Growth of Tumor Derived Activated T-Cells for the Treatment of Cancer

Abstract

This report describes the production of Tumor Derived Activated Cell cultures (TDAC, also called tumor infiltrating lymphocytes) from patient tumor biopsies and our preliminary experience growing these cells to therapeutic levels using artificial capillary bioreactor cultures. TDAC were successfully grown in medium containing Interleukin 2 from 80% of the 113 tumor biopsies tested. There was no significant difference in success (growth to 1 x 10(9) cells) comparing primary and metastatic tumors. Many of the tumors were shipped to the laboratory from distant sites. Success rate did decrease with the length of time for tumor transport. Interleukin 4 was beneficial in the development of 1 of 4 TDAC cultures which did not grow with IL-2 only. Seventy-seven bioreactor cultures were initiated for 31 patients. On the average, 1.9 x 10(9) TDAC were inoculated per bioreactor; 3.3 x 10(10) were harvested in 22 days. Twelve liters of medium were required per 1 x 10(10) TDAC produced. TDAC cultures contained T cells with variable ratios of CD4 to CD8 cells. Secreted granulocyte monocyte colony stimulating factor, interferon gamma and tumor necrosis factor were measured in the bioreactor cartridge conditioned medium. Twenty three patients were evaluated. Partial responses were observed in 4 patients including a dramatic remission of scalp nodules in a patient with renal cancer. Results showed that therapeutic amounts of TDAC cells may be produced in a reasonable and cost effective manner using artificial capillary bioreactor cultures.

Characterization of Human Tumor-Infiltrating Lymphocytes Expanded in Hollow-Fiber Bioreactors for Immunotherapy of Cancer

“…This report is in follow-up to an earlier report encompassing the years 1991-1995 during which time TIL were initiated in T-flasks but were expanded using only 3L-gas permeable culture bags. 10 The overall success rate for initiation of TIL in this series is 76%, which is comparable to the success rate of 71% in our previous series, and comparable to the success rates reported by ourselves and other investigators 7,9,10,14,15 including another recent report using bioreactors. 25 We used the TIL expanded by bioreactor for a 111 Indium labeled TIL study that showed specific trafficking to tumors in patients with melanoma, renal cell carcinoma, and colon cancer.…”

“…However, this technique required significantly longer periods of cultivation time to achieve therapeutic levels, as compared to the single cell technique, approximately 78 days on the average. 7 Also, like the single cell concept, expansion of the TIL involves significant technical time and incubator space as a large number of flasks and/or bags are required to achieve a therapeutic dose.…”

“…3,4 There are many strategies and protocols for TIL isolation, expansion and treatment. [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21] In clinical pilot studies, durable responses have been reported in patients with metastatic cancer who were treated with infusions of TIL and IL-2, especially in patients with renal cell cancer and melanoma. [11][12][13][14][15][16][17][18][19][20][21] Unfortunately, the only randomized trial that attempted to ascertain whether IL-2 1 TIL was superior to IL-2 failed because of difficulties in growing TIL for the patients who were supposed to receive cell therapy.…”

“…Although much of the design and development of bioreactors has been geared towards production of antibodies by hybridoma cells or production of recombinant proteins by unicellular organisms, bioreactor technology has also been applied to the expansion of TIL for patient therapy. 5,[7][8][9] Hollow-fiber bioreactors, such as those investigated by us, create a microenvironment that can be specially suited for TIL expansion. Basically, TIL are seeded into extracapillary space (ECS) outside of the hollow-fiber capillaries thus allowing expansion of TIL in the ECS in the continuous presence of medium containing IL-2 and essential nutrients while simultaneously allowing removal of cell waste products.…”

New Miniaturized Hollow-Fiber Bioreactor for in Vivo Like Cell Culture, Cell Expansion, and Production of Cell-Derived Products

“…An alternative cell culture technology is the hollow-fiber bioreactor, first discribed by Knazek et al (1). With this technology it is, in principle, possible to overcome some limitations of monolayer cultures, to realize increased cell densities and concentrations of secreted effector molecules (2)(3)(4), and to permit a continuous cell cultivation process approaching tissue-like cell densities (5). Most hollow-fiber bioreactors are similar in design and size to modules used in hemodialysis of uremic patients (6).…”