To perform the cytotoxicity study of Pembrolizumab, we preliminary tested the 1:1 and 1:5 ratio of tumor cells:PMBCs, and as the 1:1 ratio achieved almost 15% inhibition of cell viability, in agreement with our previous data [8], in order to amplify the impact of EVs on the response to anti-PD1 we chose to conduct the assay by using the 1:5 ratio of tumor cells:PMBCs. Here, PDCD1 is linked to neoplasm.