Enhanced expansion of tumor-infiltrating lymphocytes for adoptive cell therapy

A method for treating a tumor in a subject, comprising the steps of:

• (a) expanding tumor-infiltrating lymphocytes (TILs) from a tumor fragment from the subject, the expanding comprising culturing the TILs in a two-stage process:
• i. culturing tumor fragments from the subject in a first cell culture medium comprising IL-2 and a 4-1BB agonist antibody;
• ii. removing at least a plurality of the TILs;
• and
• iii. expanding the plurality of TILs in a second cell culture medium comprising IL-2 and CD3 antibody to produce an expanded population of TILs;
• (b) treating the subject with nonmyleoablative lymphodepleting chemotherapy;
• and
• (c) administering the expanded population of TILs to the subject.