mer neoantigen peptide  (MedChemExpress)

Journal: Signal Transduction and Targeted Therapy

Article Title: Oncolytic adenovirus delivery of neoantigens sensitizes low-mutation tumors to anti-PD-1 therapy and prevents metastasis

doi: 10.1038/s41392-025-02511-5

Figure Lengend Snippet: Neoantigen identification and delivery via oncolytic adenovirus vectors. a Schematic representation of neoantigen identification combining WES and MS. This figure was created via resources from BioRender ( www.biorender.com ). b Design of the neoantigen delivery system. The expression cassette of neoantigen cDNA sequences corresponding to the 29-mer long peptides covering the mutated epitopes was inserted into the E3 region of the survivin promoter (SVP)-regulated oncolytic adenovirus vector Ad5SVPF11 to generate a novel OAV, AdSVP-NAg. C57BL/6 mice inoculated with B16F10 melanoma, MC38 colon carcinoma, or Hep53.4 hepatocellular carcinoma cells were treated with PBS, vector (Ad5SVPF11), SLP, vector combined with SLP, or AdSVP-NAg. Tumor growth curves ( c ) ( n = 6 mice per group), representative images ( d ) of the IFN-γ ELISpot of neoantigen-specific T cells from the spleen in each group and immunofluorescence images ( e ) of tumor tissues. The data are shown as the means ± SDs ( c ) and are representative of two independent experiments ( c – e ). Significance was calculated by two-way ANOVA ( c ). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The formulation of the SLP vaccine was adapted from previous studies, with each dose containing 20 μg of each 29-mer neoantigen peptide in Supplementary Table and 50 μg of poly I:C (MedChemExpress, USA), dissolved in 200 μL of a 10% DMSO/90% PBS solution.

Techniques: Expressing, Plasmid Preparation, Enzyme-linked Immunospot, Immunofluorescence

Journal: Nature

Article Title: MHC-II neoantigens shape tumor immunity and response to immunotherapy

doi: 10.1038/s41586-019-1671-8

Figure Lengend Snippet: (a-b) T3 MHC-II neoantigen predictions for all expressed mutations were made using hmMHC (a) and netMHCII-2.3 (b) (netMHCIIpan-3.2 predictions yield very similar results). The predictions are shown as −10 log odds predictor value or logIC50 (smaller values indicate higher likelihood of being presented by I-A b ) and expression level (FPKM). Strong binders are defined as mutations residing in the 2 nd percentile of I-A b binding predictions for random natural peptides for each algorithm (−10logOdds ≤ 26.21 or IC50 ≤ 343.8 nM). The N710Y mutation in Itgb1 met the strong binder threshold in the hmMHC predictions but not in the netMHCII-2.3 predictions. Red dots indicate all mutations that were screened for CD4 + T cell reactivity. Green line denotes high expression cutoff (FPKM=89.1). Blue line indicates strong binder cut off for each algorithm. (c) Two million T3 sarcoma cells were injected subcutaneously into syngeneic mice and CD4 + TIL was isolate on day 12. IFNγ ELISPOT was performed using naïve splenocytes pulsed with 2 μg mL −1 of the indicated peptides. Data is shown as average of three independent experiments ± SEM. (d) Gating strategy for pI-A b tetramer staining of whole TIL. (e) Quantification of mITGB1-tetramer and CLIP-tetramer staining of CD4 + T cells from whole T3 TIL 12 days post-transplant. Data is shown as average percent tetramer-positive cells of CD4 + cells ± SEM of 3 independent experiments. (f) Syngeneic 129S6 mice were injected subcutaneously with 2x10 6 T3 sarcoma cells and TIL-derived CD4 + T cells were harvested 12 days post transplant. CD4 + T cells were stimulated with naïve splenocytes pulsed with 2 μg/mL OVA 323-339 control or mITGB1 697-724 peptide for a flow-based multi-cytokine array. Representative data from one of two independent experiments using pools of 5 tumors each is shown as average of technical triplicate wells from 3 pooled tumors.

Article Snippet: All 27-mer peptides used for neoantigen screening ( ) were purchased from Peptide 2.0 and HPLC purified to >95% purity.

Techniques: Expressing, Binding Assay, Mutagenesis, Injection, Enzyme-linked Immunospot, Staining, Derivative Assay

Journal: Nature

Article Title: MHC-II neoantigens shape tumor immunity and response to immunotherapy

doi: 10.1038/s41586-019-1671-8

Figure Lengend Snippet: (a) Comparison of total number of expressed missense mutations between 10 different MCA-induced sarcomas and KP9025. Mutations were defined by WES and RNAseq and mutational load is shown on a per cell basis. (b) Comparison of predicted neoantigen MHC-I affinity values between KP9025 and MCA-induced sarcoma F244 for H-2D b (top) and H-2K b (bottom). KP9025 were not predicted to express any MHC-I neoantigens. (c) Rag2 −/− mice were subcutaneously injected with 1x10 6 KP.mLAMA4, KP.mITGB1, KP.mLAMA4.mITGB1 or KP.mSB2.SIINFEKL. Representative data from one of two independent experiments is presented as tumor diameter of individual mice (n=5 KP.mLAMA4, KP.mITGB1 and KP.mLAMA4.mITGB1 and n=3 KP.mSB2.SIINFEKL mice per group per experiment) (d) WT syngeneic 129S4 mice were injected subcutaneously with 1x10 6 KP.mLAMA4, KP.mITGB1 or KP.mLAMA4.mITGB1 and treated with αPD-1 (top) or αCTLA single agent ICT (bottom) on days 3, 6, and 9 post transplant. Representative data from one of three independent experiments is shown as tumor diameter from individual mice (n=5 in all groups per experiment). (e) Survival curves from all experiments described in (d) and Figure 2 e (n=15 in all groups).

Article Snippet: All 27-mer peptides used for neoantigen screening ( ) were purchased from Peptide 2.0 and HPLC purified to >95% purity.

Techniques: Injection