In vitro assay measuring degree of activation of immune cells

A method for predicting clinical efficacy of immunotherapy of a patient with a sample of EVA immune cells in a primed state derived from said patient comprising:

• (a) contacting a portion of said sample of EVA immune cells with a single, intracellular-acting stimulant,
• wherein said stimulant at the dose used for stimulation will effectively stimulate primed immune cells to activity but wherein said stimulant does not effectively stimulate resting immune cells to activity;
• and,
• (b) measuring the activity of said EVA immune cells following stimulation;
• (c) contacting a sample of apheresis-derived peripheral blood mononuclear cells derived from said patient with said stimulant;
• (d) measuring the activity of said peripheral blood mononuclear cells;
• and
• (e) comparing the level of activity of the EVA immune cells with the level of activity of said peripheral blood mononuclear cells, wherein an increase in said level of activity of said EVA immune cells compared to said peripheral blood mononuclear cells indicates clinical efficacy of immunotherapy of said patient with said sample of EVA immune cells.

An assay to measure degree of activation of EVA immune cells in a primed state prior to reinfusion into a patient for adoptive immunotherapy comprising:

• (a) contacting said EVA immune cells with a single, intracellular-acting stimulant,
• wherein said stimulant at the dose used for stimulation will effectively stimulate primed immune cells to activity but wherein said stimulant does not effectively stimulate resting immune cells to activity;
• and,
• (b) measuring the degree of activation of the stimulated EVA immune cells;
• and
• (c) ascertaining whether the degree of activation of said stimulated EVA immune cells is predictive of in vivo efficacy for the patient prior to reinfusion of the EVA immune cells into the patient.

A method for measuring the ability of EVA immune cells in a primed state to enhance the activity of resting immune cells comprising:

• (a) culturing a mixture of resting immune cells and EVA immune cells;
• (b) stimulating the mixture to activity with a single, intracellular-acting stimulant;
• wherein said stimulant at the dose used for stimulation will effectively stimulate primed immune cells to activity but wherein said stimulant does not effectively stimulate resting immune cells to activity;
• and,
• (c) measuring the activity of said mixture following stimulation, wherein an increase in the activity of said mixture compared to the activity of resting immune cells in the absence of EVA cells indicates that said EVA cells enhance the activity of said resting immune cells.

An assay to measure degree of activation of EVA immune cells in a primed state prior to reinfusion into a patient for adoptive immunotherapy comprising:

• (a) stimulating EVA immune cells to activity with a single, intracellular-acting stimulant to produce stimulated EVA immune cells,
• wherein said stimulant at the dose used for stimulation will effectively stimulate the primed immune cells to activity but wherein said stimulant does not effectively stimulate resting immune cells to activity;
• and,
• (b) measuring the degree of activation of the stimulated EVA immune cells, and
• (c) comparing the degree of activation of said stimulated EVA immune cells to a minimum standard to predict whether infusion of the EVA cells into the patient will be efficacious.

A method of assessing the immune cell status of a patient, wherein the immune cells are obtained from the patient comprising:

• (a) obtaining a sample of immune cells from the patient;
• (b) stimulating the immune cells to activity with a single, intracellular-acting stimulant;
• wherein said stimulant at the dose used for stimulation will effectively stimulate primed immune cells to activity but wherein said stimulant does not effectively stimulate resting immune cells to activity;
• (c) measuring the activity of said immune cells following stimulation;
• and
• (d) comparing said activity to a minimum standard as an indication of the immune status of a patient undergoing treatment with EVA cells.