Journal: Cancer Immunology Research

Article Title: IDO Vaccine Ablates Immune-Suppressive Myeloid Populations and Enhances Antitumor Effects Independent of Tumor Cell IDO Status

doi: 10.1158/2326-6066.CIR-21-0457

Figure Lengend Snippet: IDO vaccine enhances immune-mediated antitumor effects of tumor antigen–specific vaccination and prolongs survival in the B16F10 tumor model. Gating strategy and IDO pentamer specificity in Supplementary Fig. S2. A, Schematic of the treatment schedule in the tumor model. On day 10 of tumor growth, B16F10 tumor-bearing mice were given IDO vaccine (IDO Vax) with the gp100 25–33 peptide vaccine, along with PADRE (20 μg/mouse) and QuilA (10 μg/mouse) subcutaneously, every 7 days for a total of three doses. Tumor growth and survival were measured. B, Average tumor volume in mice following treatment (* vs. untreated; green * vs. IDO Vax; red * vs. gp100). C, Percent survival of mice depicted by the Kaplan–Meier plot. D, SK plot showing tumor volume and survival for each mouse at different days. Data are shown as an average of two independent experiments ( n = 10–18 per group). Error bars indicate the SEM. For tumor growth, statistical analysis was performed by unpaired, one-tailed Student t test. Survival in various groups was compared using log-rank (Mantel–Cox) tests. *, P ≤ 0.05; and **, P ≤ 0.01. E–K, C57BL/6J mice ( n = 5–8 per group) were treated as in A , except 3 days after second vaccination, mice were sacrificed, and tumors were harvested for immune response study. The frequency of B16F10 tumor-infiltrating cells was determined. Total ( E ), IFNγ + and GB + ( F ), CD40L + ( G ), Penta-IDO + ( H ), IFNγ + penta-IDO + ( I ), Dextra-gp100 + ( J ), IFNγ + dextra-gp100 + ( K ) CD8 + T cells were measured by flow cytometry. Data are shown from one representative experiment of two independent experiments. Error bars indicate the SEM. Statistical analysis was performed by unpaired, one-tailed Student t test. NS: nonsignificant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.

Article Snippet: The CTL epitope from human gp100 25–33 (KVPRNQDWL; an altered peptide ligand vaccine for mice (100 μg/mouse; Celtek Bioscience) for the B16F10 tumor model or HPV16 E7 49–57 (RAHYNIVTF) for TC-1 tumor model [9-amino acid (aa) peptide, 100 μg/mouse] mixed with synthetic T-helper epitope PADRE (13 aa peptide, aK-Cha-VAAWTLKAAa, where “a” is D-alanine and Cha is L-cyclohexylalanine, 20 μg/mouse; both from Celtek Bioscience) and QuilA adjuvant (10 μg/mouse; Brenntag) were used as the model vaccine (subcutaneous injections, as described below), which contains both CD8 and CD4 epitopes in all studies ( ).

Techniques: One-tailed Test, Flow Cytometry

Journal: Cancer Immunology Research

Article Title: IDO Vaccine Ablates Immune-Suppressive Myeloid Populations and Enhances Antitumor Effects Independent of Tumor Cell IDO Status

doi: 10.1158/2326-6066.CIR-21-0457

Figure Lengend Snippet: IDO vaccine reduces frequency of Tregs and enhances therapeutic ratio in the TME. C57BL/6J mice were treated as in Fig. 1A and , except 3 days after second vaccination, mice were sacrificed, and tumors were harvested for immune response study. The frequency of B16F10 tumor-infiltrating CD4 + ( A ), FoxP3 + CD4 + ( B ), CD8 + /Tregs ( C ), Dextra-gp100 + CD8 + /Tregs ( D ), and Penta-IDO + CD8 + /Tregs ( E ) was determined by flow cytometry. The frequency of TC-1 tumor-infiltrating CD4 + ( F ), FoxP3 + CD4 + ( G ), CD8 + /Tregs ( H ), Dextra-E7 + CD8 + /Tregs ( I ), and Penta-IDO + CD8 + /Tregs ( J ) were also measured by flow cytometry. Data are shown as an average of two independent experiments ( n = 8–13 per group). Error bars indicate the SEM. Statistical analysis was performed by unpaired, one-tailed Student t test. NS, nonsignificant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.

Article Snippet: The CTL epitope from human gp100 25–33 (KVPRNQDWL; an altered peptide ligand vaccine for mice (100 μg/mouse; Celtek Bioscience) for the B16F10 tumor model or HPV16 E7 49–57 (RAHYNIVTF) for TC-1 tumor model [9-amino acid (aa) peptide, 100 μg/mouse] mixed with synthetic T-helper epitope PADRE (13 aa peptide, aK-Cha-VAAWTLKAAa, where “a” is D-alanine and Cha is L-cyclohexylalanine, 20 μg/mouse; both from Celtek Bioscience) and QuilA adjuvant (10 μg/mouse; Brenntag) were used as the model vaccine (subcutaneous injections, as described below), which contains both CD8 and CD4 epitopes in all studies ( ).

Techniques: Flow Cytometry, One-tailed Test

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Peptide vaccine formulation controls the duration of antigen presentation and magnitude of tumor-specific CD8+ T cell response

doi: 10.4049/jimmunol.1700467

Figure Lengend Snippet: L-Tyrosine is a potent vaccine adjuvant for the induction of CD8+ T cell responses. A) Pmel-1 T cell level as a percentage of CD8 + T cells in the blood of mice at different time point. B) IFN-γ and C) granzyme B production by pmel-1 T cells. D) Memory phenotype of pmel-1 T cells on day 300 post vaccination. Mice received 8×105 pmel-1 T cells and indicated treatments on day 0. Peptide dose was 50 μg per mouse. Data are shown as the mean ± s.e.m. Statistical differences between the two groups were determined by student t-test. n = 5 mice per group per experiment. Data are representative of 3 independent experiments.*: P< 0.05, **: P< 0.01, ns: not significant.

Article Snippet: The synthetic, high-affinity H-2D b -restricted heteroclitic mouse gp100 25–33 peptide (KVPRNQDWL) and H-2K b –restricted chicken OVA-I 257–264 peptide (SIINFEKL) were purchased from Peptides International (Louisville, KY) at a purity >95%.

Techniques: Adjuvant

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Peptide vaccine formulation controls the duration of antigen presentation and magnitude of tumor-specific CD8+ T cell response

doi: 10.4049/jimmunol.1700467

Figure Lengend Snippet: Neutrophils do not contribute to T cell priming after L-Tyrosine vaccine. Anti-Ly6G Ab was given 2 days prior to vaccination and every 3 days afterward. A) Pmel-1 T cells level in the blood after indicated treatments were shown. The depletion of Ly6G population was confirmed in peripheral blood (B) and the skin at vaccine sites (C) on day 4 after vaccination. Mice received 8×105 pmel-1 T cells and indicated treatments on day 0.n = 5 mice per group. Data are representative of 2 independent experiments.

Article Snippet: The synthetic, high-affinity H-2D b -restricted heteroclitic mouse gp100 25–33 peptide (KVPRNQDWL) and H-2K b –restricted chicken OVA-I 257–264 peptide (SIINFEKL) were purchased from Peptides International (Louisville, KY) at a purity >95%.

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Peptide vaccine formulation controls the duration of antigen presentation and magnitude of tumor-specific CD8+ T cell response

doi: 10.4049/jimmunol.1700467

Figure Lengend Snippet: L-Tyrosine adjuvant activity does not require inflammasome activation. A) No difference in T cell responses between wild-type and ASC KO groups was found, regardless of vaccine formulations. B) L-Tyrosine adjuvant activity requires peptide co-precipitation. Mice received 8×105 pmel-1 T cells and indicated treatments on day 0. n= 5 mice per group. Data are representative of 3 independent experiments. WT: wild type, ASC KO: ASC knockout.

Article Snippet: The synthetic, high-affinity H-2D b -restricted heteroclitic mouse gp100 25–33 peptide (KVPRNQDWL) and H-2K b –restricted chicken OVA-I 257–264 peptide (SIINFEKL) were purchased from Peptides International (Louisville, KY) at a purity >95%.

Techniques: Adjuvant, Activity Assay, Activation Assay, Knock-Out

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Peptide vaccine formulation controls the duration of antigen presentation and magnitude of tumor-specific CD8+ T cell response

doi: 10.4049/jimmunol.1700467

Figure Lengend Snippet: L-Tyrosine formulation extends the duration of antigen presentation. A) A schematic of the experiment design. All mice were treated with indicated treatments on day 0. At indicated time points, 2×106 CFSE labeled pmel-1 CD8+ T cells were transferred to hosts. 72 hours post transfer, vaccine draining lymph nodes were harvested and CFSE dilution of pmel-1 T cells was measured by flow cytometry. n = 3 - 5 mice per group. Data are representative of 2 independent experiments.

Article Snippet: The synthetic, high-affinity H-2D b -restricted heteroclitic mouse gp100 25–33 peptide (KVPRNQDWL) and H-2K b –restricted chicken OVA-I 257–264 peptide (SIINFEKL) were purchased from Peptides International (Louisville, KY) at a purity >95%.

Techniques: Formulation, Immunopeptidomics, Labeling, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Peptide vaccine formulation controls the duration of antigen presentation and magnitude of tumor-specific CD8+ T cell response

doi: 10.4049/jimmunol.1700467

Figure Lengend Snippet: Anti-tumor efficacy of different vaccine formulations regimens. Mice received 8×105 pmel-1 T cells and indicated treatments on day 0. A) Pmel-1 T cell response after different vaccine formulations were followed and B) their function at day 7 post vaccination was shown (mean ± s.e.m.). C) Anti-tumor efficacy of corresponding groups of the same experiment. n = 10 – 20 mice/group. Peptide dose was 50ug per mice. Data pooled from 2 independent experiments. Differences in survival among groups were compared using log-rank test. *: P< 0.05, **: P< 0.01, ns: not significant, n/a: not available.

Article Snippet: The synthetic, high-affinity H-2D b -restricted heteroclitic mouse gp100 25–33 peptide (KVPRNQDWL) and H-2K b –restricted chicken OVA-I 257–264 peptide (SIINFEKL) were purchased from Peptides International (Louisville, KY) at a purity >95%.

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Peptide vaccine formulation controls the duration of antigen presentation and magnitude of tumor-specific CD8+ T cell response

doi: 10.4049/jimmunol.1700467

Figure Lengend Snippet: T cell responses from the endogenous repertoire. A) Endogenous gp100 specific T cell responses after saline and L-Tyrosine vaccines, detected by IFN-γ positive CD8 T cells. B) Endogenous OVA-I specific T cell responses after saline and L-Tyrosine vaccines, detected by OVA-I dextramer H2-Kb positive CD8 T cells. Boosters were given as indicated times, with same dose and formulations as for priming. n = 3 - 5 mice per group.

Article Snippet: The synthetic, high-affinity H-2D b -restricted heteroclitic mouse gp100 25–33 peptide (KVPRNQDWL) and H-2K b –restricted chicken OVA-I 257–264 peptide (SIINFEKL) were purchased from Peptides International (Louisville, KY) at a purity >95%.

Techniques: Saline, Vaccines