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Results: 5

1.
Fig. 1.

Fig. 1. From: Peptide vaccines prevent tumor growth by activating T cells that respond to native tumor antigens.

Peptide variants elicit a range of antitumor immunity. Mice were vaccinated with BV-infected insect cells expressing peptide-MHC molecules 7 and 14 d before challenge with CT26 tumor cells. Mice were killed and scored when the tumor grew to 10 mm. Survival curves of mice vaccinated with the indicated peptides were compared with βgal-vaccinated mice using a log-rank test: ***, P < 0.0001; **, P = 0.0021. The survival curve of the AH1-vaccinated mice (gray dashed line) closely overlaps with the survival curve of the βgal vaccinated (gray solid line). The bold residues indicate where the variant peptides differ from the residues in the AH1 peptide.

Kimberly R. Jordan, et al. Proc Natl Acad Sci U S A. 2010 March 9;107(10):4652-4657.
2.
Fig. 5.

Fig. 5. From: Peptide vaccines prevent tumor growth by activating T cells that respond to native tumor antigens.

Vaccination with protective peptide variants elicits more CTLs that kill AH1-loaded target cells. (A) Splenocytes from Fig. 3 were stimulated with AH1 peptide in the presence of CD107a (LAMP-1) antibody and stained with CD8 and dump antibodies. The number of CD107a+ CD8+ cells was determined by flow cytometry. (B) Sorted AH1-tet+ CD8+ cells (as in Fig. 4D) were incubated with CFSE-labeled peptide-loaded target cells. Three days later, the number of live (7AAD) target cells was determined by flow cytometry. The number of live βgal peptide-loaded (CFSElow) or AH1 peptide-loaded (CFSEhi) target cells are indicated. (C) AH1-tet+ CD8+ cells were incubated at different effector to target ratios (E:T) as in B. Groups were compared using a Student’s t-test (n = 3; **, P = 0.006; *, P = 0.02).

Kimberly R. Jordan, et al. Proc Natl Acad Sci U S A. 2010 March 9;107(10):4652-4657.
3.
Fig. 3.

Fig. 3. From: Peptide vaccines prevent tumor growth by activating T cells that respond to native tumor antigens.

Vaccination with protective peptide variants elicits more endogenous AH1-specific T cells. (A) Splenocytes from mice vaccinated with the indicated peptides were stained with AH1-tet, CD8, and dump antibodies. The number (left axis, black bars) and percentage (right axis, white bars) of AH1tet+CD8+ cells was determined and compared with the number of cells in mice vaccinated with the AH1 peptide using a Student’s t test (n = 3; P ≤ 0.0151 for all peptides). Error bars represent SEM. (B) Splenocytes from A were stained with the corresponding peptide variant-tetramer (variant-tet), CD8, and dump antibodies and analyzed as in A. (C) Splenocytes from A were costained with both AH1-tet (y axis) and the corresponding variant-tet (x axis), CD8, and dump antibodies. The percentage of dumpCD8+ dual tet+ (cross-reactive, Upper Right) or variant-tet+ (Lower Right) cells is indicated. (D) The percentage of variant-tet+ cells that bind to AH1-tet from C.

Kimberly R. Jordan, et al. Proc Natl Acad Sci U S A. 2010 March 9;107(10):4652-4657.
4.
Fig. 4.

Fig. 4. From: Peptide vaccines prevent tumor growth by activating T cells that respond to native tumor antigens.

Vaccination with protective peptide variants elicits more CTLs that produce IFNg in response to the AH1 peptide. (A) Splenocytes from Fig. 3 were stimulated with AH1 peptide and stained with IFNg, CD8, and dump antibodies. The number of CD8+IFNg+ cells was determined by flow cytometry. (B) The number of CD8+IFNg+ cells from A was divided by the number of CD8+AH1tet+ cells from Fig. 3A to determine the percentage of AH1-specific T cells that produced IFNg. (C) Splenocytes from Fig. 3 were stimulated with the corresponding peptide variant and analyzed as in A. (D) 2.5 × 104 AH1tet+ cells isolated from mice vaccinated with the F1A5 or WMF peptides were incubated with increasing concentrations of the AH1 peptide or the corresponding peptide variant. The concentration of IFNg secreted was determined by ELISA 24 h later (performed in duplicate, n = 3; *, P = 0.03; **, P = 0.01).

Kimberly R. Jordan, et al. Proc Natl Acad Sci U S A. 2010 March 9;107(10):4652-4657.
5.
Fig. 2.

Fig. 2. From: Peptide vaccines prevent tumor growth by activating T cells that respond to native tumor antigens.

Peptide variants bind to Ld similarly and stimulate T cells expressing the CT-TCR more effectively than the native AH1 peptide. (A) The amount of competing peptide required to reduce the Ld binding by 50%, the EC50 value, was determined by fitting the relative RU values to a sigmoidal dose-response curve using a one site-competition equation (Prism Graphpad). (B) Insect cells were infected with recombinant baculovirus encoding membrane-bound peptide-MHC complexes and stained with an Ld antibody and increasing concentrations of soluble fluorescent multimerized CT-TCR. To control for infection differences, the CT-TCR mean fluorescence intensity (MFI) was determined at a constant level of MHC-expression. The CT-TCR MFIs at 500 μg/mL are listed. (C) CFSE-labeled splenocytes from CT-TCR Tg mice were incubated with increasing concentrations of peptide in vitro. Proliferation was determined by CFSE dilution of CD8+ T cells. (D) CFSE-labeled Thy1.2+ splenocytes from CT-TCR Tg mice were transferred into naïve Thy1.1+ BALB/c mice and vaccinated 1 day later with infected insect cells expressing the indicated peptide-MHC molecules. Proliferation was determined by CFSE dilution of Thy1.2+ CD8+ T cells.

Kimberly R. Jordan, et al. Proc Natl Acad Sci U S A. 2010 March 9;107(10):4652-4657.

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