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. 2010 Oct;88(4):779–789. doi: 10.1189/jlb.0410237

Figure 6. Peptide treatment results in death of TLR-activated B cells and requires TCR specificity.

Figure 6.

Purified B, CD4+ T, and CD8+ T cells were isolated from C57BL/6 murine spleens using magnetic bead separation (Miltenyi Biotec). Isolated cells, combined as indicated, were cultured (106 cells/ml) in the presence of CpG-ODN, with or without peptides (Table 2), including TPP, HB, LB, or OVA 323–339 at an E:T ratio of 1:5. The cells were harvested and stained with 15G4-FITC, anti-mouse CLIP/I-Ab, and PE-conjugated anti-mouse B220, antibody to the B cell molecule B220. (A, left) The number of live B220+, CLIP+ population (*P<0.001). (A, right) The percent of dead cells resulting from treatments as indicated (*P<0.001). (B) B cells were isolated from C57BL/6 spleen and cultured with or without purified CD4+ T cells isolated from OT-2 transgenic mice expressing only TCRs specific for the OVA peptide 323–339. Cells were cultured with and/or without CD8+ T cells derived from C57Bl6. All cells were cultured at 106 cells/ml in the presence of CpG, with or without TPP or OVA 323–339 at an E:T ratio of 1:5 (*P <0.001). Act. B, Activated B cell.