Method for ex vivo immunization using heterologous intact bispecific and/or trispecific antibodies

A composition comprising activated peripheral blood mononucleated cells and non-viable tumor cells from the same individual, the composition obtained by incubating ex vivo:

• a) tumor cells isolated from a patient and treated to prevent survival following reinfusion into the patient;
• b) peripheral blood mononucleated cells from the patient;
• and
• c) an intact heterologous bispecific antibody showing the following properties:
• (i) binding to a T cell;
• (ii) binding to at least one tumor-associated antigen on the tumor cells;
• (iii) binding, by their Fc portion to Fc receptor-positive cells;
• and
• (iv) activating the Fc receptor-positive cells, thereby inducing or increasing the expression of a cytokine and/or a co-stimulatory antigen,
• wherein the bispecific antibody has an isotype combination selected from the group consisting of:
• rat-IgG2b/human-IgG1,
• rat-IgG2b/human-IgG2,
• rat-IgG2b/human-IgG3[oriental allotype G3m(st)=binding to protein A],
• rat-IgG2b/human-IgG4,
• rat-IgG2b/rat-IgG2c,
• mouse-IgG2a/human-IgG3[caucasian allotypes G3m(b+g)=no binding to protein A, in the following indicated as *],
• mouse-IgG2a/mouse-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-IgG2a/rat-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-IgG2a/human-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-[VH-CH1,VL-CL]-human-IgG1/rat-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-[VH-CH1,VL-CL]-human-IgG4/rat-[VH-CH1,VL-CL]-human-IgG4-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2:
• >aa position 251]-human-IgG3*[CH3],
• rat-IgG2b/mouse-[VH-CH1,VL-CL]-human-IgG1-[hinge-CH2-CH3],
• rat-IgG2b/mouse-[VH-CH1,VL-CL]-human-IgG2-[hinge-CH2-CH3],
• rat-IgG2b/mouse-[VH-CH1,VL-CL]-human-IgG3-[hinge-CH2-CH3, oriental allotype],
• rat-IgG2b/mouse-[VH-CH1,VL-CL]-human-IgG4-[hinge-CH2-CH3],
• human-IgG1/human-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• human-IgG1/rat-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2:
• >aa position 251]-human-IgG3*[CH3],
• human-IgG1/mouse-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2:
• >aa position 251]-human-IgG3*[CH3],
• human-IgG1/rat-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG2[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2:
• >aa position 251]-human-IgG3*[CH3],
• human-IgG1/mouse-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG2[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2:
• >aa position 251]-human-IgG3*[CH3],
• human-IgG1/rat-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• human-IgG1/mouse-[VH-CH1, VL-CL]-human-IgG-[hinge]-human-IgG3*-[CH2-CH3],
• human-IgG2/human-[VH-CH1,VL-CL]-human-IgG2-[hinge]-human-IgG3*-[CH2-CH3],
• human-IgG4/human-[VH-CH1,VL-CL]-human-IgG4-[hinge]-human-IgG3*-[CH2-CH3],
• human-IgG4/human-[VH-CH1,VL-CL]-human-IgG4-[hinge]-human-IgG4[N-terminal region of CH2]-human-IgG3*[C-terminal region of CH2:
• >aa position 251]-human-IgG3*[CH3],
• mouse-IgG2b/rat-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-IgG2b/human-[VH-CH1, VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-IgG2b/mouse-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG3*-[CH2-CH3],
• mouse-[VH-CH1,VL-CL]-human-IgG4/rat-[VH-CH1,VL-CL]-human-IgG4-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3],
• human-IgG1/rat-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3],
• human-IgG1/mouse-[VH-CH1,VL-CL]-human-IgG1-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3],
• human-IgG4/human-[VH-CH1,VL-CL]-human-IgG4-[hinge]-human-IgG4-[CH2]-human-IgG3*-[CH3],
• rat-IgG2b/mouse-IgG2a,
• rat-IgG2b/mouse-IgG2b, and
• rat-IgG2b/mouse-IgG3.