Journal: Molecular Cancer Therapeutics

Article Title: Novel Chimeric CTLA-4 B-cell Epitope Peptide Vaccines Demonstrate Effective Antitumor Immunity with/without PD1/PDL1 Blockade in Multiple Syngeneic Murine Models of Breast and Colorectal Cancers

doi: 10.1158/1535-7163.MCT-24-0908

Figure Lengend Snippet: Combination CTLA-4 with PDL1 peptide vaccine in CT26-BALB/c tumor model. A, Six- to eight-week-old BALB/c mice were vaccinated with combination peptides as designed [MVF-PDL1 (130) with MVF-CTLA-4 (130)] for 3-week interval. A measure of 0.1 mg of each peptide cancer vaccine mixed with ISA 720 (1:1) used per mouse. Mice were boosted with the designed doses for every 3-week intervals. Blood was collected weekly for monitoring antibody titers. After 2 weeks of the third time immunization (3Y), mice were challenged with 2 × 10 5 per mouse D2F2 tumor cells or 5 × 10 5 per mouse 4T1 tumor cells. After tumor challenge, in the positive control group, we treated the mice with anti-mouse antibodies 10F.9G2 plus 9H10 twice a week for up to 3 weeks, and the negative control group was treated with PBS. Tumor volume was calculated as follows: Tumor volume (LWW) = (length × width × width)/2. B, Immunogenicity of MVF-CTLA-4 plus MVF-PDL1 (130) peptides immunized BALB/c mice. Mice bleeds were collected weekly except the first collection was 3 weeks after primary immunization, and ELISA was used to detect antibody titers in sera; C, Antibody isotypes from mice bleed immunized with MVF-CTLA-4 (130) plus MVF-PDL1 (130) combination peptide vaccine. D, Tumor burden (LWW) by days in BALB/c mice of each treatment group (10 mice/group) and tumor models of both D2F2 and 4T1 BALB/c mammary carcinoma, and two-way ANOVA was used to analyze the whole curves of tumor growth. Tumor burden (LWW) analysis in BALB/c mice of each treatment group and tumor models, and one-way ANOVA was used to analyze at each time point as indicated at days 12 and 14 for the D2F2 model and days 9 and 12 for the 4T1 model after tumor challenging. E, The log-rank (Mantel–Cox) test was used to compare the survival curves. ns, no significant; *, P < 0.05; **, P < 0.01, all vs. with the PBS control group.

Article Snippet: BALB/c mouse CT26 colon cancer cell line wild-type (WT; CT26 WT and CRL-2638, RRID: CVCL_7256) and 4T1 (CRL-2539, RRID: CVCL_0125) TNBC tumor cell lines were purchased in 2018 and 2020 from the ATCC.

Techniques: Positive Control, Negative Control, Immunopeptidomics, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecular Cancer Therapeutics

Article Title: Novel Chimeric CTLA-4 B-cell Epitope Peptide Vaccines Demonstrate Effective Antitumor Immunity with/without PD1/PDL1 Blockade in Multiple Syngeneic Murine Models of Breast and Colorectal Cancers

doi: 10.1158/1535-7163.MCT-24-0908

Figure Lengend Snippet: CTLA-4 peptide mimics therapeutic efficiency in syngeneic mouse model. A, The BALB/c mice of 6–8 weeks old were challenged with 1 × 10 5 CT26 tumor cells on day 0 (10 mice/group). Then the mice were received 200 μg/dose anti-CTLA-4 mAb (9H10) or CTLA-4 peptide mimics starting from day 1. The following injections are indicated in the schematic figure. PBS as a negative control and the 9H10 group serves as a positive control. The tumor volume and mice condition were monitored regularly. B, Tumor burden (LWW) by days in BALB/c mice of each treatment group (10 mice/group) and tumor models, and two-way ANOVA was used to analyze the whole curves of tumor growth. C, Tumor burden (LWW) analysis in BALB/c mice of each treatment group and tumor models, and one-way ANOVA was used to analyze at each time point as indicated. (9 mice in the PBS group on day 16 and one mouse were removed from PBS group on day 14 after tumor challenging). D, The log-rank (Mantel–Cox) test was used to compare the survival curves. ns, no significant; *, P < 0.05; **, P < 0.01, all vs. with the PBS control group.

Article Snippet: BALB/c mouse CT26 colon cancer cell line wild-type (WT; CT26 WT and CRL-2638, RRID: CVCL_7256) and 4T1 (CRL-2539, RRID: CVCL_0125) TNBC tumor cell lines were purchased in 2018 and 2020 from the ATCC.

Techniques: Negative Control, Positive Control, Control

Journal: Molecular Cancer Therapeutics

Article Title: Novel Chimeric CTLA-4 B-cell Epitope Peptide Vaccines Demonstrate Effective Antitumor Immunity with/without PD1/PDL1 Blockade in Multiple Syngeneic Murine Models of Breast and Colorectal Cancers

doi: 10.1158/1535-7163.MCT-24-0908

Figure Lengend Snippet: Combination CTLA-4 with PDL1 peptide vaccine in CT26-BALB/c tumor model. A, Six- to eight-week-old BALB/c mice (10 mice/group) were vaccinated with MVF peptides [MVF-CTLA-4 (59) or (130) with MVF-PDL1 (36) or (130)] as indicated for 3-week interval. Each peptide cancer vaccine 100 μg mixed with ISA 720 (1:1) was immunized per mouse. Mice were boosted with the designed doses, and bleeds were collected weekly for monitoring antibody titers. After 2 weeks of the third time immunization (3Y), mice were challenged with 1 × 10 5 CT26 tumor cells on day 0. The tumor growth checked daily, especially 7 days after challenging, and tumor size was measured using calipers. For the positive control group, we treated the mice with anti-mouse CTLA-4 antibody (clone 9H10) plus anti-mouse PDL1 antibody (10F.9G2) twice a week for up to 3 weeks, whereas the negative control group was treated with PBS. B, Immunogenicity of combination MVF-CTLA-4 with MVF-PDL1 peptides immunized BALB/c mice. Mice bleeds were collected weekly except the first collection was 3 weeks after primary immunization, and ELISA was used to detect antibody titers in sera; C, Tumor burden (LWW) by days in BALB/c mice of each treatment group (10 mice/group) and tumor models, and two-way ANOVA was used to analyze the whole curves of tumor growth. Tumor burden (LWW) analysis in BALB/c mice of each treatment group and tumor models, and one-way ANOVA was used to analyze at each time point as indicated at days 14 and 16 after tumor challenging. D, The log-rank (Mantel–Cox) test was used to compare the survival curves. ns, no significant; *, P < 0.05; **, P < 0.01, all vs. with the PBS control group.

Article Snippet: BALB/c mouse CT26 colon cancer cell line wild-type (WT; CT26 WT and CRL-2638, RRID: CVCL_7256) and 4T1 (CRL-2539, RRID: CVCL_0125) TNBC tumor cell lines were purchased in 2018 and 2020 from the ATCC.

Techniques: Positive Control, Negative Control, Immunopeptidomics, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecular Cancer Therapeutics

Article Title: Novel Chimeric CTLA-4 B-cell Epitope Peptide Vaccines Demonstrate Effective Antitumor Immunity with/without PD1/PDL1 Blockade in Multiple Syngeneic Murine Models of Breast and Colorectal Cancers

doi: 10.1158/1535-7163.MCT-24-0908

Figure Lengend Snippet: Combination CTLA-4 with PDL1 peptide vaccine in CT26-BALB/c tumor model. A, Six- to eight-week-old BALB/c mice were vaccinated with combination peptides as designed MVF-CTLA-4 (130) combination with PD1-Vaxx [MVF-PD1 (92)] for 3-week interval. A measure of 0.1 mg of each peptide cancer vaccine mixed with ISA 720 (1:1) used per mouse. Mice were boosted with the designed doses for every 3 weeks intervals. Blood was collected weekly for monitoring antibody titers. After 2 weeks of the third time immunization (3Y), mice were challenged with 1 × 10 5 CT26 per mouse, or 2 × 10 5 per mouse D2F2 tumor cells, or 5 × 10 5 per mouse 4T1 tumor cells. After tumor challenge, in the positive control group, we treated the mice with anti-mouse antibodies 9H10 plus 29F.1A12 twice a week for up to 3 weeks, and the negative control group was treated with PBS. Tumor volume was calculated as follows: Tumor volume (LWW) = (length × width × width)/2. B, Immunogenicity of MVF-CTLA-4 plus PD1-Vaxx peptides immunized BALB/c mice. Mice bleeds were collected weekly except the first collection was 3 weeks after primary immunization, and ELISA were used to detect antibody titers in sera. C, Antibody isotypes from mice bleed immunized with MVF-CTLA-4 (130) plus MVF-PD1 (92) combination peptide vaccine. D, Tumor burden (LWW) by days in BALB/c mice of each treatment group (10 mice/group) and tumor models of CT26, D2F2, and 4T1 BALB/c carcinoma, and two-way ANOVA was used to analyze the whole curves of tumor growth. Tumor burden (LWW) analysis in BALB/c mice of each treatment group and tumor models, and one-way ANOVA was used to analyze at each time point as indicated at days 9 and 12 for the CT26 model, days 12 and 14 for the D2F2 model, and days 9 and 12 for 4T1 model after tumor challenging. E, The log-rank (Mantel–Cox) test was used to compare the survival curves. ns, no significant; *, P < 0.05; **, P < 0.01, all vs. with the PBS control group.

Article Snippet: BALB/c mouse CT26 colon cancer cell line wild-type (WT; CT26 WT and CRL-2638, RRID: CVCL_7256) and 4T1 (CRL-2539, RRID: CVCL_0125) TNBC tumor cell lines were purchased in 2018 and 2020 from the ATCC.

Techniques: Positive Control, Negative Control, Immunopeptidomics, Enzyme-linked Immunosorbent Assay, Control