부산대학교 도서관

Cutaneous squamous cell carcinoma (cSCC) is one of the commonest cancers worldwide with the capacity to metastasize. Immune evasion by the tumour prevents its elimination and allows it to progress, including enabling the cancer to spread to local lymph nodes and distant organs. Targeting this dysfunctional immunity is a key strategy for the treatment of cSCC and anti-PD-1 checkpoint inhibition is now the first-line therapy for advanced and metastatic cSCC. However, complete responses to anti-PD-1 are seen in < 15% of patients and therefore it is vital that other strategies for enhancing anticancer immunity in cSCC are explored. Given the expense of clinical trials, it is essential to have good preclinical data to target cancer drug development appropriately, and this can be challenging in the investigation of cancer immunotherapy, as these treatments rely upon an intact human immune–tumour interface. We have developed a tumour slice culture (TSC) system to investigate the mechanism and efficacy of immune-modulating anticancer therapy in cSCC. This patient-derived explant model involves taking tissue slices from freshly excised tumours and culturing the slices under different experimental conditions, while maintaining the tumour microarchitecture of each slice. We cultured > 50 tumours using the TSC platform, including 35 cSCCs. Immunofluorescent confocal microscopy of anti-CD4 demonstrates penetration of antibody throughout the tissue slice. The mean (SD) viability of lymphocytes (T and B cells) determined by flow cytometry after 72 h of culture was 90.3% (7.89%; n = 31). Treatment of slices with OKT3 anti-CD3 antibody for 72 h induces a dose–response increase in T-cell Ki67 expression (2.87% unstimulated vs. 13% OKT3 100 ng mL–1; P < 0.001) and CD8+ T-cell granzyme B (47.6% unstimulated vs. 63.7% OKT3 ng mL–1; P = 0.064), as detected by flow cytometry (n = 7). Enzyme-linked immunosorbent assay of the culture supernatant showed that interferon-γ concentrations increased from <9.6 pg mL–1 (unstimulated) to 131.1 pg mL–1 [OKT3 1000 ng mL–1 (n = 6, P < 0.05]. To our knowledge the TSC system has not previously been used in cSCC and represents a promising technique to investigate new potential immunotherapies in this cancer. [ABSTRACT FROM AUTHOR]