Figure 4.

Blockade of the BTLA/HVEM signaling does not affect the degranulation of in vitro expanded γδ T cells. (A) Representative histograms showing CD107a expression in the γδ T cells. Gray lines indicate the isotype control. (B) Cells were expanded in the indicated culture condition for 14 days. In the presence of anti‐human CD107a mAb, in vitro expanded γδ T cells with the indicated inactivated cells (on top) were incubated overnight with WT Jurkat (upper panel) or HVEM^low Jurkat (lower panel) cells. Representative data are shown out of three to four independent experiments using distinct donors. (C) The percentages of CD107a⁺ γδ T cells in the anti‐PD‐L1 treated group were normalized by dividing by that of the untreated group. Data were shown as mean ± SEM of three to four independent experiments using distinct donors. Paired t test was used (*p < .05). Each P value is .0190, .0345, and .0103 from left to right. (D) The gating strategy to identify intracellular IFN‐γ is depicted. (E) Expanded γδ T cells with the indicated inactivated cells (on top) were incubated overnight with WT Jurkat cells in the presence of BTLA/HVEM blocking peptides and anti‐PD‐L1 Ab or adequate controls for IFN‐γ intracellular staining. (F) The percentages of IFN‐γ⁺ γδ T cells were summarized. N = 2. BTLA, B‐ and T‐lymphocyte attenuator; HVEM, herpes virus entry mediator; IFN‐γ, interferon; mAB, monoclonal antibody; PD‐L1, programmed death‐ligand 1; WT, wild‐type