Figure 1

Cytotoxicity of WT1-specific and HLA-A*24:02-restricted CTLs against podocytes. (A) Expression of HLA-A24:02 on the HLA-A*24:02 gene-transduced mouse podocyte cell line, MPC-5 (MPC-5-A24). Flow cytometric analysis was performed using anti-HLA-A24:02 monoclonal antibody (One Lambda, Canoga Park, CA, USA). (B) Expression of HLA class I in the glomerulus of a HLA-A*24:02-transgenic mouse (original magnification ×400). Immunohistochemistry was performed using an anti-HLA class I framework monoclonal antibody (Abcam, Cambridge, UK). (C) Expression of WT1 in the HLA-A*24:02 gene-transduced mouse podocyte cell line, MPC-5-A24. MPC-5-A24 cells and LCL were stained with a rabbit anti-human and mouse WT1 polyclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA) (original magnification ×400). Notably, WT1 is abundantly expressed in MPC-5-A24 cells but not LCL. (D) Expression of WT1 in the glomerulus of a HLA-A*24:02-transgenic mouse (original magnification ×400). Immunohistochemistry was performed using a rabbit anti-human and mouse WT1 polyclonal antibody (Santa Cruz Biotechnology). (E) Cytotoxicity of WT1-specific and HLA-A*24:02-restricted CTLs against MPC-5, MPC-5-A24, and HLA-A*24:02-positive LCLs in the presence or absence of WT1 peptide at various effector:target cell ratios.