Fig. 6: Identification of neoantigen and HPV-reactive cells in patients after anti-OX40. | Nature Communications

Fig. 6: Identification of neoantigen and HPV-reactive cells in patients after anti-OX40.

From: Neoadjuvant anti-OX40 (MEDI6469) therapy in patients with head and neck squamous cell carcinoma activates and expands antigen-specific tumor-infiltrating T cells

Fig. 6

Expanded DP CD8+ T cells from HNOX18 were screened for reactivity against HPV16, and neoantigens were predicted for HNOX04 and HNOX07. Peptide Pools were screened for reactivity by IFN-γ -ELISpot analysis. a Expanded DP CD8+ T cells from HNOX18 were screened with autologous PBMC transfected by electroporation with RNA encoding HPV16 E6 and E7 proteins. Anti-CD3 is a positive control, water the negative (MOCK) control. Prior to IFN-γ ELISpot development (right), cells were harvested and expression of 4-1BB and CD25 was assessed by flow cytometry (left). b Summary of HPV-specific spot-forming cells (SFC) in CD8+ TIL subsets. c HPV16 E6 and HPV16 E7 reactive CD8+ T cells were sorted based on the expression of 4-1BB and CD25. 4-1BB-CD25− and 4-1BB+ CD25+ cells were analyzed by TCRβ-sequencing and the frequency of the top 6 (E6) and top 7 (E7) clones are depicted. d Expanded DP CD8+ T cells from patient HNOX04 were screened with the addition of predicted neoantigens. Shown is the response to peptide 5, anti-CD3 as positive, DMSO as the negative control. e Summary of spot-forming cells (SFC) in CD8+ TIL subsets from HNOX04. f Expanded DP T cells from HNOX07 were screened with the addition of predicted neoantigens. Shown is the response to peptide 21, anti-CD3 as positive, DMSO as the negative control. g Summary of SFC in CD8+ TIL subsets from HNOX07. Red depicts the peptide response, gray is the anti-CD3 control, black the MOCK/DMSO control. TMG tandem minigene. Source data are provided as Source Data file.

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