) T cell clones expressing different V genes were stimulated with the 20.1 mAb (1 μg/ml) or PHA-P (1:1000) with Va2 APC and [3H]-thymidine incorporation measured at day 2. Stimulation index was calculated as the ratio between the 20.1 and control mAb response. PHA stimulation indices were >2.5 for all clones and averaged 12.6-fold. (C) 20.1 mAb stimulation is mediated by the Vγ2Vδ2 TCR. The DBS43 Vγ2Vδ2 TCR transfectant and the parent β− J.RT3-T3.5 cell line line were incubated with mitomycin C-treated Va2 APC and either the 20.1 mAb or HMBPP for 24 h. The supernatants were then harvested and used to stimulate the IL-2-dependent proliferation of HT-2 T cells.