Anti tigit mab

The BMSCs and NK cells from patients with MDS were co-cultured at a ratio of 4:1 and grouped according to the addition of different immune checkpoint inhibitors, including anti-TIGIT mAb (R&D Systems) and the CD226 agonist (WO 2020/023312 A1, Additional file 1: Fig. S1). After 6 days of incubation, FCM was used to detect the changes in NK cell function before and after co-cultivation. At the same time, the MDS cell line (SKM-1) was also co-cultured with NK cells at the effector cell: target cell ratios(E:T ratio) of 1:1, 5:1, and 10:1 for 6 h after co-cultivation using the Annexin V Cell Apoptosis Analysis Kit (BD Biosciences).

The mouse cervical cancer cell line U14 was obtained from the Academy of Medical Sciences (Beijing, China). U14 cells were cultured in DMEM nmented with 10% foetal bovine serum (all from Gibco, Grand Island, NY, USA), 50 U/mL penicillin and 50 mg/mL streptomycin (all from Solarbio Science & Technology, Beijing, China). CD8⁺ T cells were purified from PBMCs through positive selection using a kit (Milteny Biotec, Bergisch Gladbach, Germany). CD8⁺ T cells were stimulated with an anti-CD3/CD28 antibody (Stemcell, Canada) in T cell expansion medium (Stemcell, Canada). Activated CD8⁺ T cells were treated with 5 μg/mL CD155-Fc (R&D Systems), or 10 μg/mL CD155-Fc. Activated CD8⁺ T cells were cocultured with tumour cells at a 10:1 ratio. Next, 10 μg/ml anti-PD-1 mAb or 5 μg/ml anti-TIGIT mAb (R&D Systems) were added to the cells. We used α-human IgG1 (R&D Systems) as an isotype control. After 48 h, CD8⁺ T cells were collected to determine cytokine production using the T cell function assay.