Scaffolds to treat solid tumor cells and escape variants

Abstract

Implantable scaffolds that treat solid tumors and escape variants and that provide effective vaccinations against cancer recurrence are described. The scaffolds include genetically-reprogrammed lymphocytes and a lymphocyte-activating moiety.

1 . A lymphocyte scaffold consisting of a scaffold matrix, genetically reprogrammed lymphocytes, and three lymphocyte-activating moieties. 
     
     
         2 . The lymphocyte scaffold of  claim 1 , wherein the scaffold matrix is an alginate scaffold, a collagen/alginate scaffold, a chitosan scaffold, a self-assembling peptide scaffold, a mesoporous silica scaffold, or a PLGA scaffold. 
     
     
         3 . The lymphocyte scaffold of  claim 1 , wherein the scaffold matrix is an alginate scaffold. 
     
     
         4 . The lymphocyte scaffold of  claim 1 , wherein the scaffold matrix is a polymeric calcium cross-linked alginate scaffold. 
     
     
         5 . The lymphocyte scaffold of  claim 1 , wherein the genetically reprogrammed lymphocytes are genetically reprogrammed T-cells and/or natural killer cells. 
     
     
         6 . The lymphocyte scaffold of  claim 1 , wherein the genetically reprogrammed lymphocytes are CD8+ T-cells. 
     
     
         7 . The lymphocyte scaffold of  claim 1 , comprising at least 2×10 6  genetically reprogrammed lymphocytes. 
     
     
         8 . The lymphocyte scaffold of  claim 1 , comprising at least 7×10 6  genetically reprogrammed lymphocytes. 
     
     
         9 . The lymphocyte scaffold of  claim 1 , wherein the lymphocyte-activating moieties comprise antibodies specific for CD3, CD28, and CD137. 
     
     
         10 . The lymphocyte scaffold of  claim 1 , wherein the lymphocyte-activating moieties are linked to the scaffold. 
     
     
         11 . The lymphocyte scaffold of  claim 1 , wherein the lymphocyte-activating moieties are embedded within the scaffold. 
     
     
         12 . A lymphocyte scaffold comprising: a scaffold matrix, genetically-reprogrammed lymphocytes, and a lymphocyte-activating moiety. 
     
     
         13 . The lymphocyte scaffold of  claim 12 , further comprising a STING agonist. 
     
     
         14 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist comprises c-diGMP, c-diAMP, c-GAMP, c-AIMP, (3′,2′)c-AIMP, (2′,2′)c-AIMP, (2′,3′)c-AIMP, c-AIMP(S), c-(dAMP-dIMP), c-(dAMP-2′FdIMP), c-(2′FdAMP-2′FdIMP), (2′,3′)c-(AMP-2′FdIMP), c-[2′FdAMP(S)-2′FdIMP(S)], c-[2′FdAMP(S)-2′FdIMP(S)](POM)2, and/or DMXAA. 
     
     
         15 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist comprises c-diGMP. 
     
     
         16 . The lymphocyte scaffold of  claim 12 , wherein the genetically-reprogrammed lymphocytes are genetically-reprogrammed T-cells and/or natural killer cells. 
     
     
         17 . The lymphocyte scaffold of  claim 12 , wherein the genetically-reprogrammed lymphocytes are CD8+ T-cells. 
     
     
         18 . The lymphocyte scaffold of  claim 12 , comprising at least 7×10 6  genetically-reprogrammed lymphocytes. 
     
     
         19 . The lymphocyte scaffold of  claim 12 , wherein the lymphocyte-activating moiety comprises antibodies specific for CD3, CD28, and/or CD137. 
     
     
         20 . The lymphocyte scaffold of  claim 12 , further comprising an immune stimulant. 
     
     
         21 . The lymphocyte scaffold of  claim 20 , wherein the immune stimulant is a cytokine, an antibody, a small molecule, an siRNA, a plasmid DNA, and/or a vaccine adjuvant. 
     
     
         22 . The lymphocyte scaffold of  claim 20 , wherein the immune stimulant is selected from (i) a Toll-like receptor ligand selected from CpG, Cpg-28, Poly(I:C), α-galactoceramide, MPLA, VTX-2337, EMD1201081) imiquimod, MGN1703, G100, CBLB502, Hiltonol, and Imiquimod, and/or (ii) 17-dimethylaminoethylamino-17-demethoxygeldanamycin). 
     
     
         23 . The lymphocyte scaffold of  claim 12 , further comprising a lymphocyte-adhesion moiety. 
     
     
         24 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises fibrin. 
     
     
         25 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises a peptide that binds αιβι integrin, α2βι integrin, α4βι integrin, α5βι integrin, or lymphocyte function associated antigen (LFA-1). 
     
     
         26 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises a GFOGER (SEQ ID NO: 1) peptide. 
     
     
         27 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises a GFOGER (SEQ ID NO: 1) peptide of SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         28 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises an ICAM-1 peptide. 
     
     
         29 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises an ICAM-1 peptide of SEQ ID NO: 3. 
     
     
         30 . The lymphocyte scaffold of any  claim 23 , wherein the lymphocyte-adhesion moiety comprises a FNIII 7 10 peptide. 
     
     
         31 . The lymphocyte scaffold of  claim 23 , wherein the lymphocyte-adhesion moiety comprises a FNIII 7-10  peptide of SEQ ID NO: 4. 
     
     
         32 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist and/or the lymphocyte-activating moiety is linked to the scaffold matrix. 
     
     
         33 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist and/or the lymphocyte-activating moiety is embedded within the scaffold matrix. 
     
     
         34 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist and/or the lymphocyte-activating moiety is within a bioactive coating overlaying at least a portion of the surface of the scaffold matrix. 
     
     
         35 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist and/or the lymphocyte-activating moiety is linked to particles. 
     
     
         36 . The lymphocyte scaffold of  claim 35 , wherein the particles are linked to the scaffold matrix and/or embedded within the scaffold matrix. 
     
     
         37 . The lymphocyte scaffold of  claim 13 , wherein the STING agonist and/or the lymphocyte-activating moiety is bound to a liposome of a protocell. 
     
     
         38 . The lymphocyte scaffold of  claim 37 , wherein the ratio of the protocells to the lymphocytes within the scaffold matrix is 0.5:1; 1:1; 5:1; or 10:1. 
     
     
         39 . The lymphocyte scaffold of  claim 12 , wherein the lymphocyte scaffold comprises 7×10 6  to 1×10 10  protocells. 
     
     
         40 . The lymphocyte scaffold of  claim 12 , wherein the scaffold matrix is an alginate scaffold, a collagen/alginate scaffold, a chitosan scaffold, a self-assembling peptide scaffold, a mesoporous silica scaffold, or a PLGA scaffold. 
     
     
         41 . The lymphocyte scaffold of  claim 12 , wherein the scaffold matrix is an alginate scaffold. 
     
     
         42 . The lymphocyte scaffold of  claim 12 , wherein the scaffold matrix is a polymeric calcium cross-linked alginate scaffold. 
     
     
         43 . A lymphocyte scaffold comprising:
 (i) a scaffold matrix comprising an alginate scaffold;   (ii) genetically-reprogrammed lymphocytes;   (iii) a lymphocyte-adhesion moiety comprising a GFOGER (SEQ ID NO: 1) peptide;   (iv) lymphocyte-activating moieties comprising antibodies specific for CD3, CD28, and CD137; and   (v) a STING agonist.   
     
     
         44 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist comprises c-diGMP, c-diAMP, c-GAMP, c-AIMP, (3′,2′)c-AIMP, (2′,2′)c-AIMP, (2′,3′)c-AIMP, c-AIMP(S), c-(dAMP-dIMP), c-(dAMP-2′FdIMP), c-(2′FdAMP-2′FdIMP), (2′,3′)c-(AMP-2′FdIMP), c-[2′FdAMP(S)-2′FdIMP(S)], c-[2′FdAMP(S)-2′FdIMP(S)](POM)2, and/or DMXAA. 
     
     
         45 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist comprises c-diGMP. 
     
     
         46 . The lymphocyte scaffold of  claim 43 , wherein the genetically-reprogrammed lymphocytes are genetically-reprogrammed T-cells and/or natural killer cells. 
     
     
         47 . The lymphocyte scaffold of  claim 43 , wherein the genetically-reprogrammed lymphocytes are CD8+ T-cells. 
     
     
         48 . The lymphocyte scaffold of  claim 43 , comprising at least 7×10 6  genetically-reprogrammed lymphocytes. 
     
     
         49 . The lymphocyte scaffold of  claim 43 , further comprising an immune stimulant. 
     
     
         50 . The lymphocyte scaffold of  claim 49 , wherein the immune stimulant is a cytokine, an antibody, a small molecule, an siRNA, a plasmid DNA, and/or a vaccine adjuvant. 
     
     
         51 . The lymphocyte scaffold of  claim 49 , wherein the immune stimulant is selected from (i) a Toll-like receptor ligand selected from CpG, Cpg-28, Poly(I:C), α-galactoceramide, MPLA, VTX-2337, EMD1201081) imiquimod, MGN1703, G100, CBLB502, Hiltonol, and Imiquimod, and/or (ii) 17-dimethylaminoethylamino-17-demethoxygeldanamycin). 
     
     
         52 . The lymphocyte scaffold of  claim 43 , wherein the GFOGER (SEQ ID NO: 1) peptide comprises a peptide of SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         53 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist and/or the lymphocyte-activating moieties are linked to the scaffold matrix. 
     
     
         54 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist and/or the lymphocyte-activating moieties are embedded within the scaffold matrix. 
     
     
         55 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist and/or the lymphocyte-activating moieties are within a bioactive coating overlaying at least a portion of the surface of the scaffold matrix. 
     
     
         56 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist and/or the lymphocyte-activating moieties are linked to particles. 
     
     
         57 . The lymphocyte scaffold of  claim 56 , wherein the particles are linked to the scaffold matrix and/or embedded within the scaffold matrix. 
     
     
         58 . The lymphocyte scaffold of  claim 43 , wherein the STING agonist and/or the lymphocyte-activating moieties are bound to a liposome of a protocell. 
     
     
         59 . The lymphocyte scaffold of  claim 58 , wherein the ratio of the protocells to the lymphocytes within the scaffold matrix is 0.5:1; 1:1; 5:1; or 10:1. 
     
     
         60 . The lymphocyte scaffold of  claim 58 , wherein the lymphocyte scaffold comprises 7×10 6  to 1×10 10  protocells. 
     
     
         61 . The lymphocyte scaffold of  claim 43 , wherein the alginate scaffold is a polymeric calcium cross-linked alginate scaffold. 
     
     
         62 . A lymphocyte scaffold comprising: (i) a scaffold matrix material, (ii) natural killer cells with anti-cancer activity, and (iii) lymphocyte-activating moieties comprising IL-15 and an antibody specific to CD137. 
     
     
         63 . The lymphocyte scaffold of  claim 62 , further comprising a STING agonist. 
     
     
         64 . The lymphocyte scaffold of  claim 63 , wherein the STING agonist comprises c-diGMP, c-diAMP, c-GAMP, c-AIMP, (3′,2′)c-AIMP, (2′,2′)c-AIMP, (2′,3′)c-AIMP, c-AIMP(S), c-(dAMP-dIMP), c-(dAMP-2′FdIMP), c-(2′FdAMP-2′FdIMP), (2′,3′)c-(AMP-2′FdIMP), c-[2′FdAMP(S)-2′FdIMP(S)], c-[2′FdAMP(S)-2′FdIMP(S)](POM)2, and/or DMXAA. 
     
     
         65 . The lymphocyte scaffold of  claim 62 , further comprising a drug-eluting polymer. 
     
     
         66 . The lymphocyte scaffold of  claim 65 , wherein the drug eluting polymer comprises PLGA. 
     
     
         67 . The lymphocyte scaffold of  claim 62 , wherein the STING agonist is embedded within a drug eluting polymer. 
     
     
         68 . The lymphocyte scaffold of  claim 62 , further comprising a lymphocyte-adhesion moiety. 
     
     
         69 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises fibrin. 
     
     
         70 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises a peptide that binds αιβι integrin, α2βι integrin, α4βι integrin, α5βι integrin, or lymphocyte function associated antigen (LFA-1). 
     
     
         71 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises a GFOGER (SEQ ID NO: 1) peptide. 
     
     
         72 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises a GFOGER (SEQ ID NO: 1) peptide of SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         73 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises an ICAM-1 peptide. 
     
     
         74 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises an ICAM-1 peptide of SEQ ID NO: 3. 
     
     
         75 . The lymphocyte scaffold of any  claim 68 , wherein the lymphocyte-adhesion moiety comprises a FNIII 7 10 peptide. 
     
     
         76 . The lymphocyte scaffold of  claim 68 , wherein the lymphocyte-adhesion moiety comprises a FNIII 7-10  peptide of SEQ ID NO: 4. 
     
     
         77 . A method of treating a heterogenous solid tumor comprising escape variant tumor cells in a subject comprising implanting a lymphocyte scaffold of  claim 62  into the subject within a proximity of the heterogenous solid tumor cell sufficient to lead to the destruction of the heterogenous solid tumor in the subject, thereby treating the heterogenous solid tumor comprising escape variant tumor cells. 
     
     
         78 . A method of vaccinating a subject against development of cancer recurrence comprising implanting a lymphocyte scaffold of  claim 62  into the subject within a proximity of a heterogenous solid tumor or within a solid tumor resection bed in the subject, thereby vaccinating the subject against development of cancer recurrence. 
     
     
         79 . A method of treating tumor cells in a subject in need thereof comprising implanting a lymphocyte scaffold of  claim 62  into the subject within a tumor resection bed thereby treating the tumor cells in the subject. 
     
     
         80 . The method of  claim 79 , wherein a treated tumor cell is an adrenal cancer cell, a brain cancer cell, a breast cancer cell, a cervical cancer cell, a colon cancer cell, a colorectal cancer cell, an ear, nose and throat (ENT) cancer cell, an endometrial cancer cell, an esophageal cancer cell, a gastrointestinal cancer cell, a glioma cell, a head and neck cancer cell, an intestinal cancer cell, a kidney cancer cell, a liver cancer cell, a lung cancer cell, a lymph node cancer cell, a melanoma cell, a neuroblastoma cell, an ovarian cancer cell, a pancreatic cancer cell, a prostate cancer cell, a rectal cancer cell, a seminoma cell, a skin cancer cell, a stomach cancer cell, a teratoma cell, a thyroid cancer cell, or a uterine cancer cell. 
     
     
         81 . The method of  claim 79 , wherein a treated tumor cell is a glioblastoma cell, a pancreatic adenocarcinoma cell, or an ovarian cancer cell. 
     
     
         82 . A method of reducing surgical treatment failure caused by metastatic relapse after resection of a primary tumor, comprising administering a lymphocyte scaffold of  claim 62  to a tumor resection bed of a subject thereby reducing surgical treatment failure caused by metastatic relapse after primary tumor resection. 
     
     
         83 . The method of  claim 82 , wherein the primary tumor comprises a seminoma cell, a melanoma cell, a teratoma cell, a neuroblastoma cell, a glioma cell, a rectal cancer cell, an endometrial cancer cell, a kidney cancer cell, an adrenal cancer cell, a thyroid cancer cell, a skin cancer cell, a brain cancer cell, a cervical cancer cell, an intestinal cancer cell, a liver cancer cell, a colon cancer cell, a stomach cancer cell, a head and neck cancer cell, a gastrointestinal cancer cell, a lymph node cancer cell, an esophageal cancer cell, a colorectal cancer cell, a pancreatic cancer cell, an ear, nose and throat (ENT) cancer cell, a breast cancer cell, a prostate cancer cell, a uterine cancer cell, an ovarian cancer cell, or a lung cancer cell. 
     
     
         84 . A kit to form a lymphocyte scaffold to treat a solid tumor in a subject comprising (i) a scaffold matrix; and (ii) lymphocyte-activating moieties comprising antibodies specific for CD3, CD28, and CD137. 
     
     
         85 . The kit of  claim 84 , further comprising porous particles. 
     
     
         86 . The kit of  claim 84 , further comprising liposomes. 
     
     
         87 . The kit of  claim 84 , further comprising protocells. 
     
     
         88 . The kit of  claim 84 , further comprising a STING agonist. 
     
     
         89 . The kit of  claim 88 , wherein the STING agonist comprises c-diGMP, c-diAMP, c-GAMP, c-AIMP, (3′,2′)c-AIMP, (2′,2′)c-AIMP, (2′,3′)c-AIMP, c-AIMP(S), c-(dAMP-dIMP), c-(dAMP-2′FdIMP), c-(2′FdAMP-2′FdIMP), (2′,3′)c-(AMP-2′FdIMP), c-[2′FdAMP(S)-2′FdIMP(S)], c-[2′FdAMP(S)-2′FdIMP(S)](POM)2, and/or DMXAA. 
     
     
         90 . The kit of  claim 88 , wherein the STING agonist comprises c-diGMP. 
     
     
         91 . The kit of  claim 84 , further comprising an immune stimulant. 
     
     
         92 . The kit of  claim 91 , wherein the immune stimulant is a cytokine, an antibody, a small molecule, an siRNA, a plasmid DNA, and/or a vaccine adjuvant. 
     
     
         93 . The kit of  claim 91 , wherein the immune stimulant is selected from (i) a Toll-like receptor ligand selected from CpG, Cpg-28, Poly(I:C), α-galactoceramide, MPLA, VTX-2337, EMD1201081) imiquimod, MGN1703, G100, CBLB502, Hiltonol, and Imiquimod, and/or (ii) 17-dimethylaminoethylamino-17-demethoxygeldanamycin). 
     
     
         94 . The kit of  claim 84 , further comprising a lymphocyte-adhesion moiety. 
     
     
         95 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises fibrin. 
     
     
         96 . The kit of  claim 4 , wherein the lymphocyte-adhesion moiety comprises a peptide that binds αιβι integrin, α2βι integrin, α4βι integrin, α5βι integrin, or lymphocyte function associated antigen (LFA-1). 
     
     
         97 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises a GFOGER (SEQ ID NO: 1) peptide. 
     
     
         98 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises a GFOGER (SEQ ID NO: 1) peptide of SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         99 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises an ICAM-1 peptide. 
     
     
         100 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises an ICAM-1 peptide of SEQ ID NO: 3. 
     
     
         101 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises a FNIII 7-10  peptide. 
     
     
         102 . The kit of  claim 94 , wherein the lymphocyte-adhesion moiety comprises a FNIII 7 10 peptide of SEQ ID NO: 4. 
     
     
         103 . The kit of  claim 88 , wherein the kit comprises 7×10 6  to 1×10 10  particles, liposomes or protocells. 
     
     
         104 . The kit of  claim 88 , further comprising genetically-reprogrammed lymphocytes. 
     
     
         105 . The kit of  claim 104 , wherein the genetically-reprogrammed lymphocytes are T-cells and/or natural killer cells. 
     
     
         106 . The kit of  claim 104 , wherein the genetically-reprogrammed lymphocytes are CD8+ T-cells. 
     
     
         107 . The kit of  claim 104 , wherein the lymphocytes comprise at least 7×10 6  lymphocytes. 
     
     
         108 . The kit of  claim 88 , wherein the scaffold matrix comprises alginate, collagen, chitosan, a self-assembling peptide, mesoporous silica, or PLGA. 
     
     
         109 . The kit of  claim 108 , wherein the scaffold matrix comprises alginate. 
     
     
         110 . The kit of  claim 88 , further comprising calcium. 
     
     
         111 . The kit of  claim 88 , further comprising a drug eluting polymer. 
     
     
         112 . The kit of  claim 111 , wherein the drug eluting polymer is coated on the scaffold matrix. 
     
     
         113 . The kit of  claim 111 , wherein a STING agonist or an immune stimulant is embedded within the drug eluting polymer. 
     
     
         114 . The kit of  claim 111 , wherein the drug eluting polymer comprises PLGA. 
     
     
         115 . A kit to form a lymphocyte scaffold comprising: (i) a scaffold matrix comprising an alginate scaffold, (ii) a lymphocyte-adhesion moiety comprising a GFOGER (SEQ ID NO: 1) peptide, (iii) lymphocyte-activating moieties comprising antibodies specific for CD3, CD28, and CD137, and (iv) a STING agonist. 
     
     
         116 . The kit of  claim 115 , further comprising porous particles. 
     
     
         117 . The kit of  claim 115 , further comprising liposomes. 
     
     
         118 . The kit of  claim 115 , further comprising protocells. 
     
     
         119 . The kit of claim  1150 , wherein the STING agonist comprises c-diGMP, c-diAMP, c-GAMP, c-AIMP, (3′,2′)c-AIMP, (2′,2′)c-AIMP, (2′,3′)c-AIMP, c-AIMP(S), c-(dAMP-dIMP), c-(dAMP-2′FdIMP), c-(2′FdAMP-2′FdIMP), (2′,3′)c-(AMP-2′FdIMP), c-[2′FdAMP(S)-2′FdIMP(S)], c-[2′FdAMP(S)-2′FdIMP(S)](POM)2, and/or DMXAA. 
     
     
         120 . The kit of  claim 115 , wherein the STING agonist comprises c-diGMP. 
     
     
         121 . The kit of  claim 115 , further comprising an immune stimulant. 
     
     
         122 . The kit of  claim 121 , wherein the immune stimulant is a cytokine, an antibody, a small molecule, an siRNA, a plasmid DNA, and/or a vaccine adjuvant. 
     
     
         123 . The kit of  claim 121 , wherein the immune stimulant is selected from (i) a Toll-like receptor ligand selected from CpG, Cpg-28, Poly(I:C), α-galactoceramide, MPLA, VTX-2337, EMD1201081) imiquimod, MGN1703, G100, CBLB502, Hiltonol, and Imiquimod, and/or (ii) 17-dimethylaminoethylamino-17-demethoxygeldanamycin). 
     
     
         124 . The kit of  claim 115 , wherein the kit comprises 7×10 6  to 1×10 10  particles, liposomes or protocells. 
     
     
         125 . The kit of  claim 115 , further comprising a vector for genetically reprogrammed lymphocytes. 
     
     
         126 . The kit of  claim 115 , further comprising genetically-reprogrammed lymphocytes. 
     
     
         127 . The kit of  claim 126 , wherein the genetically-reprogrammed lymphocytes are T-cells and/or natural killer cells. 
     
     
         128 . The kit of  claim 126 , wherein the genetically-reprogrammed lymphocytes are CD8+ T-cells. 
     
     
         129 . The kit of  claim 126 , wherein the lymphocytes comprise at least 7×10 6  lymphocytes. 
     
     
         130 . The kit of  claim 115 , further comprising calcium.