Activation and expansion of T cells

A method for expanding a population of T cells from a B-cell chronic lymphocytic leukemia (B-CLL) patient by cell surface moiety ligation, comprising:

• a. selecting a patient having B-CLL;
• b. providing a population of cells from the patient wherein at least a portion thereof comprises T cells;
• c. contacting said population of cells for about two hours with a surface, wherein said surface is a surface of a paramagnetic particle and wherein said surface has attached thereto a first agent that ligates a TCR/CD3 complex of a T cell in said population of cells, and a second agent that ligates a CD28 accessory molecule of said T cell;
• d. applying a magnetic force that predominantly drives simultaneous T cell concentration and ligation of the TCR/CD3 complex and the CD28 accessory molecule of said T cells;
• e. seeding said population of cells at a concentration of between about 0.2×106 and 5.0 ×106 cells/ml in a closed system comprising a disposable bioreactor bag comprising at least one inlet filter and one outlet filter and further contacting said T cells with said surface;
• f. perfusing medium through said closed system at about 1 ml/minute staffing at about day 8;
• g. rocking said bioreactor bag on a rocking platform at about 5-15 rocks/minute;
• and wherein said T cells expand to a concentration of about between 15 ×106 cells/ml to about 50×106 cells/ml in less than about two weeks.