Activation and Expansion of Tumor-Derived Activated Cells for Therapeutic Use

Abstract

We obtained tumor specimens from patients with cancer in an effort to activate and expand the tumor-infiltrating lymphocytes for therapeutic use. With the use of finely minced tumor preparations from eight different tumor types, recombinant interleukin-2, and lymphokine-activated killer cell-conditioned medium, lymphocytes were expanded in vitro. After 4 weeks, the tumor cells were virtually absent from the cultures. At this point, the lymphocytes were termed "tumor-derived activated cells" (TDACs). Over 90% of the TDACs from each of the different tumor types were T lymphocytes, and the percentage of cells expressing either CD4 or CD8 varied considerably from population to population. The lymphocytes showed specific cytolytic activity in melanoma and colon and renal cell carcinomas. Continued expansion and long-term growth of the TDACs, as well as maintenance of the cytolytic activity, were achieved by periodic stimulation of the TDACs with irradiated autologous tumor cells. In a clinical study of 28 patients with cancer, we generated a mean number of 1.2 X 10(11) TDACs in an average time in culture of 69 days. These TDACs were subsequently infused into the patients with cancer. TDACs appear to represent an important resource for biotherapy of patients with cancer.

Cultured cell lines from human breast cancer biopsies and xenografts

“…Cell fines BRXBrl, BRXBr2, and BRXBr4 were used for the induction and screening of monoclonal antibodies for use in antibody-drug conjugate therapy. Cell lines BRXBr5, BRXBr6X, and BRXBr7 have been utilized in the induction and characterization of tumor-derived activated cells (TDAC) [4]. For this therapeutic approach, tumor mince was cultured in medium containing IL-2.…”

“…As the tumor cells in the lymphocyte cultures were killed, tumor cells were added back to stimulate the cell-killing specificity and expansion of the lymphocytes. Cultured tumor cells have served this purpose [4] for TDAC of breast cancer, and for several other types of cancer, such that cultured cells may replace the requirement for biopsy tumor cells in the development of TDAC cultures. As a result of successful tumor cell culture, the possibility of TDAC therapy can be extended to a greater number of cancer patients.…”

“…These therapies include active specific immunization [1], development of drugantibody conjugates [2], toxin-antibody conjugates [3], and tumor-derived activated cells [4]. Patients with advanced metastatic disease who have failed standard therapies are candidates for biological therapy.…”

Cultured breast cystosarcoma phylloides cells and applications to patient therapy

“…A preliminary assessment of growth response showed that two classes of mammary gland effector steroids, estrogen and progesterone, had no significant influence on growth of these cell lines in cul- [15,25] chromium release assay. 51Cr-labeled BT20 cells (2.5 × 103) were incubated with varying numbers of lymphocytes in the presence of I p~g/ml heteroconjugate antibody [26].…”

“…Tumor-derived activated lymphocyte cultures were prepared from previously cryopreserved tumor minces incubated with r-IL2 as described [15]. Briefly, lymphocytes (5 x 105 cells/ml) were grown in medium containing 10% human serum and 1000 U/ml r-IL2.…”