Figure S1 from Vaccination with Designed Neopeptides Induces Intratumoral, Cross-reactive CD4<sup>+</sup> T-cell Responses in Glioblastoma

Schematic outline of the personalized peptide vaccination. (A) Timeline of clinical events for a patient with glioblastoma in whom we applied a highly personalized peptide vaccination. I. Vaccine peptide design. Surgically resected primary glioblastoma and PBMCs were collected. Somatic mutants and over-/highly expressed genes in glioblastoma were identified as TSAs and TAAs by WES and RNA-seq, and the HLA genotype were identified with PBMCs by DNA sequencing. Potential vaccine peptides were predicted and selected from TSAs and TAAs based on HLA binding prediction. Peptide cocktails containing ~25 peptides for HLA class I or II molecules were prepared for vaccination. II. Peptide cocktail vaccination and testing. The patient was s.c. vaccinated with the peptide cocktails, and Imiquimod or XS15 + MontanideTM were used as the adjuvant. Meanwhile, the patient received treatment with pembrolizumab and bevacizumab. No dexamethasone was given throughout the periods of vaccination. PBMCs were collected after vaccination at the indicated time points and tested with vaccine peptides to examine the effect of immunotherapy. Surgically resected recurrent glioblastoma was collected after four vaccinations, and tumor-infiltrating lymphocytes (TILs) were isolated and tested with vaccine peptides. (B) HLA genotypes of the patient with glioblastoma, including HLA-A, HLA-B, HLA-DR, HLA-DP, and HLA-DQ alleles. n.a. indicates not available.