Journal: Science Advances

Article Title: Immunogenic cryptic peptides dominate the antigenic landscape of ovarian cancer

doi: 10.1126/sciadv.ads7405

Figure Lengend Snippet: ( A ) A schematic of the workflow used for in vitro stimulation of patient T cells. Patient samples were stimulated with either a negative control peptide (10 μg/ml), the CEF peptide pool (5 μg/ml, positive control), or individual cryptic peptides (10 μg/ml) for 14 days, followed by another round of stimulation on day 15, and cells were expanded for another 8 days. On the day of the assay, the T cells were restimulated with peptide-pulsed autologous tumor cells and analyzed by flow cytometry. CEF is a pool of 32 class I peptides from cytomegalovirus, Epstein-Barr virus, and influenza virus. ( B ) Flow plots of intracellular IFN-γ expression in CD8 T cells from patient sample T2. ( C ) The percentage of CD8 T cells expressing either 4-1BB, TNFα, or IFN-γ following peptide stimulation in four OC samples. The colored dashed lines depict the baseline expression of 4-1BB, TNFα, and IFN-γ in negative peptide (neg pep)–stimulated T cells. ( D ) Frequencies of tetramer-positive CD8 T cell populations in samples T1 and T3 after a 19-day culture. HLA-A02:01–restricted peptides from HIV (gag) and cytomegalovirus (CMV; pp65) were used to generate tetramers that served as negative and positive controls, respectively. rhGM-CSF, recombinant human granulocyte-macrophage colony stimulating factor; LPS, lipopolysaccharide; PE, phycoerythrin.

Article Snippet: As positive control, pool of 32 class I peptides from cytomegalovirus, Epstein-Barr virus, and influenza virus (CEF; STEMCELL Technologies, #100-0675) was used.

Techniques: In Vitro, Negative Control, Positive Control, Flow Cytometry, Virus, Expressing, Recombinant