This study evaluated the efficacy and safety of autologous tumor-infiltrating lymphocytes (TIL) in patients with recurrent or refractory ovarian cancer (OVCA), colorectal cancer (CRC), and pancreatic ductal adenocarcinoma (PDAC). The trial involved 16 patients who received TIL therapy, manufactured with IL-2 and agonistic stimulation of CD3 and 4-1BB. The primary endpoint was the objective response rate (ORR) at 12 weeks, while secondary endpoints included disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and safety. No objective responses were observed, with an ORR of 0%. However, 62.5% of patients achieved stable disease (SD), indicating some clinical benefit. The median PFS was 2.53 months, and the median OS was 18.86 months. Grade 3 or higher toxicities were observed in 87.5% of patients. TIL products showed effector memory cells with high CD39 expression and low checkpoint marker expression, suggesting a functionally active TIL population. The study demonstrated the feasibility and safety of TIL therapy in these cancers, with no new safety signals. While no objective responses were observed, the presence of SD and prolonged SD in one PDAC patient suggests potential immunological activity. Further research is needed to enhance TIL efficacy through improved manufacturing and genetic modifications.This study evaluated the efficacy and safety of autologous tumor-infiltrating lymphocytes (TIL) in patients with recurrent or refractory ovarian cancer (OVCA), colorectal cancer (CRC), and pancreatic ductal adenocarcinoma (PDAC). The trial involved 16 patients who received TIL therapy, manufactured with IL-2 and agonistic stimulation of CD3 and 4-1BB. The primary endpoint was the objective response rate (ORR) at 12 weeks, while secondary endpoints included disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and safety. No objective responses were observed, with an ORR of 0%. However, 62.5% of patients achieved stable disease (SD), indicating some clinical benefit. The median PFS was 2.53 months, and the median OS was 18.86 months. Grade 3 or higher toxicities were observed in 87.5% of patients. TIL products showed effector memory cells with high CD39 expression and low checkpoint marker expression, suggesting a functionally active TIL population. The study demonstrated the feasibility and safety of TIL therapy in these cancers, with no new safety signals. While no objective responses were observed, the presence of SD and prolonged SD in one PDAC patient suggests potential immunological activity. Further research is needed to enhance TIL efficacy through improved manufacturing and genetic modifications.