Induction of Immune Thrombocytopenia in Mice

After 1-week acclimation, mice were randomly assigned into four groups (n = 6): Control group, ITP group, ITP + vector group and ITP + sh-HLA-DRB5 group. Before establishment of ITP murine models, guinea pig anti-mouse platelet serum (GP-APS) that can be used to induce ITP was customized and purchased from Cloud-Clone Corp. (Wuhan, China). Initially, mice received injection of serum containing GP-APS, and then PLT was removed from the circulation by the fixed phagocyte system [18 (link)]. In this study, GP-APS was diluted with saline (G4702, Servicebio, Wuhan, China) in a ratio of 1:4 to prevent erythrocyte adsorption [19 (link)]. Then, besides mice from Control group that were given the same volume of saline, mice from other groups were intraperitoneally injected with 100 μL GP-APS every other day. Meanwhile, mice from ITP + sh-HLA-DRB5 group or ITP + vector group were injected with 50 μL diluted sh-HLA-DRB5 adenovirus or negative control by tail vein once a week for 2 weeks. After 15 days, all mice were euthanized by cervical disarticulation under anesthesia (50 mg/kg pentobarbital sodium, P-010; Sigma-Aldrich, St Louis, MO, USA).