Fig. 5.
Fig. 5. Vaccine-induced expansion of tumor-specific T cells is enhanced by NST and administration of DLI. / BALB/c mice (A-D) or B10.D2→BALB/c chimeras generated on day 0 by NST (E-H, see legend to Figure 1) received no treatment (A-B), autologous tumor cell vaccine (C-F), or 20 million B10.D2 spleen cells followed by autologous tumor cell vaccine (G-H). Six weeks after vaccine administration, recipients' spleens were harvested, and CD8+ T cells were purified and incubated for 7 days with AH1 peptide in complete medium supplemented with 10 U/mL IL-2. AH1-specific T cells in culture were identified by 2-color flow cytometry after staining with CyChrome-conjugated antibody to mouse CD8 and Ld-Ig dimers loaded with AH1 peptide, followed by biotinylated goat antibody to mouse IgG1 and phycoerythrin-conjugated avidin (A,C,E,G). As a negative control, T cells were also stained with Ld-Ig dimers loaded with an irrelevant peptide from mouse β-galactosidase (Ld-Ig/β-gal; B,D,F,H).

Vaccine-induced expansion of tumor-specific T cells is enhanced by NST and administration of DLI.

BALB/c mice (A-D) or B10.D2→BALB/c chimeras generated on day 0 by NST (E-H, see legend to Figure 1) received no treatment (A-B), autologous tumor cell vaccine (C-F), or 20 million B10.D2 spleen cells followed by autologous tumor cell vaccine (G-H). Six weeks after vaccine administration, recipients' spleens were harvested, and CD8+ T cells were purified and incubated for 7 days with AH1 peptide in complete medium supplemented with 10 U/mL IL-2. AH1-specific T cells in culture were identified by 2-color flow cytometry after staining with CyChrome-conjugated antibody to mouse CD8 and Ld-Ig dimers loaded with AH1 peptide, followed by biotinylated goat antibody to mouse IgG1 and phycoerythrin-conjugated avidin (A,C,E,G). As a negative control, T cells were also stained with Ld-Ig dimers loaded with an irrelevant peptide from mouse β-galactosidase (Ld-Ig/β-gal; B,D,F,H).

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