T-cell clone 4G11 efficiently recognizes B-cell malignancies while sparing non–B-cell lineages. (A-B) T-cell clones (A) 4G11 and (B) 3C10 were coincubated with primary samples of malignant B cells, including CLL, ALL, MCL, and MM. HLA allotype of each sample is indicated. Controls included BOB1⁻ K562-B7 cells and BOB1-expressing LCL-JY. Shown is 1 representative experiment of 2 independent experiments. (C-D) T-cell clones (C) 4G11 and (D) 3C10 were incubated with K562-B7 and K562-A2 cells alone or transduced to express BOB1 (K562-B7 + BOB1 or K562-A2 + BOB1, respectively) or K562-B7 and K562-A2 cells pulsed with 1 µM BOB1₄₄ or BOB1₂₄₅ peptide, respectively. (E-J) T-cell clone 4G11 was cocultured with different HLA-B7⁺ healthy primary or activated cell subsets. (E) Fibroblasts (FBs) from 3 different individuals were either left untreated or cultured for 4 days in 200 IU/ml IFN-γ (FB + IFN-γ) before coculture. Primary bronchial epithelial cells were cultured under (F) non–air-exposed or (G) air-exposed condition before being used as stimulator cells. Cells had also been cultured in the absence (−IFN-γ) or presence (+IFN-γ) of 100 IU/ml IFN-γ for 2 days before coculture. (H) Clone 4G11 was coincubated with various HLA-B7⁺ BOB1⁻ nonhematopoietic tumor cell lines. (I) Primary CD19⁺, CD4⁺, CD14⁺, and CD34⁺ cells were isolated from PBMCs of healthy individuals (UDR, UHK, AFD, and ALY) and cocultured in different responder-to-stimulator (R:S) ratios. Activated B cells (CD19 [CD40L]) were generated by stimulating CD19⁺ B cells with CD40L. Activated T cells were generated from phytohemagglutinin-stimulated PBMCs. (J) Immature (imDC) and mature (mDC) dendritic cells were monocyte derived. (K) T-cell clone 4G11 was cocultured with a panel of BOB1-expressing B-LCLs expressing a wide variety of HLA class I and II molecules. HLA-B7 status of each B-LCL is indicated with + (positive) or – (negative). IFN-γ production was measured after 18 hours of coculture. Shown are means with standard deviations of 1 experiment carried out in duplicate. Representative data from ≥2 independent experiments.