Lymphangiogenesis for therapeutic immunomodulation

Abstract

The present invention concerns methods and compositions for evoking protective immune responses against pathogen infection or cancer. In certain embodiments, the methods and compositions comprise a lymphangiogenesis inducer and an antigen.

1 . A method of eliciting an immune response to an antigen in a subject comprising administering to the subject one or more lymphangiogenesis inducers and an effective amount of the antigen. 
     
     
         2 . The method of  claim 1 , wherein the one or more lymphangiogenesis inducers comprise vascular endothelial growth factor C (VEGF-C) or vascular endothelial growth factor D (VEGF-D). 
     
     
         3 . The method of  claim 1  or  2 , wherein the one or more lymphangiogenesis inducers comprise CCL21. 
     
     
         4 . The method of any of  claims 1 - 3 , wherein the antigen is bacterial antigen, a viral antigen, a fungal antigen, a protozoal antigen, a helminth antigen or a cancer antigen. 
     
     
         5 . The method of any of  claims 1 - 4 , wherein the lymphangiogenesis inducer and the antigen are administered in a single composition. 
     
     
         6 . The method of any of  claims 1 - 5 , wherein the lymphangiogenesis inducer and the antigen are provided over multiple administrations. 
     
     
         7 . The method of any of  claims 1 - 6 , wherein the subject is also administered an adjuvant. 
     
     
         8 . The method of any of  claims 1 - 7 , wherein the lymphangiogenesis inducer and the antigen are administered in a single composition comprising an adjuvant. 
     
     
         9 . The method of any of  claims 1 - 8 , wherein the lymphangiogenesis inducer and the antigen are administered in a single composition comprising a pharmaceutically acceptable excipient. 
     
     
         10 . The method of any of  claims 5 - 9 , wherein the composition is administered parenterally, subcutaneously or intramuscularly. 
     
     
         11 . The method of any of  claims 1 - 10 , wherein the subject is administered an effective amount of a second antigen. 
     
     
         12 . The method of any of  claims 1 - 11 , wherein the lymphangiogenesis inducer is incorporated into a matrix. 
     
     
         13 . The method of  claim 12 , wherein the antigen is bound or incorporated into the matrix and is cleavable. 
     
     
         14 . The method of  claims 12 - 13 , wherein the matrix incorporates one or more cleavable chemoattractants or one or more cleavable cytokines. 
     
     
         15 . The method of any one of  claims 12 - 14 , wherein the matrix is a hydrogel. 
     
     
         16 . The method of  claim 15 , wherein the hydrogel is a fibrin hydrogel or a fibrin domain modified polyethylene glycol hydrogel. 
     
     
         17 . The method of any of  claims 12 - 16 , wherein the lymphangiogenesis inducer is capable of binding to the matrix or hydrogel. 
     
     
         18 . The method of any of  claims 12 - 17 , wherein the lymphangiogenesis inducer comprises a matrix binding domain. 
     
     
         19 . The method of any of  claims 12 - 18 , wherein the lymphangiogenesis inducer comprises a protease cleavage site. 
     
     
         20 . The method of  claim 18  or  19 , wherein the lymphangiogenesis inducer is a modified vascular endothelial growth factor C (VEGF-C) protein comprising a fibrin-binding domain. 
     
     
         21 . The method of  claim 19  or  20 , wherein the protease cleavage site is a matrix metalloprotease cleavage site. 
     
     
         22 . The method of  claim 20  or  21 , wherein the VEGF-C is recombinant, the antigen is recombinant or both the VEGF-C and the antigen are recombinant. 
     
     
         23 . The method of  claim 22 , wherein the VEGF-C, the antigen or both the VEGF-C and the antigen are expressed by a heterologous cell line. 
     
     
         24 . The method of any of  claims 12 - 23 , wherein the matrix or hydrogel is implanted into the subject. 
     
     
         25 . The method of any of  claims 1 - 24 , wherein the lymphangiogenesis inducer and antigen are administered separately. 
     
     
         26 . The method of  claim 25 , wherein the lymphangiogenesis inducer and the antigen are administered up to 1 month apart. 
     
     
         27 . The method of any of  claims 1 - 26 , wherein the subject is a mammal. 
     
     
         28 . The method of any of  claims 1 - 27 , wherein the subject is a human. 
     
     
         29 . The method of any of  claims 1 - 28 , wherein the immune response is a protective immune response. 
     
     
         30 . The method of any of  claims 1 - 29 , wherein the immune response produces antibodies that specifically bind to the antigen or the second antigen. 
     
     
         31 . The method of any of  claims 1 - 30 , wherein the antigen is encoded by a recombinant nucleic acid molecule. 
     
     
         32 . A method of treating or preventing an infection in a subject, the method comprising administering to the subject an isolated lymphangiogenesis inducer and an effective amount of an antigen. 
     
     
         33 . The method of  claim 32 , wherein the lymphangiogenesis inducer is vascular endothelial growth factor C (VEGF-C) or vascular endothelial growth factor D (VEGF-D). 
     
     
         34 . The method of  claim 32  or  33 , wherein the antigen is a bacterial antigen, a viral antigen, a fungal antigen, a helminth antigen or a protozoal antigen. 
     
     
         35 . The method of any of  claims 32 - 34 , wherein the lymphangiogenesis inducer and the antigen are administered in a single composition. 
     
     
         36 . The method of any of  claims 32 - 35 , wherein the lymphangiogenesis inducer and the antigen are provided over multiple administrations. 
     
     
         37 . The method of any of  claims 32 - 36 , wherein the subject is also administered an adjuvant. 
     
     
         38 . The method of any of  claims 32 - 37 , wherein the lymphangiogenesis inducer and the antigen are administered in a single composition comprising an adjuvant. 
     
     
         39 . The method of any of  claims 32 - 38 , wherein the lymphangiogenesis inducer and the antigen are administered in a single composition comprising a pharmaceutically acceptable excipient. 
     
     
         40 . The method of any of  claims 35 - 39 , wherein the composition is administered parenterally, subcutaneously or intramuscularly. 
     
     
         41 . The method of any of  claims 32 - 40 , wherein the subject is administered an effective amount of a second antigen. 
     
     
         42 . The method of any of  claims 32 - 41 , wherein the lymphangiogenesis inducer is incorporated into a matrix. 
     
     
         43 . The method of  claim 42 , wherein the antigen is bound or incorporated into the matrix and is cleavable. 
     
     
         44 . The method of  claims 42 - 43 , wherein the matrix incorporates one or more cleavable chemoattractants or one or more cleavable cytokines. 
     
     
         45 . The method of any one of  claims 42 - 44 , wherein the matrix is a hydrogel. 
     
     
         46 . The method of  claim 45 , wherein the hydrogel is a fibrin hydrogel or a fibrin domain modified polyethylene glycol hydrogel. 
     
     
         47 . The method of any of  claims 42 - 46 , wherein the lymphangiogenesis inducer is capable of binding to the matrix or hydrogel. 
     
     
         48 . The method of any of  claims 42 - 47 , wherein the lymphangiogenesis inducer comprises a matrix binding domain. 
     
     
         49 . The method of any of  claims 42 - 47 , wherein the lymphangiogenesis inducer comprises a protease cleavage site. 
     
     
         50 . The method of  claim 48  or  49 , wherein the lymphangiogenesis inducer is a modified vascular endothelial growth factor C (VEGF-C) protein comprising a fibrin-binding domain. 
     
     
         51 . The method of  claim 49  or  50 , wherein the protease cleavage site is a matrix metalloprotease cleavage site. 
     
     
         52 . The method of  claim 50  or  51 , wherein the VEGF-C is recombinant, the antigen is recombinant or both the VEGF-C and the antigen are recombinant. 
     
     
         53 . The method of  claim 52 , wherein the VEGF-C, the antigen or both the VEGF-C and the antigen are expressed by a heterologous cell line. 
     
     
         54 . The method of any of  claims 42 - 53 , wherein the matrix or hydrogel is implanted into the subject. 
     
     
         55 . The method of any of  claims 32 - 54 , wherein the lymphangiogenesis inducer and antigen are administered separately. 
     
     
         56 . The method of  claim 55 , wherein the lymphangiogenesis inducer and the antigen are administered up to 1 month apart. 
     
     
         57 . The method of any of  claims 32 - 56 , wherein the subject is a mammal. 
     
     
         58 . The method of any of  claims 32 - 57 , wherein the subject is a human. 
     
     
         59 . The method of any of  claims 32 - 58 , wherein administration of the lymphangiogenesis inducer and antigen or second antigen produces antibodies that specifically bind to the antigen or the second antigen. 
     
     
         60 . The method of any of  claims 32 - 59 , wherein the antigen is encoded by a recombinant nucleic acid molecule. 
     
     
         61 . A pharmaceutical composition comprising one or more lymphangiogenesis inducers, an effective amount of an antigen and a pharmaceutically acceptable excipient. 
     
     
         62 . The composition of  claim 61 , wherein the one or more lymphangiogenesis inducer comprise vascular endothelial growth factor C (VEGF-C) or vascular endothelial growth factor D (VEGF-D). 
     
     
         63 . The composition of  claim 61  or  62 , wherein the one or more lymphangiogenesis inducers comprise CCL21. 
     
     
         64 . The composition of any of  claims 61 - 63 , wherein the antigen is a bacterial antigen, a viral antigen, a fungal antigen, a protozoal antigen or a cancer antigen. 
     
     
         65 . The composition of any of  claims 61 - 64 , wherein the composition comprises an adjuvant. 
     
     
         66 . The composition of any of  claims 61 - 65 , wherein the composition is adapted to be administered parenterally, subcutaneously or intramuscularly. 
     
     
         67 . The composition of any of  claims 61 - 66 , wherein the composition comprises an effective amount of a second antigen. 
     
     
         68 . The composition of any of  claims 61 - 67 , wherein the lymphangiogenesis inducer is incorporated into a matrix. 
     
     
         69 . The composition of  claim 68 , wherein the antigen is bound or incorporated into the matrix and is cleavable. 
     
     
         70 . The composition of  claims 68 - 69 , wherein the matrix incorporates one or more cleavable chemoattractants or one or more cleavable cytokines. 
     
     
         71 . The composition of any of  claims 68 - 70 , wherein the matrix is a hydrogel. 
     
     
         72 . The composition of  claim 71 , wherein the hydrogel is a fibrin hydrogel or a fibrin domain modified polyethylene glycol hydrogel. 
     
     
         73 . The composition of any of  claims 68 - 72 , wherein the lymphangiogenesis inducer is bound or incorporated into the matrix or hydrogel. 
     
     
         74 . The composition of any of  claims 68 - 73 , wherein the lymphangiogenesis inducer comprises a matrix binding domain 
     
     
         75 . The composition of  claim 74 , wherein the lymphangiogenesis inducer comprises a protease cleavage site. 
     
     
         76 . The composition of  claim 75 , wherein the lymphangiogenesis inducer is a modified vascular endothelial growth factor C (VEGF-C) protein comprising a fibrin-binding domain and a matrix metalloprotease cleavage site. 
     
     
         77 . The composition of  claim 76 , wherein the VEGF-C is recombinant, the antigen is recombinant or both the VEGF-C and the antigen are recombinant. 
     
     
         78 . The composition of  claim 77 , wherein the VEGF-C, the antigen or both the VEGF-C and the antigen are expressed by a heterologous cell line. 
     
     
         79 . A vaccine comprising the composition of any one of  claims 61 - 78 . 
     
     
         80 . A method of treating neoplasia, dysplasia or cancer, the method comprising administering to a subject one or more lymphangiogenesis inducers and an effective amount of one or more neoplasia, dysplasia or cancer antigens. 
     
     
         81 . The method of  claim 80 , wherein the one or more lymphangiogenesis inducers comprise vascular endothelial growth factor C (VEGF-C) or vascular endothelial growth factor D (VEGF-D). 
     
     
         82 . The method of  claim 80  or  81 , wherein the one or more lymphangiogenesis inducers comprise CCL21. 
     
     
         83 . The method of  claim 80  or  81 , wherein the one or more antigens are cancer antigens. 
     
     
         84 . The method of any of  claims 80 - 83 , wherein the one or more antigens are in a cell lysate. 
     
     
         85 . The method of any of  claims 80 - 84 , wherein the lymphangiogenesis inducer and the one or more neoplasia, dysplasia or cancer antigens are administered in a single composition. 
     
     
         86 . The method of any of  claims 80 - 85 , wherein the lymphangiogenesis inducer and the one or more neoplasia, dysplasia or cancer antigens are provided over multiple administrations. 
     
     
         87 . The method of any of  claims 80 - 86 , wherein the subject is also administered an adjuvant. 
     
     
         88 . The method of any of  claims 80 - 87 , wherein the lymphangiogenesis inducer and the one or more neoplasia, dysplasia or cancer antigens are administered in a single composition comprising an adjuvant. 
     
     
         89 . The method of any of  claims 80 - 88 , wherein the lymphangiogenesis inducer and the one or more neoplasia, dysplasia or cancer antigens are administered in a single composition comprising a pharmaceutically acceptable excipient. 
     
     
         90 . The method of any of  claims 85 - 89 , wherein the composition is administered parenterally, subcutaneously or intramuscularly. 
     
     
         91 . The method of any of  claims 80 - 90 , wherein the subject is administered an effective amount of a second neoplasia, dysplasia or cancer antigen. 
     
     
         92 . The method of any of  claims 80 - 91 , wherein the lymphangiogenesis inducer is incorporated into a matrix. 
     
     
         93 . The method of  claim 92 , wherein the antigen is bound or incorporated into the matrix and is cleavable. 
     
     
         94 . The method of  claims 92 - 93 , wherein the matrix incorporates one or more cleavable chemoattractants or one or more cleavable cytokines. 
     
     
         95 . The method of any one of  claims 92 - 94 , wherein the matrix is a hydrogel. 
     
     
         96 . The method of  claim 95 , wherein the hydrogel is a fibrin hydrogel or a fibrin domain modified polyethylene glycol hydrogel. 
     
     
         97 . The method of any of  claims 92 - 96 , wherein the lymphangiogenesis inducer is capable of binding to the matrix or hydrogel. 
     
     
         98 . The method of any of  claims 92 - 97 , wherein the lymphangiogenesis inducer comprises a matrix binding domain. 
     
     
         99 . The method of any of  claims 92 - 98 , wherein the lymphangiogenesis inducer comprises a protease cleavage site. 
     
     
         100 . The method of  claim 98 , wherein the lymphangiogenesis inducer is a modified vascular endothelial growth factor C (VEGF-C) protein comprising a fibrin-binding domain. 
     
     
         101 . The method of  claim 99  or  100 , wherein the lymphangiogenesis inducer is a modified vascular endothelial growth factor C (VEGF-C) protein comprising a matrix metalloprotease cleavage site. 
     
     
         102 . The method of  claim 100  or  101 , wherein the VEGF-C is recombinant, the one or more neoplasia, dysplasia or cancer antigens are recombinant or both the VEGF-C and the one or more antigens are recombinant. 
     
     
         103 . The method of  claim 102 , wherein the VEGF-C, the antigen or both the VEGF-C and the one or more neoplasia, dysplasia or cancer antigens are expressed by a heterologous cell line. 
     
     
         104 . The method of any of  claims 80 - 101 , wherein the matrix or hydrogel is implanted into the subject. 
     
     
         105 . The method of any of  claims 80 - 104 , wherein the lymphangiogenesis inducer and one or more neoplasia, dysplasia or cancer antigens are administered separately. 
     
     
         106 . The method of  claim 105 , wherein the lymphangiogenesis inducer and one or more neoplasia, dysplasia or cancer antigens are administered up to 1 month apart. 
     
     
         107 . The method of any of  claims 80 - 106 , wherein the subject is a mammal. 
     
     
         108 . The method of any of  claims 80 - 107 , wherein the subject is a human. 
     
     
         109 . The method of any of  claims 80 - 108 , wherein administration of the lymphangiogenesis inducer and the one or more neoplasia, dysplasia or cancer antigens or the second neoplasia, dysplasia or cancer antigen produces antibodies that specifically bind to the one or more neoplasia, dysplasia or cancer antigens or to the second neoplasia, dysplasia or cancer antigen. 
     
     
         110 . The method of any of  claims 80 - 109 , wherein the one or more neoplasia, dysplasia or cancer antigens are encoded by a recombinant nucleic acid molecule. 
     
     
         111 . The method of any one of  claims 80 - 110 , wherein the cancer comprises breast cancer or melanoma or a neoplasia or dysplasia of the breast or skin. 
     
     
         112 . The method of any one of  claims 80 - 111 , wherein the method further comprises administration of an immunotherapy. 
     
     
         113 . The method of  claim 112 , wherein the immunotherapy comprises a checkpoint blockade inhibitor, adoptive T cell therapy, a chimeric antigen receptor, a STING agonist, cytolytic virus therapy, one or more additional antigens, tumor cell lysate, tolerance-breaking peptide antigen, a dendritic cell vaccine, or an antibody-antigen conjugate. 
     
     
         114 . A method for treating a cancer patient with cancer immunotherapy comprising administering cancer immunotherapy to the patient after detecting in a serum sample from that patient a level of VEGF-C and/or CCL21 that is indicative of response to cancer immunotherapy. 
     
     
         115 . The method of  claim 114 , further comprising comparing the level of VEGF-C and/or CCL21 to a control level. 
     
     
         116 . The method of  claim 115 , wherein the control level is a level of VEGF-C or CCL21 expression from serum of patients who affirmatively respond to the cancer immunotherapy. 
     
     
         117 . The method of  claim 116 , wherein patients who affirmatively respond to the cancer immunotherapy exhibit at least a 25% reduction in tumor growth following treatment with the cancer immunotherapy. 
     
     
         118 . The method of  claim 115 , wherein the control level is a level of VEGF-C or CCL21 expression that is the median level in serum of cancer patients. 
     
     
         119 . The method of  claim 118 , wherein the patient is administered immunotherapy after being determined to have a level of expression of VEGF-C or CCL21 that is increased as compared to the level of expression in serum from noncancer patients. 
     
     
         120 . The method of  claim 115 , wherein the control level is a level or range of level of VEGF-C or CCL21 expression from serum of patients who do not respond affirmatively to the cancer immunotherapy. 
     
     
         121 . The method of  claim 115 , wherein the control level is a level or range of level of VEGF-C or CCL21 expression in serum from patients who do not respond affirmatively to the cancer immunotherapy. 
     
     
         122 . The method of any of  claims 114 - 121 , further comprising obtaining serum from the patient. 
     
     
         123 . The method of any of  claims 114 - 122 , further comprising measuring the level of VEGF-C and/or CCL21 expression in serum from the patient. 
     
     
         124 . The method of  claim 123 , wherein measuring the level of VEGF-C and/or CCL21 expression comprises using a peptide or polypeptide that binds VEGF-C and/or CCL21. 
     
     
         125 . The method of any of  claims 114 - 124 , wherein the immunotherapy is a checkpoint blockade inhibitor, adoptive T cell therapy, a chimeric antigen receptor, a STING agonist, cytolytic virus therapy, one or more additional antigens, tumor cell lysate, tolerance-breaking peptide antigen, a dendritic cell vaccine, or an antibody-antigen conjugate. 
     
     
         126 . A method for predicting the efficacy of cancer immunotherapy in a patient comprising:
 measuring a level of VEGF-C and/or CCL21 protein expression in a serum sample from the patient; and,   comparing the level of VEGF-C and/or CCL21 protein expression to a level in a control sample.   
     
     
         127 . The method of  claim 126 , further comprising predicting efficacy of the cancer immunotherapy in the patient if the level of VEGF-C and/or CCL21 protein expression is increased compared to the level of VEGF-C and/or CCL21 protein expression in a patient without cancer.