Anti cd28 mab clone cd28

The cell staining procedure used in this study was described previously (Wu et al., 2018a (link)). For intracellular cytokine staining, the cells were restimulated for 12 h with 10 g/ml CagA (Linc-Bio Science, Shanghai, China), 10 μg/ml VacA (Linc-Bio Science, Shanghai, China), anti-CD3 mAb (Clone UCHT1, BD), anti-CD28 mAb (Clone CD28.2, BD) and 3 μg/ml brefeldin A (eBioscience, CA, USA). Intracellular cytokines were stained using the intracellular fixation/permeabilization buffer set (eBioscience, CA, USA). Flow cytometric analysis was performed on FACS Canto II (BD, NJ, USA), and the data were analyzed using FlowJo software (Tree Star). The following anti-human antibodies were purchased from eBioscience, BD Biosciences or Biolegend (CA, USA): CD3 (Clone UCHT1, BD), CD4 (Clone L200, BD), CD8 (Clone RPA-T8, BD), CD45RO (Clone UCHL1, BD), CD69 (Clone FN50, Biolegend), CD103 (Clone Ber-ACT8, Biolegend), CCR7 (Clone 150503, BD), CCR9 (Clone L053E8, Biolegend), CXCR3 (Clone 1C6/CXCR, BD), E-Cadherin (Clone DECMA-1, Biolegend), TNF (Clone MAb11, eBioscience), IFN-γ (Clone 4S.B3, eBioscience), IL-17a (Clone BL168, Biolegend), T-bet (Clone O4-46, BD), and Blimp1 (Clone 6D3, BD), ZAP70 Phospho (Tyr319) (Clone 1503310, Biolegend).

The procedure for cell staining in this study was described previously (53 (link)). For IL-9 staining, MAIT cells were restimulated for 12 h with 1 μg/ml anti-CD3 mAb (Clone UCHT1, BD), 1 μg/ml anti-CD28 mAb (Clone CD28.2, BD) and 3 μg/ml brefeldin A (eBioscience, CA, USA). IL-9 staining was performed with the intracellular fixation/permeabilization buffer set (eBioscience, CA, USA). Flow cytometric analysis was performed on FACS Canto II (BD, NJ, USA), and data were analyzed using FlowJo software (Tree Star). Involved anti-human antibodies were purchased from eBioscience, BD Biosciences or Biolegend (CA, USA): CD3 (Clone UCHT1, BD), CD161 (Clone HP-3G10, Biolegend), OX40 (Clone Ber-ACT35, Biolegend), OX40L (Clone ik-1, BD), CD11c (Clone B-ly6, BD), TCR Vα7.2 (Clone 3C10, Biolegend), and IL-9 (Clone Ber-ACT8, Biolegend).