Use of Triplex CMV Vaccine in CAR T Cell Therapy

1 - 50 . (canceled)

51 . A method for treating a patient comprising:

(a) providing a composition comprising a population of T cells expressing both a chimeric antigen receptor (CAR) and a T cell receptor specific for a cytomegalovirus (CMV) antigen;

(b) administering the composition of part (a) to the patient; and

(c) administering to the patient a viral vector encoding:

(i) CMV pp65 and

(ii) a fusion protein comprising exon 4 of CMV protein IE1 (e4) and exon 5 of CMV protein IE2 (e5).

52 . The method of claim 51 , wherein the viral vector of part (a) is a MVA virus.

53 . The method of claim 52 , wherein expression of (i) CMV pp65 and (ii) the fusion protein comprising exon 4 of CMV protein IE1 (e4) and exon 5 of CMV protein IE2 (e5) is under the control of mH5 promoter.

54 . The method of claim 51 , wherein the patient is CMV-seronegative prior to treatment.

55 . The method of claim 51 , wherein the patient is CMV-seropositive prior to treatment.

56 . The method of claim 51 , wherein the CAR is targeted to CD19.

57 . The method of claim 51 , wherein the viral vector is administered to the patient both prior to and subsequent to the administration of the composition comprising a population of T cells.

58 . The method of claim 51 , wherein the step of providing a population of T cells expressing a CAR and a T cell receptor specific for a CMV antigen comprises:

(a1) providing PBMC or a T cell subpopulation from a CMV-seropositive human donor;

(a2) exposing the PBMC or the T cell subpopulation of part (a1) to at least one CMV antigen;

(a3) treating the exposed cells of part (a2) to produce a population of cells enriched for cells specific for CMV; and

(a4) transducing at least a portion of the enriched population of cells of part (a3) with a vector expressing a CAR.

59 . The method of claim 58 , wherein the step of treating the exposed cells to produce a population of cells enriched for cells specific for CMV comprises treating the cells to produce a population of cells enriched for cells expressing an activation marker.

60 . The method of claim 51 , wherein the step of providing a population of T cell expressing a CAR and a T cell receptor specific for a CMV antigen further comprises:

(a1) administering a viral vector encoding: (i) CMV pp65 and (ii) a fusion protein comprising exon 4 of CMV protein IE1 (e4) and exon 5 of CMV protein IE2 (e5) to a human donor to convert a CMV-seronegative human donor to one containing T cells responsive to CMV antigens pp65, IE1 and IE2;

(a2) obtaining PBMC from the CMV-seropositive human donor of (a1);

(a3) exposing the PBMC of (a2) to at least one CMV antigen;

(a4) treating the exposed cells of (a3) to produce a population of cells enriched for stimulated cells specific for CMV; and

(a5) transducing at least a portion of the enriched population of cells of (a4) with a vector expressing a CAR, thereby providing a population of T cell expressing a CAR and a T cell receptor specific for a CMV antigen.

61 . The method of claim 51 , wherein the step of providing a population of T cell expressing a CAR and a T cell receptor specific for a CMV antigen comprises:

(a1) administering a viral vector encoding: (i) CMV pp65 and (ii) a fusion protein comprising exon 4 of CMV protein IE1 (e4) and exon 5 of CMV protein IE2 (e5) to a CMV-positive human donor;

(a2) obtaining PBMC from the CMV-seropositive human donor of (a1);

(a3) exposing the PBMC of (a2) to at least one CMV antigen;

(a4) treating the exposed cells of (a3) to produce a population of cells enriched for stimulated cells specific for CMV;

(a5) transducing at least a portion of the enriched population of cells of (a4) with a vector expressing a CAR, thereby providing a population of T cell expressing a CAR and a T cell receptor specific for a CMV antigen.

62 . A method for preparing T cells expressing a CAR and a T cell receptor specific for a CMV antigen, the method comprising:

(a) administering a viral vector encoding: (i) CMV pp65 and (ii) a fusion protein comprising exon 4 of CMV protein IE1 (e4) and exon 5 of CMV protein IE2 (e5) to a CMV-positive human donor;

(b) obtaining PBMC from the CMV-seropositive human donor;

(c) exposing the PBMC to at least one CMV antigen;

(d) treating the exposed cells to produce a population of cells enriched for stimulated cells specific for CMV; and

(e) transducing at least a portion of the enriched population of cells with a vector expressing a CAR, thereby providing a population of T cell expressing a CAR and a T cell receptor specific for a CMV antigen.

63 . A method for preparing T cells expressing a CAR and a T cell receptor specific for a CMV antigen, the method comprising:

(a) administering a viral vector encoding: (i) CMV pp65 and (ii) a fusion protein comprising exon 4 of CMV protein IE1 (e4) and exon 5 of CMV protein IE2 (e5) to a CMV-positive human donor;

(b) obtaining PBMC from the CMV-seropositive human donor;

(c) (b) exposing the PBMC to at least one CMV antigen;

(d) treating the exposed cells to produce a population of cells enriched for stimulated cells specific for CMV; and

(e) transducing at least a portion of the enriched population of cells with a vector expressing a CAR, thereby providing a population of T cell expressing a CAR and a T cell receptor specific for a CMV antigen.

64 . The method of claim 51 , wherein the CAR is selective for an antigen selected from: CD19, CD123, CS1, BCMA, CD44v6, CD33, CD22, IL-13α2, PSA, HER-2, EGFRv3, CEA, and C7R.

65 . The method of claim 51 , wherein the CAR comprises: a scFv selective for a non-CMV antigen; a hinge/linker region; a transmembrane domain; a co-signaling domain; and

CD3 ζ signaling domain.

66 . The method of claim 65 , wherein the co-signaling domain is selected from a CD28 co-signaling domain and a 4-IBB co-signaling domain.

67 . The method of claim 65 , wherein the transmembrane domain is selected from a CD28 transmembrane domain and a CD4 transmembrane