cd8 t cell enrichment  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: Immunodominance Hierarchy of Endogenous BBN963 Bladder Cancer Antigens Remains Stable Under Anti-PD1 and Anti-CTLA4 Immunotherapy

doi: 10.64898/2026.05.20.726664

Figure Lengend Snippet: Three BBN963 tumors were analyzed by whole exome and transcriptome sequencing. Data were analyzed using the Landscape of Effective Tumor antigen Software (LENS) to predict tumor antigens derived from a set of genomic sources (SNVs, indels, fusions, splice variants, viruses, endogenous retroviruses, and cancer/testis antigens). The set of predicted tumor antigen peptide epitopes was evaluated using ACE Configurator for ELISpot (ACE) to construct peptide matrices for large scale ELISpot testing. IFN-γ production measured by ELISpot was used to determine relative immunodominance of MHC Class I restricted epitopes recognized by CD8+ T cells.

Article Snippet: A kit was used for CD8+ T cell enrichment (Miltenyi Biotec, Cat No 130-104-075) via magnetic column, with calculations and procedures done according to the manufacturer instructions.

Techniques: Sequencing, Software, Derivative Assay, Enzyme-linked Immunospot, Construct

Journal: bioRxiv

Article Title: Immunodominance Hierarchy of Endogenous BBN963 Bladder Cancer Antigens Remains Stable Under Anti-PD1 and Anti-CTLA4 Immunotherapy

doi: 10.64898/2026.05.20.726664

Figure Lengend Snippet: 6-week-old C57BL/6 female mice were inoculated with 3 x 10 6 BBN963 cells, sacrificed at 10 days post tumor inoculation, and assessed for tumor antigen-specific CD8+ T cell responses via IFN-g ELISpot in pooled animal studies n=15 and individual animal studies (biological replicates n=9). (A) IFN-g spot count of matrixed peptide pools in pooled animal studies. Peptide pools containing positive antigens are shown in magenta and pools that are negative shown in light blue. (B) Single peptide validation of tumor antigens via ELISpot. Tumor antigens are mapped to genomic source and predicted MHC restriction (H2-Kb and H2-Db). (C) Immunodominance hierarchy of Tumor Antigens across immunogenic antigens with peptide biophysical similarity cladogram displaying ELISpot response to peptides with same core sequences. (D) Validation of immunodominance hierarchy of immunoprevalent antigens in single animal studies.

Article Snippet: A kit was used for CD8+ T cell enrichment (Miltenyi Biotec, Cat No 130-104-075) via magnetic column, with calculations and procedures done according to the manufacturer instructions.

Techniques: Enzyme-linked Immunospot, Biomarker Discovery

Journal: bioRxiv

Article Title: Immunodominance Hierarchy of Endogenous BBN963 Bladder Cancer Antigens Remains Stable Under Anti-PD1 and Anti-CTLA4 Immunotherapy

doi: 10.64898/2026.05.20.726664

Figure Lengend Snippet: (A) Experimental design: 6-week-old C57BL/6 female mice were inoculated with 3 x 10 6 BBN963 cells. Assessment of antigen-specific CD8 + T cell responses was done as outlined in (n=10 per treatment arm). (B) 145 peptide matrixed IFN-γ ELISpot results of anti-PD1 treated tumor-bearing animals as deconvoluted by ACE. (C) Single peptide validation of anti-PD1 treated animals showing the genomic source of each antigen. (D) 145 peptide matrixed IFN-γ ELISpot results of anti-CTLA4 treated tumor bearing animals as deconvoluted by ACE. (E) Single peptide validation of anti-CTLA4 treated animals showing the genomic source of each antigen.

Article Snippet: A kit was used for CD8+ T cell enrichment (Miltenyi Biotec, Cat No 130-104-075) via magnetic column, with calculations and procedures done according to the manufacturer instructions.

Techniques: Enzyme-linked Immunospot, Biomarker Discovery