Rapid expansion method ("REM") for in vitro propagation of T lym

Details Rapid expansion method ("REM") for in vitro propagation of T lym Rapid expansion method ("REM") for in vitro propagation of T lym

1994-10-03

1998-10-27

Schwadron, Ronald B.

Chemistry: molecular biology and microbiology

Maintaining blood or sperm in a physiologically active state...

4353723, 435373, 435375, 435383, 435386, 435384, 424 9371, C12N 502, C12N 508, A01N 102, A61K 3512

Patent

active

058276424

ABSTRACT:
The present invention provides a rapid expansion method (termed "REM"), for quickly generating large numbers of T lymphocytes, including cytolytic and helper T lymphocytes. REM involves culturing the T cells in association with a disproportionately large concentration of nondividing feeder cells, preferably .gamma.-irradiated peripheral blood mononuclear cells ("PBMC") present at an excess of at least 40-fold (relative to the number of target T cells), more preferably at an excess of at least about 200-fold. Cultures grown under REM exhibit dramatically enhanced expansion rates that can be even further elevated by the use of appropriate concentrations of an additional feeder cell, an anti-CD3 monoclonal antibody and IL-2, as described herein. Clonal expansions in the range of 500-fold to 3000-fold can be achieved within a single stimulation cycle of about 10-13 days, which is more than 100-fold more efficient than currently employed methods of culturing human T cell clones. Genetic transduction efficiencies were also enhanced using REM-expanded T lymphocytes. Several examples involving human bone marrow transplant recipients illustrate the effective use of REM-expanded antigen-specific cytotoxic T lymphocytes for adoptive immunotherapy in humans.

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