PURPOSE:

Engineering T cells with receptors to re-direct the immune system against cancer has most recently been described as a scientific breakthrough. However, a main challenge remains the GMP-grade purification of immune cells selectively expressing the introduced receptor in order to reduce potential side effects due to poorly or non-engineered cells.

EXPERIMENTAL DESIGN:

In order to test a novel purification strategy, we took advantage of a model γδT cell receptor (TCR), naturally interfering with endogenous αβTCR expression and designed the optimal retroviral expression cassette to achieve maximal interference with endogenous αβTCR chains. Following retroviral transduction, non-and poorly engineered immune cells characterized by a high endogenous αβTCR expression were efficiently depleted with GMP-grade anti-αβTCR-beads. Next, the engineered immune cells were validated for TCR expression, function against a panel of tumor cell lines and primary tumors and potential allo-reactivity. Engineered immune cells were further validated in two humanized mouse tumor models.

 

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