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: Vaccines

Article Title: Murine Dendritic Cells Grown in Serum-Free Culture Show Potent Therapeutic Activity when Loaded with Novel Th Epitopes in an Orthotopic Model of HER2 pos Breast Cancer

doi: 10.3390/vaccines9091037

Figure Lengend Snippet: Murine DCs cultured in a serum-free medium can prime in vivo against both MHC class I and class II-restricted antigens. ( A ) Schema of sensitization protocol. Balb/c mice were vaccinated subcutaneously in the flank on day 0 with 1 × 10 6 SFM-DC loaded with either OVA (ISQAVHAAHAEINEAGR) MHC class II peptide, rat ErbB2/HER2 (TYVPANASL) MHC class I peptide or left unpulsed. Mice received an identical booster on day 21, and on day 35 mice were sacrificed and immune splenocytes subjected to analysis. ( B ) IFN-γ ELISA analysis of 24 h culture supernatants from immune splenocyte (three mice per treatment group) stimulated with various recall antigens. ( C ) IFN-γ ELISPOT analysis from stimulated splenocytes from individual mice. ( D ) Digital images of ELISPOT wells from representative mouse (#6) performed in either commercial ELISPOT culture medium (CEM; upper images) or base RPMI medium (lower images). ( E ) Enumeration of IFN-γ spots from splenocytes of vaccinated mice stimulated in base RPMI medium without added serum. Error bars for all panels indicate SEM.

Article Snippet: Rat ErbB2/HER2 cytotoxic T cell (CTL) epitope (TYVPANASL) was synthesized by Bachem (Torrance, CA, USA) and ovalbumin helper T (Th) epitope ISQAVHAAHAEINEAGR was purchased from InvivoGen (San Diego, CA, USA).

Techniques: Cell Culture, In Vivo, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: Vaccines

Article Title: Murine Dendritic Cells Grown in Serum-Free Culture Show Potent Therapeutic Activity when Loaded with Novel Th Epitopes in an Orthotopic Model of HER2 pos Breast Cancer

doi: 10.3390/vaccines9091037

Figure Lengend Snippet: Comparison of efficacy of peptide-pulsed DCs cultured in serum-free versus serum-replete medium in a therapy model for HER2 pos breast cancer. ( A ) Schema of therapy protocol. On day 0, fifteen female Balb/c mice received 1 × 10 6 orthotopically-implanted TUBO breast carcinoma cells. On day 10 mice were divided into three groups (five mice each) and vaccinated subcutaneously in the flank with rat ErbB2/HER2 class I peptide (TYVPANASL) pulsed either onto DCs cultured in X-Vivo 20 (SFM-DC) or in RPMI supplemented with 10% fetal calf serum (FCS-DC). Alternatively, mice were left untreated (No Rx). ( B ) growth curves of tumors over the course of 28 days. ( C ) Average tumor size comparison on day 28 with statistical analysis. ( D ) Kaplan–Meyer survival plot where vaccinated mice were followed for an additional 30 days. Failure to survive was defined as tumor size exceeding size of 200 mm 2 .

Article Snippet: Rat ErbB2/HER2 cytotoxic T cell (CTL) epitope (TYVPANASL) was synthesized by Bachem (Torrance, CA, USA) and ovalbumin helper T (Th) epitope ISQAVHAAHAEINEAGR was purchased from InvivoGen (San Diego, CA, USA).

Techniques: Comparison, Cell Culture

Journal: Vaccines

Article Title: Murine Dendritic Cells Grown in Serum-Free Culture Show Potent Therapeutic Activity when Loaded with Novel Th Epitopes in an Orthotopic Model of HER2 pos Breast Cancer

doi: 10.3390/vaccines9091037

Figure Lengend Snippet: Identification of Helper T cell epitopes on rat ErbB2/HER2. ( A ) Schema of peptide vaccination protocol. On day 0 female Balb/c mice were vaccinated subcutaneously (s.q.) with pools of 5–6 peptides (30 µg each) based on the rat ErbB2/HER2 extracellular domain sequence and emulsified in Freund’s Complete Adjuvant. On day 21 mice received an identical booster immunization in Freund’s Incomplete adjuvant. Mice were bled 12–14 days after each vaccination (about day 14 and 35) to obtain immune sera. ( B ) ELISA analysis for IgG production against individual pool peptides. Four peptides (indicated by arrows) were selected for further study: p9; QEVQGYMLIAHNQVKRVPLQR, P17; GNPQLCYQDMVLWKDVFRKN, p35; PCARVCYGLGMEHLRGARAI and p48; LIHRNAHLCFVHTVPWDQLF). ( C ) Schema for therapeutic DC vaccination with novel putative Th peptides. Female Balb/c mice (five per treatment group) received 1 × 10 6 TUBO breast carcinoma cells implanted orthotopically on day 0. On days 14 and 21 mice received subcutaneous flank vaccination with 1 × 10 6 SFM DCs pulsed with putative Th peptides p9, p17, p35 and p48. Other groups received DCs pulsed with rat ErbB2/HER2 CTL peptide TYVPANASL (Class I), or unpulsed DC. A final group of tumor-bearing mice was left untreated (No Rx). Mice were monitored for tumor size twice weekly and were sacrificed on day 29. ( D ) Tumor growth curves for mice receiving various indicated SFM-DC vaccines. ( E ) Splenocytes from sacrificed mice vaccinated with SFM-DC pulsed with either p9, p17, p35 or p48 were cultured in the presence of various recall peptides and 24 h supernatants analyzed for IFN-γ via ELISA analysis. Error bars indicate SEM.

Article Snippet: Rat ErbB2/HER2 cytotoxic T cell (CTL) epitope (TYVPANASL) was synthesized by Bachem (Torrance, CA, USA) and ovalbumin helper T (Th) epitope ISQAVHAAHAEINEAGR was purchased from InvivoGen (San Diego, CA, USA).

Techniques: Sequencing, Adjuvant, Enzyme-linked Immunosorbent Assay, Vaccines, Cell Culture

Journal: Current Oncology

Article Title: Non-Mutated Nucleophosmin 1 Is Recognized by the CD8+ T Lymphocytes of an AML Patient after the Transplantation of Hematopoietic Stem Cells from an HLA-Haploidentical Donor

doi: 10.3390/curroncol29050239

Figure Lengend Snippet: Nucleophosmin-specific T cell responses in an AML patient after HSCT. ( A ) Frequency of spot-forming units (SFU) specific for NPM1 wt detected by ELISPOT-IFNγ and result of quantification of NPM1c + transcripts in bone marrow. Time of DLI is denoted by arrows. ( B ) The phenotype of NPM1 wt -specific T cells at month +3 producing IFNγ determined by IC FACS. ( C ) Frequency of T cells specific for CTL epitopes 93-EITPPVVLR, 142-RSAPGGGSK, 194-KSIRDTPAKNAQK, 242-SSVEDIKAK, and 259-GSLPKVEAK determined at month +5. Statistical analysis was performed using the Mann–Whitney test ( A , C ) **** p < 0.0001, *** p < 0.001, * p < 0.05.

Article Snippet: Five candidate NPM1 wt CTL epitopes, 93-EITPPVVLR, 142-RSAPGGGSK, 194-KSIRDTPAKNAQK, 242-SSVEDIKAK, and 259-GSLPKVEAK, restricted by HLA-A*11:01, were predicted using the Syfpeithi [ ] and NetCTLpan [ ] algorithms and synthesized by JPT Peptide Technologies GmbH.

Techniques: Enzyme-linked Immunospot, MANN-WHITNEY

Journal: Frontiers in Veterinary Science

Article Title: Epitopes screening and vaccine molecular design of SADS-CoV based on immunoinformatics

doi: 10.3389/fvets.2022.1080927

Figure Lengend Snippet: Descriptive workflow for the epitope-based candidate vaccine prediction.

Article Snippet: MHC-I combined CTL epitope prediction by NetMHCpan4.1 server ( https://services.healthtech.dtu.dk/service.php?NetMHCpan-4.1 ) and IEDB server ( http://tools.iedb.org/mhci/ ).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Epitopes screening and vaccine molecular design of SADS-CoV based on immunoinformatics

doi: 10.3389/fvets.2022.1080927

Figure Lengend Snippet: SADS-CoV multi-epitope vaccine kit.

Article Snippet: MHC-I combined CTL epitope prediction by NetMHCpan4.1 server ( https://services.healthtech.dtu.dk/service.php?NetMHCpan-4.1 ) and IEDB server ( http://tools.iedb.org/mhci/ ).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Epitopes screening and vaccine molecular design of SADS-CoV based on immunoinformatics

doi: 10.3389/fvets.2022.1080927

Figure Lengend Snippet: Modeling of epitope vaccine tertiary structure. (A–E) Omega, Phi, Distance, Theta and Contact parameters of 2D structures; (F) Tertiary structure of candidate vaccine, Red: β-Defensin I molecular adjuvant; Yellow: CTL epitopes; Purple: Th epitopes; Green: B cell epitopes; TM-score: 0. 376.

Article Snippet: MHC-I combined CTL epitope prediction by NetMHCpan4.1 server ( https://services.healthtech.dtu.dk/service.php?NetMHCpan-4.1 ) and IEDB server ( http://tools.iedb.org/mhci/ ).

Techniques: Adjuvant

Journal: Frontiers in Veterinary Science

Article Title: Epitopes screening and vaccine molecular design of SADS-CoV based on immunoinformatics

doi: 10.3389/fvets.2022.1080927

Figure Lengend Snippet: Validation of tertiary structure of epitope vaccine. (A) Ramachandran plots to validate the tertiary structure; (B) For comparison of tertiary protein structure, the crude model and refined model are green and red, respectively; (C) The Ramachandran mapping validates the optimized tertiary structure; (D) The Z fraction of the ProSA SEB diagram of the refined model is −3.34; (E) The quality of the local model is shown by plotting the energy as a function of the position of the amino acid sequence.

Article Snippet: MHC-I combined CTL epitope prediction by NetMHCpan4.1 server ( https://services.healthtech.dtu.dk/service.php?NetMHCpan-4.1 ) and IEDB server ( http://tools.iedb.org/mhci/ ).

Techniques: Biomarker Discovery, Comparison, Sequencing

Journal: Frontiers in Veterinary Science

Article Title: Epitopes screening and vaccine molecular design of SADS-CoV based on immunoinformatics

doi: 10.3389/fvets.2022.1080927

Figure Lengend Snippet: Complexes of candidate vaccine protein docking with human TLR3 receptor. (A) Epitope vaccine structure is shown in blue and TLR3 in green; (B) Residue of interaction between docking TLR3 and epitope vaccine; (C) Local magnification highlights amino acid residues; (D) LigPLot+ determined two-dimensional interaction diagram.

Article Snippet: MHC-I combined CTL epitope prediction by NetMHCpan4.1 server ( https://services.healthtech.dtu.dk/service.php?NetMHCpan-4.1 ) and IEDB server ( http://tools.iedb.org/mhci/ ).

Techniques: Residue