Endogenous tgf-beta inhibited dendritic cells

Abstract

Disclosed are methods and composition for treating cancer in a subject using TGF-beta inhibited dendritic cells.

Description

[0001] ENDOGENOUS TGF-BETA INHIBITED DENDRITIC CELLS

[0002] I. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0003] This invention was made with Government Support under Grant No. P01CA234212 awarded by the National Institutes of Health. The Government has certain right in the invention.

[0004] II. CROSS REFERENCE TO RELATED APPLICATIONS

[0005] This application claims the benefit of U.S. Provisional Application No. 63 / 594,093, filed on October 30, 2023, which is incorporated herein by reference in its entirety.

[0006] III. BACKGROUND

[0007] 38. Dendritic cells can induce effective cancer immunity in experimental animals and cancer patients, leading to their applications as cancer therapies. Several groups of factors are needed for their clinical activity: a) ability to present antigens (“signal 1”); b) ability to provide costimulatory factors (“signal 2”); c) ability to migrate to the draining lymph nodes and d) ability to produce IL- 12 during contact with antigen specific T cells (“signal 3”), are differentially regulated at different stages of DC maturation, and limited by the maturation-associated DC “exhaustion”, making it difficult to generate DC product optimized for all these features.

[0008] 39. The past efforts to enhance and prolong the duration of the Thl, NK, and CTL activating (and attracting) effector functions, let to the development of several “type- 1 -polarized” DC populations, induced by viral mimics, activated CD8+ T cells and NK cells, which produce much higher levels of IL-12p70, the factor need for the clinical benefit of DC therapies, and other desirable effector cell-attracting cytokines and chemokines (IL-18, CCL5, CCL19, CXCL10, CXCL11), but lower levels of “suppressor / regulatory” factors, such as CCL22. Such polarized DCs proved superior to multiple forms of non-polarized DCs in induction of antitumor Thl, NK, and CTLs in preclinical in vitro and in vivo models and were shown capable of inducing clinical responses in patients with advanced cancer.

[0009] 40. However, the polarized DCs still undergo eventual “exhaustion”. Due to these limitations, there is a need to further improve the process of generation of polarized DCs and other DCs to use in cancer therapy.

[0010] IV. SUMMARY

[0011] 41. Disclosed herein are methods and compositions related to treatment of cancer using ex-vivo generated endogenous TGF-P inhibited antigen-presenting cells. 42. Disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, hut not limited to checkpoint-resistant metastatic melanoma)) in a subject, comprising activating a population of antigen specific immune cells (such as, for example, CD4 T cells (including, but not limited to Thl helper cells), CD8 T cells (including, but not limited to cytotoxic T cells such as, for example, effector CD8 T cells, and central memory CD8 T cells), natural killer (NK) cells, and / or NK T cells) in vitro by co-culturing with a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage) loaded ex vivo with at least one or more tumor antigen peptides, thereby obtaining activated antigen specific immune cells; and administering to the subject an effective amount of the activated antigen specific immune cells, wherein the population of antigen specific immune cells are activated prior to administration. In some aspects, the antigen-presenting cells are loaded with at least one or more tumor antigen peptides for about 4 days to about 21 days. In some aspects, the tumor antigen peptide can be 60, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 amino acids long. In some aspects, the tumor antigen peptide can comprise an MHC-1 or MHC-II epitope. In some aspects, the co-culturing of the antigen specific immune cell and the antigen presenting cell occurs ex vivo or in vitro.

[0012] 43. Also disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein the one or more DC are matured in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL- 1 P), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), and polyinosinic:poly cytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6). In some aspects, the method further comprises addition of inhibitors of transforming growth factor beta (TGF-P) (such as, for example, LY364947, SD208 or SB431542) signaling to pre-maturation culture or the maturation culture.

[0013] 44. In one aspect, disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein aDCl produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

[0014] 45. Also disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, but not limited to checkpoint-resistant metastatic melanoma)) in a subject, comprising administering to the subject an effective amount of a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type-1 polarized dendritic cell (aDCl) B cell, or macrophage), loaded with at least one or more tumor antigen peptides. In some aspects, the tumor antigen peptide can be 60, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 amino acids long. In some aspects, the tumor antigen peptide can comprise an MHC-1 or MHC-II epitope.

[0015] 46. Also disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein the one or more DC are matured in presence of a maturation culture, wherein the maturation culture comprises interleukin- 10 (IL-10), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), and polyinosinic:poly cytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6). In some aspects, the method further comprises addition of inhibitors of transforming growth factor beta (TGF-0) (such as, for example, LY364947, SD208 or SB431542) signaling to pre-maturation culture or the maturation culture.

[0016] 47. In one aspect, disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein aDCl produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

[0017] 48. In one aspect, disclosed herein are cancer vaccines comprising a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage) loaded ex vivo with at least one or more tumor antigen peptides. In some aspects, the tumor antigen peptide can be 60, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 amino acids long. In some aspects, the tumor antigen peptide can comprise an MHC-1 or MHC-1I epitope. In some aspects the tumor antigen peptide can be an autologous or allogeneic peptide.

[0018] 49. Also disclosed herein are cancer vaccines of any preceding aspect, wherein the population of antigen-presenting cells comprise inhibited endogenous transforming growth factor beta (TGF-P) signaling.

[0019] 50. In one aspect disclosed herein are cancer vaccines of any preceding aspect, wherein the antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage) with inhibited endogenous TGF-P signaling are matured in presence of a maturation culture. In some aspects, the maturation culture comprises interleukin- ip (IL-ip), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), a TLR3 ligand (such as, for example, Polyinosinic:polycytidylic acid (poly- I:C)), prostaglandin E2 (PGE2), and / or interleukin-6 (IL-6). In some aspects, antigen preceding cells are further cultured with TGF-P signaling inhibitors (such as, for example, LY364947, SD208 or SB431542) in the pre-maturation culture or the maturation culture.

[0020] 51. Also disclosed herein are cancer vaccines of any preceding aspect, wherein the cancer vaccine produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

[0021] 52. In one aspect, disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, but not limited to checkpoint-resistant metastatic melanoma)) in a subject using a population of activated antigen specific immune cells, comprising a) obtaining a population of peripheral blood mononuclear cells (PBMCs) from the subject; b) obtaining a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage) and a population of immune cells (such as, for example, CD4 T cells (including, but not limited to Thl helper cells), CD8 T cells (including, but not limited to cytotoxic T cells such as, for example, effector CD8 T cells, and central memory CD8 T cells), natural killer (NK) cells, and / or NK T cells) from the population of PBMCs; c) inducing maturation of the population of antigen-presenting cells, thereby obtaining mature antigen-presenting cells; d) blocking endogenous TGF-P signaling in the mature antigen-presenting cells, thereby obtaining TGF-P inhibited antigen-presenting cells; e) contacting the TGF-P inhibited antigen-presenting cells with at least one or more tumor antigen peptides to obtain antigen loaded TGF-P inhibited antigen-presenting cells; f) co-culturing the antigen loaded TGF-P inhibited antigen-presenting cells with the population of immune cells to obtain the population of activated antigen specific immune cells (such as, for example, co-culturing for about 4 days to about 21 days); and g) administering to the subject an effective amount of the population of activated antigen specific immune cells. In some aspects, the tumor antigen peptide can be 60, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 amino acids long. In some aspects, the tumor antigen peptide can comprise an MHC-1 or MHC-II epitope. In some aspects, the population of activated antigen specific immune cells are administered to the subject at least three times. In some aspects, the interval between each administration of the population of activated antigen specific immune cells is about 0.5 month to about 2 months.

[0022] 53. Also disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein the one or more DC are matured in presence of a maturation culture. In some aspects, the maturation culture comprises interleukin- ip (IL- 1 P), tumor necrosis factor-a (TNFa), interferon- a (IFNa), interferon-y (IFNy), a TLR3 ligand (such as, for example, Polyinosinic:polycytidylic acid (poly-I:C)), prostaglandin E2 (PGE2), and / or interleukin-6 (IL-6).

[0023] 54. In one aspect, disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein step d further comprises addition of inhibitors of transforming growth factor beta (TGF- P) signaling (such as, for example, LY364947, SD208 or SB431542) to the culture before or during the maturation culture.

[0024] 55. Also disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis of any preceding aspect, wherein aDCl produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

[0025] 56. In one aspect disclosed herein is an ex vivo -generated antigen-presenting cell, obtained by the method comprising culturing ex vzvo-generated antigen-presenting cell (including, but not limited to dendritic cell (DC), B cell, or macrophage) in the presence of inhibitors of transforming growth factor beta (TGF-P) signaling (such as, for example, small molecule, protein or RNA blocker of the TGF-P signaling or TGF-P production including, but not limited to LY364947, SD208 or SB431542); and adding the inhibitors of TGF-P signaling to a pre-maturation culture or a maturation culture. In some aspects, the ex vzvo-generated antigen- presenting cell comprises alpha- type- 1 polarized dendritic cell (aDCl) exposed ex vivo to a combination of type-I and type-II interferons (IFNs); wherein the type-I IFNs comprise interferon-a (IFNa) or interferon-P (IFNP) and type-II IFNs comprise interferon-y (IFNy). In some aspects, the aDCl is activated in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL-ip), tumor necrosis factor-a (TNFa), IFNa, IFNy, and a TLR3 ligand (such as, for example, polyinosinic:poly cytidylic acid (poly-I:C)

[0026] 57. Also disclosed herein is an ex vzvo-generated antigen-presenting cell of any preceding aspect, wherein the Interleukin ip (ILip) is at a concentration of about 0.2-200 ng / ml, the Tumor necrosis factor a (TNFa) is at a concentration of about 0.2-200 ng / ml, the Interferon a (IFNa) is at a concentration of about 1-10,000 U / ml, the Interferon y (IFNy) is at a concentration of about 1-10,000 U / ml and / or the TLR3 ligand (such as for example, Polyinosinic:poly cytidylic acid (poly-I:C)) is at a concentration of 0.2 ng / ml - 500 pg / ml.

[0027] 58. In one aspect, disclosed herein is an ex vzvo-generated antigen-presenting cell of any preceding aspect, wherein the said ex vzvo-generated antigen-presenting cell produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 9 (CXCL9, )C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response to CD40 ligand (CD40L) stimulation as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-P signaling.

[0028] 59. Also disclosed herein is an ex rzTo-generated antigen-presenting cell of any preceding aspect, wherein the said ex vi vo-generated antigen-presenting cell produces at least 50% or more of IL-12p70, IL-18, CXCL9, CXCL10, CXCL11, CCL5, or CCL19 in response after 24 hours of stimulation to CD40L as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-P signaling.

[0029] 60. In one aspect disclosed herein is a cancer vaccine comprising the ex vzvo-generated antigen-presenting cell of any preceding aspect loaded ex vivo with at least one or more tumor antigen peptides.

[0030] 61. Also disclosed herein are activated antigen specific T-cell, wherein the T cell is activated with the an ex vzvo-generated antigen-presenting cell of any preceding aspect.

[0031] V. BRIEF DESCRIPTION OF THE FIGURES

[0032] 27. FIGS. 1 A-1C show clinical activity of aDCl vaccines in patients with brain cancer and PDl-non-responsive metastatic melanoma. FIG. 1A from the NCT00766753 clinical trial shows complete radiologic response to vaccination with aDCls loaded glioma stem-cells- targeting antigenic peptides in a patient with recurrent glioma (MRI). Objective responses (all long-lasting) were observed in a total of 4 / 19 patients. FIG. IB shows correlation between IL- 12p70 production by aDCl vaccines of the individual patients (in vitro', after 24h CD40L stimulation; ELISA) and the patients’ progression-free survival (PFS: red dotted lines represented 2-4 months PFS expected in this patient group). FIG. 1 C from NCT01876212 clinical trial shows vaccination aDCls loaded with tumor blood vessel antigenic (TBVA) peptides. Example of a near-compete response in a patient with metastatic melanoma who previously progressed on PD1 blockade and received. So far, 4 objective responses (2 nCRs, 2 PR) and 2 SDs (>9m) were seen in 12 pts with PD1 -resistant metastatic melanoma. All patients who responded or stabilized developed CTL responses against TBVA but also against melanoma-lineage specific CTL epitopes not included in the vaccine (epitope spreading).

[0033] 28. FIGS. 2A-2B show endogenous TGF-pi levels are high in immature DC, fully blocked in during PGE2-assisted maturation of “standard” DCs (sDC), but only partially downregulated upon DC maturation in type- 1 -polarizing conditions (aDCl; matured in IL-ip, TNFa, IFNy, IFNa and poly-LC). TGF-pi transcript levels analyzed by qRT-PCR. Note the complete downregulation in sDC (FIG. 2A), but only partial downregulation in aDCl (FIG. 2B) progressively seen at 24 and 48 hr post maturation.

[0034] 29. FIGS. 3A-3F show Inhibition of TGF-P during DC maturation enhances and prolongs the ability of resulting aDCls to produce CTL-attracting and CTL-activating factors. Human monocyte-derived DCs were matured in aDCl cocktail (IL-ip, TNFa, IFNy, IFNa and poly-I:C) with or without TGF-P-R1 inhibitors LY364947, SD208 or SB431542. FIGS 3 A and 3B show that after maturation, aDCls were harvested at indicated timepoints and re-plated with soluble (s)CD40L to induce the secretion of CTL / Thl / NK-activating cytokine IL-12p70 (FIG. 3A) and CTL / Thl / NK-attracting 1 chemokine CCL5 (FIG. 3B). FIGS. 3C, 3D, 3E and 3F show that aDCls were harvested 24 hrs after maturation and replated with sCD40L induce IL-12p70 (FIG. 3C and FIG.3E) , CCL5 secretion (FIG. 3D) or, CCL19,the attractant for naive and memory T cells (CTL precursors) and additional mature DCs. FIG. 3E shows advantage of prolonged pretreatment of immature DCs to TGF-P blockade (pretreated with LY364947 for 96hrs before maturation). After 24 hours of maturation and (when indicated) 24 hours of sDC40L stimulation, supernatants (FIGS. 3 A, 3B, 3C, 3D and 3E) or mRNA (FIG. 3F) were collected and analyzed for protein secretion by ELISA or CCL19 expression levels by quantitative RT-PCR.

[0035] 30. FIGS. 4A-4B show desirable selectivity of action of TGF-P blockade: Inhibition of TGF-P signaling during DC maturation does not enhance the production aDCl produced suppressive factors. Human monocyte-derived DCs were matured with aDCl cocktail (IL- 1|3, TNFa, IFNy, IFNa and poly-I:C) in the absence or presence of TGF-P-R1 inhibitor LY364947. Levels of CCL22, a Treg-attracting chemokine (FIG. 4A) and ID01, an immunosuppressive factor (FIG. 4B) were analyzed by qRT-PCR in immature DCs (iDC) or aDCls 24hr post maturation.

[0036] 31 . FIGS. 5A-5B show DCs generated in the presence of TGF-beta blocker (small molecule SB431542) show enhanced ability to induce expansion of MART- 1- specific CTLs with improved ability to specifically recognize MART-1 expressing melanoma cell lines and to respond with secretion of IFNy. Original aDCls or TGF-0 blocked aDCls generated from HLA- A2 positive donor, were loaded with MART-1 antigen and used in invitro sensitization cultures with autologous CD8+ T cells. After expansion, the presence of antigen- specific T cells (FIG.

[0037] 5 A; MART-1 tetramer staining) and their ability to secrete IFNy in response to activation with MART-1 expressing melanoma cell lines (MEL-526, MEL-624, 2183-HER4) or control MART-1 negative SW620 cancer cells (FIG. 5B; IFNy ELISPOT).

[0038] VI. DETAILED DESCRIPTION

[0039] 32. Before the present compounds, compositions, articles, devices, and / or methods are disclosed and described, it is to be understood that they are not limited to specific synthetic methods or specific recombinant biotechnology methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0040] A. Definitions

[0041] 33. As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a pharmaceutical carrier” includes mixtures of two or more such carriers, and the like.

[0042] 34. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “10” is disclosed the “less than or equal to 10” as well as “greater than or equal to 10” is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point 15 are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0043] 35. The term “subject” is defined herein to include animals such as mammals, including, but not limited to, primates (e.g., humans), cows, horses, pigs, sheep, goats, dogs, cats, rabbits, rats, mice and the like. In some embodiments, the subject is a human.

[0044] 36. “Administration” to a subject includes any route of introducing or delivering to a subject an agent. Administration can be carried out by any suitable route, including oral, topical, intravenous, subcutaneous, transcutaneous, transdermal, intramuscular, intra-joint, parenteral, intra-arteriole, intradermal, intraventricular, intracranial, intraperitoneal, intralesional, intranasal, rectal, vaginal, by inhalation, via an implanted reservoir, parenteral (e.g., subcutaneous, intravenous, intramuscular, intra- articular, intra-synovial, intrasternal, intrathecal, intraperitoneal, intrahepatic, intralesional, and intracranial injections or infusion techniques), and the like. “Systemic administration” refers to the introducing or delivering to a subject an agent via a route which introduces or delivers the agent to extensive areas of the subject’s body (e.g. greater than 50% of the body), for example through entrance into the circulatory or lymph systems. By contrast, “local administration” refers to the introducing or delivery to a subject an agent via a route which introduces or delivers the agent to the area or area immediately adjacent to the point of administration and does not introduce the agent systemically in a therapeutically significant amount. For example, locally administered agents are easily detectable in the local vicinity of the point of administration but are undetectable or detectable at negligible amounts in distal parts of the subject’s body. Administration includes self-administration and the administration by another. 37. "Biocompatible" generally refers to a material and any metabolites or degradation products thereof that are generally non-toxic to the recipient and do not cause significant adverse effects to the subject.

[0045] 38. "Comprising" is intended to mean that the compositions, methods, etc. include the recited elements, but do not exclude others. "Consisting essentially of’ when used to define compositions and methods, shall mean including the recited elements, but excluding other elements of any essential significance to the combination. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants from the isolation and purification method and pharmaceutically acceptable carriers, such as phosphate buffered saline, preservatives, and the like. "Consisting of” shall mean excluding more than trace elements of other ingredients and substantial method steps for administering the compositions of this invention. Embodiments defined by each of these transition terms are within the scope of this invention.

[0046] 39. A “control” is an alternative subject or sample used in an experiment for comparison purposes. A control can be "positive" or "negative."

[0047] 40. “Controlled release” or “sustained release” refers to release of an agent from a given dosage form in a controlled fashion in order to achieve the desired pharmacokinetic profile in vivo. An aspect of “controlled release” agent delivery is the ability to manipulate the formulation and / or dosage form in order to establish the desired kinetics of agent release.

[0048] 41. “Effective amount” of an agent refers to a sufficient amount of an agent to provide a desired effect. The amount of agent that is “effective” will vary from subject to subject, depending on many factors such as the age and general condition of the subject, the particular agent or agents, and the like. Thus, it is not always possible to specify a quantified “effective amount.” However, an appropriate “effective amount” in any subject case may be determined by one of ordinary skill in the art using routine experimentation. Also, as used herein, and unless specifically stated otherwise, an “effective amount” of an agent can also refer to an amount covering both therapeutically effective amounts and prophylactically effective amounts. An “effective amount” of an agent necessary to achieve a therapeutic effect may vary according to factors such as the age, sex, and weight of the subject. Dosage regimens can be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.

[0049] 42. A "decrease" can refer to any change that results in a smaller gene expression, protein production, amount of a symptom, disease, composition, condition, or activity. A substance is also understood to decrease the genetic output of a gene when the genetic output of the gene product with the substance is less relative to the output of the gene product without the substance. Also, for example, a decrease can be a change in the symptoms of a disorder such that the symptoms are less than previously observed. A decrease can be any individual, median, or average decrease in a condition, symptom, activity, composition in a statistically significant amount. Thus, the decrease can be a 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% decrease so long as the decrease is statistically significant.

[0050] 43. "Inhibit," "inhibiting," and "inhibition" mean to decrease an activity, response, condition, disease, or other biological parameter. This can include but is not limited to the complete ablation of the activity, response, condition, or disease. This may also include, for example, a 10% reduction in the activity, response, condition, or disease as compared to the native or control level. Thus, the reduction can be a 10, 20, 30, 40, 50, 60, 70, 80, 90, 100%, or any amount of reduction in between as compared to native or control levels.

[0051] 44. The terms “prevent,” “preventing,” “prevention,” and grammatical variations thereof as used herein, refer to a method of partially or completely delaying or precluding the onset or recurrence of a disease and / or one or more of its attendant symptoms or barring a subject from acquiring or reacquiring a disease or reducing a subject’s risk of acquiring or reacquiring a disease or one or more of its attendant symptoms.

[0052] 45. "Pharmaceutically acceptable" component can refer to a component that is not biologically or otherwise undesirable, i.e., the component may be incorporated into a pharmaceutical formulation of the invention and administered to a subject as described herein without causing significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the formulation in which it is contained. When used in reference to administration to a human, the term generally implies the component has met the required standards of toxicological and manufacturing testing or that it is included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug Administration.

[0053] 46. "Pharmaceutically acceptable carrier" (sometimes referred to as a “carrier”) means a carrier or excipient that is useful in preparing a pharmaceutical or therapeutic composition that is generally safe and non-toxic and includes a carrier that is acceptable for veterinary and / or human pharmaceutical or therapeutic use. The terms "carrier" or "pharmaceutically acceptable carrier" can include, but are not limited to, phosphate buffered saline solution, water, emulsions (such as an oil / water or water / oil emulsion) and / or various types of wetting agents. As used herein, the term "carrier" encompasses, but is not limited to, any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer, or other material well known in the art for use in pharmaceutical formulations and as described further herein.

[0054] 47. “Pharmacologically active” (or simply “active”), as in a “pharmacologically active” derivative or analog, can refer to a derivative or analog (e.g., a salt, ester, amide, conjugate, metabolite, isomer, fragment, etc.) having the same type of pharmacological activity as the parent compound and approximately equivalent in degree.

[0055] 48. “Therapeutic agent” refers to any composition that has a beneficial biological effect. Beneficial biological effects include both therapeutic effects, e.g., treatment of a disorder or other undesirable physiological condition, and prophylactic effects, e.g., prevention of a disorder or other undesirable physiological condition (e.g., a non-immunogenic cancer). The terms also encompass pharmaceutically acceptable, pharmacologically active derivatives of beneficial agents specifically mentioned herein, including, but not limited to, salts, esters, amides, proagents, active metabolites, isomers, fragments, analogs, and the like. When the terms “therapeutic agent” is used, then, or when a particular agent is specifically identified, it is to be understood that the term includes the agent per se as well as pharmaceutically acceptable, pharmacologically active salts, esters, amides, proagents, conjugates, active metabolites, isomers, fragments, analogs, etc.

[0056] 49. “Therapeutically effective amount” or “therapeutically effective dose” of a composition (e.g. a composition comprising an agent) refers to an amount that is effective to achieve a desired therapeutic result. In some embodiments, a desired therapeutic result is the control of type I diabetes. In some embodiments, a desired therapeutic result is the control of obesity. Therapeutically effective amounts of a given therapeutic agent will typically vary with respect to factors such as the type and severity of the disorder or disease being treated and the age, gender, and weight of the subject. The term can also refer to an amount of a therapeutic agent, or a rate of delivery of a therapeutic agent (e.g., amount over time), effective to facilitate a desired therapeutic effect, such as pain (i.e., nociception) relief. The precise desired therapeutic effect will vary according to the condition to be treated, the tolerance of the subject, the agent and / or agent formulation to be administered (e.g., the potency of the therapeutic agent, the concentration of agent in the formulation, and the like), and a variety of other factors that are appreciated by those of ordinary skill in the art. In some instances, a desired biological or medical response is achieved following administration of multiple dosages of the composition to the subject over a period of days, weeks, or years.

[0057] 50. In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings: 51. “Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0058] 52. Throughout this application, various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this pertains. The references disclosed are also individually and specifically incorporated by reference herein for the material contained in them that is discussed in the sentence in which the reference is relied upon.

[0059] Dendritic cells (DC) in cancer immunity

[0060] 53. Dendritic cells are key inducers of immunity. DCs activate the lymph node-based naive and central memory T cells with MHC / peptide complexes (so-called “signal 1”) and costimulatory molecules (“signal 2”) inducing the expansion of T cells and their effector functions (cytokine production, cytotoxicity, migration to sites of inflammation). In addition, DCs can instruct naive CD4+T cells to differentiate into Thl, Th2, or T regulatory- type effector / memory cells (“signal 3), and differentially promote the development of either type-1, cell-mediated immunity, or type-2 humoral responses. Type-1 Th (Thl) T cells, most desirable in cancer, can directly kill tumor cells and provide CD40L-mediated helper signals for CTL development and maintenance, as well as for the optimal NK cell activity. Induction of Thl cells is promoted by several DC-produced cytokines, particularly by IL-12p70. IL-12p70 also provides essential signals for the activation, effector functions, and survival of NK cells and cytotoxic T lymphocytes.

[0061] Methods of adoptive cell therapy in treating cancer

[0062] 54. Disclosed herein are methods and compositions related to treatment of cancer using ex-vivo generated endogenous TGF-P inhibited antigen-presenting cells. In some embodiments, the cancer includes, but is not limited to acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma), appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g., cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast), brain cancer (e.g., meningioma; glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchus cancer, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., adenocarcinoma of the esophagus, Barrett’s adenocarinoma), Ewing's sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma), familiar hypereosinophilia, gall bladder cancer, gastric cancer (e.g., stomach adenocarcinoma), gastrointestinal stromal tumor (GIST), head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma (OSCC), throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)), hematopoietic cancers (e.g., leukemia such as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T- cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., “Waldenstrom’s macroglobulinemia”), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B -lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); a mixture of one or more leukemia / lymphoma as described above; and multiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease), hemangioblastoma, inflammatory myofibroblastic tumors, immunocytic amyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms’ tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g., gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), Islet cell tumors), penile cancer (e.g., Paget's disease of the penis and scrotum), pinealoma, primitive neuroectodermal tumor (PNT), prostate cancer (e.g., prostate adenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g., appendix cancer), soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland carcinoma, sweat gland carcinoma, synovioma, testicular cancer (e.g., seminoma, testicular embryonal carcinoma), thyroid cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer), urethral cancer, vaginal cancer and vulvar cancer (e.g., Paget's disease of the vulva).

[0063] 55. In one aspect, disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, but not limited to checkpoint-resistant metastatic melanoma)) in a subject comprising activating a population of antigen specific immune cells (such as, for example, CD4 T cells (including, but not limited to Thl helper cells), CD8 T cells (including, but not limited to cytotoxic T cells such as, for example, effector CD8 T cells, and central memory CD8 T cells), natural killer (NK) cells, and / or NK T cells) in vitro by co-culturing with a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage) loaded ex vivo with at least one or more tumor antigen peptides, thereby obtaining activated antigen specific immune cells; and administering to the subject an effective amount of the activated antigen specific immune cells, wherein the population of antigen specific immune cells are activated prior to administration.

[0064] 56. Also disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, but not limited to checkpoint-resistant metastatic melanoma)) in a subject, comprising administering to the subject an effective amount of a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage), loaded with at least one or more tumor antigen peptides.

[0065] 57. In one aspect, disclosed herein are methods of treating, inhibiting, decreasing, reducing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, but not limited to checkpoint-resistant metastatic melanoma)) in a subject using a population of activated antigen specific immune cells, comprising a) obtaining a population of peripheral blood mononuclear cells (PBMCs) from the subject; b) obtaining a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type-1 polarized dendritic cell (aDCl) B cell, or macrophage) and a population of immune cells (such as, for example, CD4 T cells (including, but not limited to Thl helper cells), CD 8 T cells (including, but not limited to cytotoxic T cells such as, for example, effector CD8 T cells, and central memory CD8 T cells), natural killer (NK) cells, and / or NK T cells) from the population of PBMCs; c) inducing maturation of the population of antigen-presenting cells, thereby obtaining mature antigen-presenting cells; d) blocking endogenous TGF-0 signaling in the mature antigen-presenting cells, thereby obtaining TGF-P inhibited antigen-presenting cells; e) contacting the TGF- inhibited antigen-presenting cells with at least one or more tumor antigen peptides to obtain antigen loaded TGF-P inhibited antigen-presenting cells; f) co-culturing the antigen loaded TGF-P inhibited antigen-presenting cells with the population of immune cells to obtain the population of activated antigen specific immune cells (such as, for example, co-culturing for about 4 days to about 21 days); and g) administering to the subject an effective amount of the population of activated antigen specific immune cells.

[0066] 58. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 1 day. In some embodiments, the population of antigen specific immune cells is cocultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 2 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 3 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 4 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen- presenting cells loaded with at least one or more tumor antigen peptides for about 5 days, n some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 6 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 7 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 8 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 9 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen- presenting cells loaded with at least one or more tumor antigen peptides for about 10 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 11 days. In some embodiments, the population of antigen specific immune cells is co- cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 12 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 13 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 14 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 15 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 16 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 17 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 18 days. In some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen- presenting cells loaded with at least one or more tumor antigen peptides for about 19 days. Tn some embodiments, the population of antigen specific immune cells is co-cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 20 days. In some embodiments, the population of antigen specific immune cells is co- cultured with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 21 days.

[0067] 59. In some embodiments, disclosed herein, the tumor antigen peptides are about 20 to about 40 amino acids long. In some embodiments, the tumor antigen peptides is about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 amino acids long. In some embodiments, the tumor antigen peptides is about 2 amino acids long. In some embodiments, the tumor antigen peptides is about 3 amino acids long. In some embodiments, the tumor antigen peptides is about 4 amino acids long. In some embodiments, the tumor antigen peptides is about 5 amino acids long. In some embodiments, the tumor antigen peptides is about 6 amino acids long. In some embodiments, the tumor antigen peptides is about 7 amino acids long. In some embodiments, the tumor antigen peptides is about 8 amino acids long. In some embodiments, the tumor antigen peptides is about 9 amino acids long. In some embodiments, the tumor antigen peptides is about 10 amino acids long. In some embodiments, the tumor antigen peptides is about 11 amino acids long. In some embodiments, the tumor antigen peptides is about 12 amino acids long. In some embodiments, the tumor antigen peptides is about 13 amino acids long. In some embodiments, the tumor antigen peptides is about 14 amino acids long. In some embodiments, the tumor antigen peptides is about 15 amino acids long. In some embodiments, the tumor antigen peptides is about 16 amino acids long. In some embodiments, the tumor antigen peptides is about 17 amino acids long. In some embodiments, the tumor antigen peptides is about 18 amino acids long. In some embodiments, the tumor antigen peptides is about 19 amino acids long. In some embodiments, the tumor antigen peptides is about 20 amino acids long. In some embodiments, the tumor antigen peptides is about 21 amino acids long. In some embodiments, the tumor antigen peptides is about 22 amino acids long. In some embodiments, the tumor antigen peptides is about 23 amino acids long. In some embodiments, the tumor antigen peptides is about 24 amino acids long. In some embodiments, the tumor antigen peptides is about 25 amino acids long. In some embodiments, the tumor antigen peptides is about 26 amino acids long. In some embodiments, the tumor antigen peptides is about 27 amino acids long. In some embodiments, the tumor antigen peptides is about 28 amino acids long. In some embodiments, the tumor antigen peptides is about 29 amino acids long. In some embodiments, the tumor antigen peptides is about 30 amino acids long. In some embodiments, the tumor antigen peptides is about 31 amino acids long. In some embodiments, the tumor antigen peptides is about 32 amino acids long. In some embodiments, the tumor antigen peptides is about 33 amino acids long. In some embodiments, the tumor antigen peptides is about 34 amino acids long. In some embodiments, the tumor antigen peptides is about 35 amino acids long. In some embodiments, the tumor antigen peptides is about 36 amino acids long. In some embodiments, the tumor antigen peptides is about 37 amino acids long. In some embodiments, the tumor antigen peptides is about 38 amino acids long. In some embodiments, the tumor antigen peptides is about 39 amino acids long. In some embodiments, the tumor antigen peptides is about 40 amino acids long. In some embodiments, the tumor antigen peptides is about 41 amino acids long. In some embodiments, the tumor antigen peptides is about 42 amino acids long. In some embodiments, the tumor antigen peptides is about 43 amino acids long. In some embodiments, the tumor antigen peptides is about 44 amino acids long. In some embodiments, the tumor antigen peptides is about 45 amino acids long. In some embodiments, the tumor antigen peptides is about 46 amino acids long. In some embodiments, the tumor antigen peptides is about 47 amino acids long. In some embodiments, the tumor antigen peptides is about 48 amino acids long. In some embodiments, the tumor antigen peptides is about 49 amino acids long. In some embodiments, the tumor antigen peptides is about 50 amino acids long. In some embodiments, the tumor antigen peptides is about 51 amino acids long. In some embodiments, the tumor antigen peptides is about 52 amino acids long. In some embodiments, the tumor antigen peptides is about 53 amino acids long. In some embodiments, the tumor antigen peptides is about 54 amino acids long. In some embodiments, the tumor antigen peptides is about 55 amino acids long. In some embodiments, the tumor antigen peptides is about 56 amino acids long. In some embodiments, the tumor antigen peptides is about 57 amino acids long. In some embodiments, the tumor antigen peptides is about 58 amino acids long. In some embodiments, the tumor antigen peptides is about 59 amino acids long. In some embodiments, the tumor antigen peptides is about 60 amino acids long.

[0068] 60. In some embodiments, the tumor antigen peptides comprise at least one peptide comprising an MHC-I epitope or MHC-II epitope. 61. In some embodiments, disclosed herein, the population of antigen specific immune cells comprises T-helper cells (Thl), cytotoxic T cells (CTLs), or natural killer cells (NK).

[0069] 62. In some embodiments, disclosed herein, the population of antigen-presenting cells comprises one or more dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl), B cell, or macrophage. In some embodiments, disclosed herein, the population of antigen-presenting cells of any preceding aspects are activated, matured, or stimulated in the presence of a maturation culture. In some embodiments, disclosed herein the maturation culture comprises interleukin- ip (IL-i ), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), and polyinosinic:polycytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6). In some embodiments, disclosed herein the maturation culture for aDCl cells comprises interleukin- 13 (IL-ip), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), and polyinosinic:poly cytidylic acid (poly-I:C).

[0070] 63. Also, disclosed herein, in some aspects, the inhibitors of transforming growth factor beta (TGF-P) signaling are added to pre-maturation culture or the maturation culture of the ex vivo generated antigen-presenting cells. In some embodiments, the inhibitors of TGF-P signaling comprise LY364947, SD208, SB431542, an alternative small molecule, protein or RNA blocker of the TGF-P signaling or TGF-P production.

[0071] 64. In some embodiments, the population of antigen-presenting cells of any preceding aspect, produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response to CD40 ligand (CD40L) stimulation as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-P signaling. In some embodiments, aDCl produces at least 50% or more high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response after 24 hours of stimulation to CD40L as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-P signaling. In some embodiments, the ex vivo-generated antigen-presenting cell produces at least 50% or more high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response after 24 hours of stimulation to CD40L as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-P signaling.

[0072] 65. In one aspect, disclosed herein is a method of treating, inhibiting, reducing, decreasing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, melanoma, hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma, or brain cancer) in a subject comprising obtaining a population of peripheral blood mononuclear cells (PBMCs) from the subject; obtaining a population of antigen-presenting cells and a population of immune cells from the population of PBMCs; inducing maturation of the population of antigen-presenting cells, thereby obtaining mature antigen-presenting cells; blocking endogenous TGF- signaling in the mature antigen-presenting cells, thereby obtaining TGF-P inhibited antigen-presenting cells; contacting the TGF-P inhibited antigen-presenting cells with at least one or more tumor antigen peptides to obtain antigen loaded TGF-P inhibited antigen-presenting cells; co-culturing the antigen loaded TGF-P inhibited antigen-presenting cells with the population of immune cells to obtain the population of activated antigen specific immune cells; and administering to the subject an effective amount of the population of activated antigen specific immune cells.

[0073] 66. In some embodiments, the co-culturing period for the antigen loaded TGF-P inhibited antigen-presenting cells with the population of immune cells to obtain the population of activated antigen specific immune cells can be about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 hours. In some embodiments, the contacting period required for activation of immune cells can be about 1 hour. In some embodiments, the contacting period required for activation of immune cells can be about 2 hours. In some embodiments, the contacting period required for activation of immune cells can be about 3 hours. In some embodiments, the contacting period required for activation of immune cells can be about 4 hours. In some embodiments, the contacting period required for activation of immune cells can be about 5 hours. In some embodiments, the contacting period required for activation of immune cells can be about 6 hours. In some embodiments, the contacting period required for activation of immune cells can be about 7 hours. In some embodiments, the contacting period required for activation of immune cells can be about 8 hours. In some embodiments, the contacting period required for activation of immune cells can be about 9 hours. In some embodiments, the contacting period required for activation of immune cells can be about 10 hours. In some embodiments, the contacting period required for activation of immune cells can be about 11 hours. In some embodiments, the contacting period required for activation of immune cells can be about 12 hours. In some embodiments, the contacting period required for activation of immune cells can be about 13 hours. In some embodiments, the contacting period required for activation of immune cells can be about 14 hours. In some embodiments, the contacting period required for activation of immune cells can be about 15 hours. In some embodiments, the contacting period required for activation of immune cells can be about 16 hours. In some embodiments, the contacting period required for activation of immune cells can be about 17 hours. In some embodiments, the contacting period required for activation of immune cells can be about 18 hours. In some embodiments, the contacting period required for activation of immune cells can be about 19 hours. In some embodiments, the contacting period required for activation of immune cells can be about 20 hours.

[0074] 67. In some embodiments, disclosed herein the population of peripheral blood mononuclear cells (PBMCs) is autologous or allogeneic.

[0075] 68. In some embodiments, disclosed herein the antigen loaded TGF-P inhibited antigen- presenting cells comprises co-culturing with the population of immune cells for about 4 days to about 21 days.

[0076] 69. In some embodiments, disclosed herein the population of activated antigen specific immune cells are administered to the subject at least three times. In some embodiments, disclosed herein the interval between each administration of the population of activated antigen specific immune cells is about 0.5 month to about 2 months. In some embodiments, disclosed herein the population of activated antigen specific immune cells are administered by intradermal, intranodal, intravenous, intratumorally or peritumoral administration.

[0077] 70. In one aspect, disclosed herein is an ex vzvo-generated antigen-presenting cell, comprising culturing ex vzvo-generated antigen-presenting cell in the presence of inhibitors of transforming growth factor beta (TGF-P) signaling; and adding the inhibitors of TGF-P signaling to a pre-maturation culture or a maturation culture.

[0078] 71. In some embodiments, disclosed herein, the inhibitors of TGF-P signaling comprise LY364947, SD208, or SB431542. In some embodiments, disclosed herein the inhibitors of TGF- P signaling comprise an alternative small molecule, protein or RNA blocker of the TGF-P signaling or TGF-P production. In some embodiments, disclosed herein the ex vzvo-generated antigen-presenting cell comprises dendritic cell (DC). In some embodiments, disclosed herein the ex vzvo-generated antigen-presenting cell comprises alpha-type- 1 polarized dendritic cell (aDCl) exposed ex vivo to a combination of type-I and type-II interferons (IFNs); wherein the type-I IFNs comprise interferon-a (IFNa) or interferon-P (IFNP) and type-II IFNs comprise interferon-y (IFNy). In some embodiments, disclosed herein the interferon a (IFNa) is at a concentration of 1-10,000 U / ml. In some embodiments, disclosed herein the interferon y (IFNy) is at a concentration of 1-10,000 U / ml.

[0079] 72. In some embodiments, disclosed herein the aDCl is activated in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL- 1 P), tumor necrosis factor-a (TNFa), IFNa, IFNy, and a TLR3 ligand. In some embodiments, disclosed herein the TLR3 ligand is polyinosinic:polycytidylic acid (poly-I:C). In some embodiments, disclosed herein the IL-ip is at a concentration of 0.2-200 ng / ml. In some embodiments, disclosed herein the the TNFa is at a concentration of 0.2-200 ng / ml. In some embodiments, disclosed herein the IFNa is at a concentration of 1-10,000 U / ml. In some embodiments, disclosed herein the IFNy is at a concentration of 1- 10,000 U / ml.

[0080] 73. In some embodiments, disclosed herein the poly-I:C is at a concentration of 0.2 ng / ml-500 pg / ml. In some embodiments, the Poly inosinic:polycy tidy lie acid (poly-I:C) is at a concentration of 0.2 ng / ml-200 pg / ml.

[0081] 74. In some embodiments, disclosed herein the ex vivo -generated antigen-presenting cell produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 9 (CXCL9), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response to CD40 ligand (CD40L) stimulation as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-|3 signaling. In some embodiments, disclosed herein the said ex vzvo-generated antigen-presenting cell produces at least 50% or more of IL-12p70, IL-18, CXCL9, CXCL10, CXCL11, CCL5, or CCL19 in response after 24 hours of stimulation to CD40L as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-0 signaling.

[0082] 75. It is understood and herein contemplated that the disclosed treatment regimens can used alone or in combination with any anti-cancer therapy known in the art including, but not limited to Abemaciclib, Abiraterone Acetate, ABITREXATE® (Methotrexate), ABRAXANE® (Paclitaxel Albumin-stabilized Nanoparticle Formulation), ABVD, ABVE, ABVE-PC, AC, AC- T, ADCETRIS® (Brentuximab Vedotin), ADE, Ado-Trastuzumab Emtansine, ADRIAMYCIN® (Doxorubicin Hydrochloride), Afatinib Dimaleate, AFINITOR® (Everolimus), AKYNZEO® (Netupitant and Palonosetron Hydrochloride), ALDARA® (Imiquimod), Aldesleukin, ALECENSA® (Alectinib), Alectinib, Alemtuzumab, ALIMTA® (Pemetrexed Disodium), ALIQOPA® (Copanlisib Hydrochloride), ALKERAN™ for Injection (Melphalan Hydrochloride), ALKERAN™ Tablets (Melphalan), ALOXI® (Palonosetron Hydrochloride), ALUNBRIG® (Brigatinib), AMBOCHLORIN® (Chlorambucil), AMBOCLORIN® (Chlorambucil), Amifostine, Aminolevulinic Acid, Anastrozole, Aprepitant, AREDIA® (Pamidronate Disodium), ARIMIDEX® (Anastrozole), AROMASIN® (Exemestane),ARRANON® (Nelarabine), Arsenic Trioxide, ARZERRA® (Ofatumumab), Asparaginase Erwinia chrysanthemi, Atezolizumab, AVASTIN® (Bevacizumab), Avelumab, Axitinib, Azacitidine, BAVENCIO® (Avelumab), BEACOPP, BECENUM® (Carmustine), BELEODAQ® (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, BESPONSA® (Inotuzumab Ozogamicin) , Bevacizumab, Bexarotene, BEXXAR® (Tositumomab and Iodine I 131 Tositumomab), Bicalutamide, BICNU® (Carmustine), Bleomycin, Blinatumomab, BLINCYTO® (Blinatumomab), Bortezomib, BOSULIF® (Bosutinib), Bosutinib, Brentuximab Vedotin, Brigatinib, BuMel, Busulfan, BUSULFEX® (Busulfan), Cabazitaxel, CABOMETYX® (Cabozantinib-S -Malate), Cabozantinib-S-Malate, CAF, CAMPATH® (Alemtuzumab), CAMPTOSAR® (Irinotecan Hydrochloride), Capecitabine, CAPOX, CARAC® (Fluorouracil- Topical), Carboplatin, CARBOPLATIN-TAXOL, Carfilzomib, CARMUBRIS® (Carmustine), Carmustine, Carmustine Implant, CASODEX® (Bicalutamide), CEM, Ceritinib, CERUBIDINE® (Daunorubicin Hydrochloride), CERVARIX® (Recombinant HPV Bivalent Vaccine), Cetuximab, CEV, Chlorambucil, CHLORAMBUCIL-PREDNISONE, CHOP, Cisplatin, Cladribine, CLAFEN® (Cyclophosphamide), Clofarabine, CLOFAREX® (Cl of arabine), CLOLAR® (Clofarabine), CMF, Cobimetinib, COMETRIQ® (Cabozantinib-S- Malate), Copanlisib Hydrochloride, COPDAC, COPP, COPP-ABV, COSMEGEN® (Dactinomycin), COTELLIC® (Cobimetinib), Crizotinib, CVP, Cyclophosphamide, CYFOS® (Ifosfamide), CYRAMZA® (Ramucirumab), Cytarabine, Cytarabine Liposome, CYTOSAR- U® (Cytarabine), CYTOXAN® (Cyclophosphamide), Dabrafenib, Dacarbazine, DACOGEN® (Decitabine), Dactinomycin, Daratumumab, DARZALEX® (Daratumumab), Dasatinib, Daunorubicin Hydrochloride, Daunorubicin Hydrochloride and Cytarabine Liposome, Decitabine, Defibrotide Sodium, DEFITELIO® (Defibrotide Sodium), Degarelix, Denileukin Diftitox, Denosumab, DEPOCYT® (Cytarabine Liposome), Dexamethasone, Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, DOXIL® (Doxorubicin Hydrochloride Liposome), Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, DOX-SL® (Doxorubicin Hydrochloride Liposome), DTIC-DOME® (Dacarbazine), Durvalumab, EFUDEX® (Fluorouracil— Topical), ELITEK® (Rasburicase), ELLENCE® (Epirubicin Hydrochloride), Elotuzumab, ELOXATIN® (Oxaliplatin), Eltrombopag Olamine, EMEND® (Aprepitant), EMPLICITI® (Elotuzumab), Enasidenib Mesylate, Enzalutamide, Epirubicin Hydrochloride , EPOCH, ERBITUX® (Cetuximab), Eribulin Mesylate, ERIVEDGE® (Vismodegib), Erlotinib Hydrochloride, ERWINAZE® (Asparaginase Erwinia chrysanthemi), ETHYOL® (Amifostine), Etopophos ETOPOPHOS® (Etoposide Phosphate), Etoposide, Etoposide Phosphate, EVACET® (Doxorubicin Hydrochloride Liposome), Everolimus, EVISTA® (Raloxifene Hydrochloride), EVOMELA® (Melphalan Hydrochloride), Exemestane, 5-FU® (Fluorouracil Injection), 5-FU® (Fluorouracil— Topical), FARESTON® (Toremifene), FARYDAK® (Panobinostat), FASLODEX® (Fulvestrant), FEC, FEMARA® (Letrozole), Filgrastim, FLUDARA® (Fludarabine Phosphate), Fludarabine Phosphate, FLUOROPLEX® (Fluorouracil-Topical), Fluorouracil Injection, Fluorouracil— Topical, Flutamide, FOLEX® (Methotrexate), FOLEX PFS® (Methotrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, FOLOTYN® (Pralatrexate), FU-LV, Fulvestrant, GARDASIL® (Recombinant HPV Quadrivalent Vaccine), GARDASIL 9® (Recombinant HPV Nonavalent Vaccine), GAZYVA® (Obinutuzumab), Gefitinib, Gemcitabine Hydrochloride, GEMCITABINE-CISPLATIN, GEMCITABINE-OXALIPLATIN, Gemtuzumab Ozogamicin, GEMZAR® (Gemcitabine Hydrochloride), GILOTRIF® (Afatinib Dimaleate), GLEEVEC® (Imatinib Mesylate), GLIADEL® (Carmustine Implant), GLIADEL WAFER® (Carmustine Implant), Glucarpidase, Goserelin Acetate, HALAVEN® (Eribulin Mesylate), HEMANGEOL® (Propranolol Hydrochloride), HERCEPTIN® (Trastuzumab), HPV Bivalent Vaccine, Recombinant, HPV Nonavalent Vaccine, Recombinant, HPV Quadrivalent Vaccine, Recombinant, HYCAMTIN® (Topotecan Hydrochloride), HYDREA® (Hydroxyurea), Hydroxyurea, Hyper-CVAD, IBRANCE® (Palbociclib), Ibritumomab Tiuxetan, Ibrutinib, ICE, ICLUSIG® (Ponatinib Hydrochloride), IDAMYCIN® (Idarubicin Hydrochloride), Idarubicin Hydrochloride, Idelalisib, IDHIFA® (Enasidenib Mesylate), IFEX® (Ifosfamide), Ifosfamide, IFOSFAMIDUM® (Ifosfamide), IL-2 (Aldesleukin), Imatinib Mesylate, IMBRUVICA® (Ibrutinib), IMFINZI® (Durvalumab), Imiquimod, IMLYGIC® (Talimogene Laherparepvec), INLYTA® (Axitinib), Inotuzumab Ozogamicin, Interferon Alfa-2b, Recombinant, Interleukin-2 (Aldesleukin), INTRON A® (Recombinant Interferon Alfa-2b), Iodine I 131 Tositumomab and Tositumomab, Ipilimumab, IRESSA® (Gefitinib), Irinotecan Hydrochloride, Irinotecan Hydrochloride Liposome, ISTODAX® (Romidepsin), Ixabepilone, Ixazomib Citrate, IXEMPRA® (Ixabepilone), JAKAFI® (Ruxolitinib Phosphate), JEB, JEVTANA® (Cabazitaxel), KADCYLA® (Ado-Trastuzumab Emtansine), KEOXIFENE® (Raloxifene Hydrochloride), KEPIVANCE® (Palifermin), KEYTRUDA® (Pembrolizumab), KISQALI® (Ribociclib), KYMRIAH® (Tisagenlecleucel), KYPROLIS® (Carfilzomib), Lanreotide Acetate, Lapatinib Ditosylate, LARTRUVO® (Olaratumab), Lenalidomide, Lenvatinib Mesylate, LENVIMA® (Lenvatinib Mesylate), Letrozole, Leucovorin Calcium, LEUKERAN® (Chlorambucil), Leuprolide Acetate, LEUSTATIN® (Cladribine), LEVULAN® (Aminolevulinic Acid), LINFOLIZIN® (Chlorambucil), LIPODOX® (Doxorubicin Hydrochloride Liposome), Lomustine, LONSURF® (Trifluridine and Tipiracil Hydrochloride), LUPRON® (Leuprolide Acetate), LUPRON DEPOT® (Leuprolide Acetate), LUPRON DEPOT-PED® (Leuprolide Acetate), LYNPARZA® (Olaparib), MARQIBO® (Vincristine Sulfate Liposome), MATULANE® (Procarbazine Hydrochloride), Mechlorethamine Hydrochloride, Megestrol Acetate, MEKINIST® (Trametinib), Melphalan, Melphalan Hydrochloride, Mercaptopurine, Mesna, MESNEX® (Mesna), METHAZOLASTONE® (Temozolomide), Methotrexate, METHOTREXATE LPF® (Methotrexate), Methylnaltrexone Bromide, MEXATE® (Methotrexate), MEXATE-AQ® (Methotrexate), Midostaurin, Mitomycin C, Mitoxantrone Hydrochloride, MTTOZYTREX® (Mitomycin C), MOPP, MOZOBIL® (Plerixafor), MUSTARGEN® (Mechlorethamine Hydrochloride) , MUTAMYCIN® (Mitomycin C), MYLERAN® (Busulfan), MYLOSAR® (Azacitidine), MYLOTARG® (Gemtuzumab Ozogamicin), NANOPARTICLE PACLITAXEL® (Paclitaxel Albumin- stabilized Nanoparticle Formulation), NAVELBINE® (Vinorelbine Tartrate), Necitumumab, Nelarabine, NEOSAR® (Cyclophosphamide), Neratinib Maleate, NERLYNX® (Neratinib Maleate), Netupitant and Palonosetron Hydrochloride, NEULASTA® (Pegfilgrastim), NEUPOGEN® (Filgrastim), NEXAVAR® (Sorafenib Tosylate), NILANDRON® (Nilutamide), Nilotinib, Nilutamide, NINLARO® (Ixazomib Citrate), Niraparib Tosylate Monohydrate, Nivolumab, NOLVADEX® (Tamoxifen Citrate), NPLATE® (Romiplostim), Obinutuzumab, ODOMZO® (Sonidegib), OEPA, Ofatumumab, OFF, Olaparib, Olaratumab, Omacetaxine Mepesuccinate, ONCASPAR® (Pegaspargase), Ondansetron Hydrochloride, ONIVYDE® (Irinotecan Hydrochloride Liposome), ONTAK® (Denileukin Diftitox), OPDIVO® (Nivolumab), OPPA, Osimertinib, Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized Nanoparticle Formulation, PAD, Palbociclib, Palifermin, Palonosetron Hydrochloride, Palonosetron Hydrochloride and Netupitant, Pamidronate Disodium, Panitumumab, Panobinostat, PARAPLAT® (Carboplatin), PARAPLATIN® (Carboplatin), Pazopanib Hydrochloride, PCV, PEB, Pegaspargase, Pegfilgrastim, Peginterferon Alfa- 2b, PEGINTRON® (Peginterferon Alfa-2b), Pembrolizumab, Pemetrexed Disodium, PERJETA® (Pertuzumab), Pertuzumab, PLATINOL® (Cisplatin), PLATINOL-AQ® (Cisplatin), Plerixafor, Pomalidomide, POMALYST® (Pomalidomide), Ponatinib Hydrochloride, PORTRAZZA® (Necitumumab), Pralatrexate, Prednisone, Procarbazine Hydrochloride, PROLEUKIN® (Aldesleukin), PROLIA® (Denosumab), PROMACTA® (Eltrombopag Olamine), Propranolol Hydrochloride, PROVENGE® (Sipuleucel-T), PURINETHOL® (Mercaptopurine), PURIXAN® (Mercaptopurine), Radium 223 Dichloride, Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R-CVP, Recombinant Human Papillomavirus (HPV) Bivalent Vaccine, Recombinant Human Papillomavirus (HPV) Nonavalent Vaccine, Recombinant Human Papillomavirus (HPV) Quadrivalent Vaccine, Recombinant Interferon Alfa-2b, Regorafenib, RELISTOR® (Methylnaltrexone Bromide), R-EPOCH, REVLIMID® (Lenalidomide), RHEUMATREX® (Methotrexate), Ribociclib, R-ICE, R1TUXAN® (Rituximab), RITUXAN HYCELA® (Rituximab and Hyaluronidase Human), Rituximab, Rituximab and , Hyaluronidase Human, ,Rolapitant Hydrochloride, Romidepsin, Romiplostim, RUBIDOMYCIN® (Daunorubicin Hydrochloride), RUBRACA® (Rucaparib Camsylate), Rucaparib Camsylate, Ruxolitinib Phosphate, R YD APT® (Midostaurin), Sclerosol Intrapleural Aerosol (Talc), Siltuximab, Sipuleucel-T, SOMATULTNE DEPOT® (Lanreotide Acetate), Sonidegib, Sorafenib Tosylate, SPRYCEL® (Dasatinib), STANFORD V, Sterile Talc Powder (Talc), STERITALC® (Talc), STIVARGA® (Regorafenib), Sunitinib Malate, SUTENT® (Sunitinib Malate), SYLATRON® (Peginterferon Alfa- 2b), SYLVANT® (Siltuximab), Synribo SYNRIBO® (Omacetaxine Mepesuccinate), TABLOID® (Thioguanine), TAC, TAFINLAR® (Dabrafenib), TAGRISSO® (Osimertinib), Talc, Talimogene Laherparepvec, Tamoxifen Citrate, TARABINE PFS® (Cytarabine), TARCEVA® (Erlotinib Hydrochloride), TARGRETIN® (Bexarotene), TASIGNA® (Nilotinib), TAXOL® (Paclitaxel), TAXOTERE® (Docetaxel), TECENTRIQ® (Atezolizumab), TEMODAR® (Temozolomide), Temozolomide, Temsirolimus, Thalidomide, THALOMID® (Thalidomide), Thioguanine, Thiotepa, Tisagenlecleucel, TOLAK® (Fluorouracil— Topical), Topotecan Hydrochloride, Toremifene, TORISEL® (Temsirolimus), Tositumomab and Iodine I 131 Tositumomab, TOTECT® (Dexrazoxane Hydrochloride), TPF, Trabectedin, Trametinib, Trastuzumab, TREANDA® (Bendamustine Hydrochloride), Trifluridine and Tipiracil Hydrochloride, TRISENOX® (Arsenic Trioxide), TYKERB® (Lapatinib Ditosylate) , UNITUXIN® (Dinutuximab), Uridine Triacetate, VAC, Vandetanib, VAMP, VARUBI® (Rolapitant Hydrochloride), VECTIBIX® (Panitumumab), VelP, VELBAN® (Vinblastine Sulfate), VELCADE® (Bortezomib), VELSAR® (Vinblastine Sulfate), Vemurafenib, VENCLEXTA® (Venetoclax), Venetoclax, VERZENIO® (Abemaciclib), VIADUR® (Leuprolide Acetate), VIDAZA® (Azacitidine), Vinblastine Sulfate, VINCASAR PFS® (Vincristine Sulfate), Vincristine Sulfate, Vincristine Sulfate Liposome, Vinorelbine Tartrate, VIP, Vismodegib, VISTOGARD® (Uridine Triacetate), VORAXAZE® (Glucarpidase), Vorinostat, VOTRIENT® (Pazopanib Hydrochloride), VYXEOS® (Daunorubicin Hydrochloride and Cytarabine Liposome), WELLCOVORIN® (Leucovorin Calcium), XALKORI® (Crizotinib), XELODA® (Capecitabine), XELIRI, XELOX, XGEVA® (Denosumab), XOFIGO® (Radium 223 Dichloride), XT ANDI® (Enzalutamide), YERVOY® (Ipilimumab), YONDELIS® (Trabectedin), ZALTRAP® (Ziv-Aflibercept), ZARXIO® (Filgrastim), ZEJULA® (Niraparib Tosylate Monohydrate), ZELBORAF® (Vemurafenib), ZEVALIN® (Ibritumomab Tiuxetan), ZINECARD® (Dexrazoxane Hydrochloride), Ziv- Aflibercept, ZOFRAN® (Ondansetron Hydrochloride), ZOLADEX® (Goserelin Acetate), Zoledronic Acid, ZOLINZA® (Vorinostat), ZOMETA® (Zoledronic Acid), ZYDELIG® (Idelalisib), ZYKADIA® (Ceritinib), and / or ZYTIGA® (Abiraterone Acetate). The treatment methods can include or further include checkpoint inhibitors including, but are not limited to antibodies that block PD-1 (such as, for example, Nivolumab (BMS-936558 or MDX1106), pembrolizumab, cemiplimab , CT-011, MK-3475), PD-L1 (such as, for example, atezolizumab, avelumab, durvalumab, MDX-1 105 (BMS-936559), MPDL3280A, or MSB0010718C), PD-L2 (such as, for example, rHIgM12B7), CTLA-4 (such as, for example, Ipilimumab (MDX-010), Tremelimumab (CP-675,206)), IDO, B7-H3 (such as, for example, MGA271, MGD009, omburtamab), B7-H4, B7-H3, T cell immunoreceptor with Ig and HIM domains (TIGIT)(such as, for example BMS-986207, OMP-313M32, MK-7684, AB-154, ASP-8374, MTIG7192A, or PVSRIPO), CD96, B- and T-lymphocyte attenuator (BTLA), V-domain Ig suppressor of T cell activation (VISTA)(such as, for example, JNJ-61610588, CA-170), TIM3 (such as, for example, TSR-022, MBG453, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, RO7121661), LAG-3 (such as, for example, BMS-986016, LAG525, MK-4280, REGN3767, TSR-033, BI754111, Sym022, FS118, MGD013, and Immutep).

[0083] DC-based cancer vaccines

[0084] 76. DC-based vaccines have been increasingly applied in the clinical treatment of patients with cancer. Following the initial success of the multi-epitope melanoma trial (30% objective clinical responses), DCs have been used successfully to treat patients with melanoma, lymphoma, and renal cell carcinoma, with long-term responses observed in many of them. However, overall clinical response rates and the survival benefit of the early generation DC vaccines were disappointing, highlighting the need to improve the design of DC-based vaccines, notably the selection of the most appropriate types of DCs and vaccination regimen. Fully-mature DCs induced by IL-ip, TNFcx, IL-6, and PGET have been consistently observed as superior to immature DCs, in their ability to migrate to the vaccine-draining lymph nodes (LN) and to induce higher numbers of cancer-specific specific T cells in vivo. DCs induced by this “complete cytokine mix” are generally considered as the current “gold standard” in DC-based cancer immunotherapy. However, despite their mature status and their ability to induce IFNy- producing T cells in vivo, the therapeutic efficacy of these cells was disappointing. One factor restricting their efficacy can be their relatively low production of IL-12p70. Unfortunately, the PGE2-based maturation cocktail reduces the ability of DCs to produce IL-12p70 and supports the preferential secretion of IL-12-antagonistic p40 homodimer. For this reason, the desirable combination of high immunostimulatory activity and high responsiveness to lymph-node- directing chemokines, with high capacity to produce IL-12p70 has not been attained by DC preparations used in the past clinical trials.

[0085] 77. In one aspect, disclosed herein are cancer vaccines comprising a population of antigen-presenting cells (including, but not limited to dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage) loaded ex vivo with at least one or more tumor antigen peptides. In some aspects, the disclosed cancer vaccine can be used for treating, inhibiting, reducing, decreasing, ameliorating, and / or preventing a cancer and / or metastasis (such as, for example, glioma, brain cancer (including, but not limited to recurrent brain cancer), anaplastic astrocytoma (AA), hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma and melanoma (including, but not limited to checkpoint-resistant metastatic melanoma)) in a subject comprising a population of antigen-presenting cells loaded ex vivo with at least one or more tumor antigen peptides, wherein the population of antigen-presenting cells comprises dendritic cell (DC) or alpha-type- 1 polarized dendritic cell (aDCl) with inhibited endogenous transforming growth factor beta (TGF-P) signaling.

[0086] 78. In some embodiments, disclosed herein, the dendritic cell (DC) or alpha-type- 1 polarized dendritic cell (aDCl) with inhibited endogenous TGF-P signaling are activated, matured, or stimulated in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL-ip), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), and polyinosinic:poly cytidylic acid (poly-I:C).

[0087] 79. In some embodiments, disclosed herein, the cancer vaccine, further comprises addition of TGF-P signaling inhibitors to pre-maturation culture or the maturation culture, wherein the inhibitors of TGF-P signaling comprise LY364947, SD208 or SB431542.

[0088] 80. In some embodiments, disclosed herein, the cancer vaccine produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCLl 1), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

[0089] 81. In some embodiments, disclosed herein, the population of antigen-presenting cells of any preceding aspect, loaded ex vivo with at least one or more tumor antigen peptides are autologous or allogeneic.

[0090] 82. In one aspect, disclosed herein is a cancer vaccine comprising an ex vivo-generated antigen-presenting cell of any preceding aspect, loaded ex vivo with at least one or more tumor antigen peptides. 83. In one aspect, disclosed herein is an activated antigen specific T-cell, comprising activating the antigen specific T-cell with an ex vi vo-generated antigen-presenting cell of any preceding aspect.

[0091] 84. In some embodiments, disclosed herein, the activated antigen specific T-cell is administered intratumorally.

[0092] 85. In one aspect disclosed herein is an ex vzvo-generated antigen-presenting cell, obtained by the method comprising culturing ex vzvo-generated antigen-presenting cell (including, but not limited to dendritic cell (DC), B cell, or macrophage) in the presence of inhibitors of transforming growth factor beta (TGF-P) signaling (such as, for example, small molecule, protein or RNA blocker of the TGF-P signaling or TGF- production including, but not limited to LY364947, SD208 or SB431542); and adding the inhibitors of TGF-P signaling to a pre-maturation culture or a maturation culture. In some aspects, the ex vzvo-generated antigen- presenting cell comprises alpha- type- 1 polarized dendritic cell (aDCl) exposed ex vivo to a combination of type-I and type-II interferons (IFNs); wherein the type-I IFNs comprise interferon-a (IFNa) or interferon-P (IFNP) and type-II IFNs comprise interferon-y (IFNy). In some aspects, the aDCl is activated in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL-ip), tumor necrosis factor-a (TNFa), IFNa, IFNy, and a TLR3 ligand (such as, for example, polyinosinic:poly cytidylic acid (poly-I:C)

[0093] 86. Also disclosed herein is an ex vzvo-generated antigen-presenting cell, wherein the Interleukin ip (IL1P) is at a concentration of about 0.2-200 ng / ml, the Tumor necrosis factor a (TNFa) is at a concentration of about 0.2-200 ng / ml, the Interferon a (IFNa) is at a concentration of about 1-10,000 U / ml, the Interferon y (IFNy) is at a concentration of about 1-10,000 U / ml and / or the TLR3 ligand (such as for example, Polyinosinic:polycytidylic acid (poly-I:C)) is at a concentration of 0.2 ng / ml - 500 pg / ml.

[0094] 87. In one aspect, disclosed herein is an ex vzvo-generated antigen-presenting cell, wherein the said ex vzvo-generated antigen-presenting cell produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 9 (CXCL9, )C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response to CD40 ligand (CD40L) stimulation as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-P signaling.

[0095] 88. Also disclosed herein is an ex vzvo-generated antigen-presenting cell, wherein the said ex vzvo-generated antigen-presenting cell produces at least 50% or more of IL-12p70, IL-18, CXCL9, CXCL10, CXCL11, CCL5, or CCL19 in response after 24 hours of stimulation to CD40L as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-0 signaling.

[0096] 89. In one aspect disclosed herein is a cancer vaccine comprising any of the ex vivo- generated antigen-presenting cells disclosed herein loaded ex vivo with at least one or more tumor antigen peptides.

[0097] 90. Also disclosed herein are activated antigen specific T-cells, wherein the T cell is activated with the any of the ex v / vo-generated antigen-presenting cells disclosed herein.

[0098] VII. EXAMPLES

[0099] Example 1: Type-1 polarized DC (DC1) are superior inducers of Thl cells, CTLs, and cytotoxic NK cells

[0100] 91. It was observed that DCs matured in the presence of IFN-y (type-1 polarized DC or DC1) produce up to 100-fold higher levels of IL-12p70 in response to CD40L stimulation, than similarly mature DCs induced by LPS alone or by the combination of IL- 1 [3 and TNFa. Importantly, DCls are resistant to tumor-associated immunosuppressive factors, and can produce lL-12p70 upon the interaction with CD4+T cells (such as naive Th cells) that are themselves unable to produce IFNy. These features indicate that DCls can be also effective in immunocompromised cancer patients.

[0101] 92. It was observed that IFNa and poly-I:C, a synthetic analogue of dsRNA allows for the generation of DC1 in serum-free cultures (aDCls), paving the way for the clinical application of cancer vaccines based on the paradigm of DC polarization. When compared to the current “gold standard” DCs (matured by IL- 10, TNFa, IL-6, and PGE2), aDCls induce up to 40-fold more CTLs specific for MART-1, gplOO, and tyrosinase, after a single round of in vitro sensitization. They also strongly enhance cytotoxic Thl -type CD4+T cell responses, and the tumoricidal activity of NK cells. Therefore, while the currently used DC-based vaccines rely on either immature DCs with high ability to produce IL- 12 but low stimulatory / LN-homing capacity, or mature DCs with high stimulatory / LN-homing functions, but reduced ability to produce IL-12, the possibility to generate DCls allows us for the first time to combine all of these desirable features within a single vaccine preparation.

[0102] Example 2: Alpha-type-l-polarized dendritic cells (aDCls): Clinical activity in patients with recurrent brain cancer and checkpoint-resistant metastatic melanoma

[0103] 93. The ability of aDCls, monocyte-derived DCs matured in the conditions mimicking virally induced acute inflammation (IL-10 / TNFa / IFNo / IFNy / poly-LC) to selectively induce and amplify CTLs, Thl and NK cell responses, led to their applications in the therapy of cancer. It is demonstrated that aDCl loaded with glioma- specific peptide epitopes (IL-13Ra2345-353:iA9v, gpl00209-2i7:2M, EphA2§83-89i and YKL-40201-210) combined with a TLR3 ligand, poly-ICLC, which induces the intratumoral production of CXCL9, CXCL10, CXCL11 and CCL5 which attract the CXCR3 and CCR5 expressing CTLs and Thl cells to multiple tumors, including brain tumors, can induce clinical responses and long-term disease stabilizations even in in patients with particularly aggressive recurrent primary brain cancers (high-grade gliomas: AA and GBM (FIG. 1A).

[0104] 94. Among the 19 vaccinated patients with recurrent high-grade gliomas (65% GBM, average progression-free survival: 2 months; 35% anaplastic astrocytoma; average progression- free survival: 4 months) they have obseved PFS of at least 12 months in 9 out of 19 patients. Even more strikingly 4 (21%) patients developed objective radiologic responses, and two patients (11%) survived at least 5 years. The demonstration that DC1 vaccines loaded with the tumor-relevant peptide antigens can induce long-term stabilizations and objective clinical responses in patients with brain cancer (recurrent GBM), traditionally considered an immunoprivileged site, provides strong evidence for the ability of aDCl vaccination in BMBC to overcome the immunosuppressive tumor microenvironment in the brain.

[0105] 95. The most recent trial in metastatic checkpoint-resistant melanoma (NCT01876212) utilizing aDCls loaded with six antigenic peptides targeting tumor-associated blood vessel antigens (TBVA; uniformly expressed on blood vessels of multiple human and mouse tumors has demonstrated massive epitope spreading and objective radiologic responses in 4 patients (and additional 2 patients with stable disease of at least 9 months), in 12 patients who had previously progressed on PD-1 blockade alone or combined PD-l / CTEA-4 blockade, including 4 objective responses in 7 patients who had previously shown primary PD-lresistance (FIG. 1C).

[0106] 96. These data demonstarte that aDCl loaded with antigens relevant to different tumors are able to induce effective immunity even against most agressive tumors, such as brain tumors or checkpoint-resistant meatastatic melanoma.

[0107] Example 3: Importance of selective induction and attraction of CTLs and Thl cells, but not Tregs

[0108] 97. CTL infiltration of the tumor predicts both improved response to therapy and overall survival. This is true in multiple other tumor types. It also and facilitates responsiveness of multiple cancer types to checkpoint blockers (such as PD-1 / PD-L1 / PD-L2 and CTLA-4, as well as TIM3 or LAG3) the new class of potent immunotherapeutic agents. In contrast, intratumoral prevalence of regulatory T cells (Tregs) is associated with poor tumor response and impaired survival in patients with many types of cancer. These findings highlight the ability for modulating TIL densities in the management of cancer, by selective induction of CTLs and Thl cells, rather than Tregs, or their selective attraction. Accordingly, enhanced infiltration of CD8+T cells following modulation of TME can improve spontaneous control of tumor growth and the effectiveness of PD-1 and CTLA-4 blockade in multiple mouse models. Importantly, for this area of cancer therapies, it is shown that “non-exhausted” DDCls selectively produce the chemokines which attract the desirable CTL (both memory and effector), Thl and NK cells, as well as other (CCL5, CCL19, CXCL10, CXCL11), as well as factors enhancing their functions, such as IL-12p70 or IL-18, but lower levels of “suppressor / regulatory” factors, such as CCL22, and preferentially attract CTLs, rather than Tregs. DC Is have also been shown to reprogram the Th2 cells into Thl cells and promote enhances attraction and activation of NK cells.

[0109] Example 4: Alternative use of aDCls as modulators of TME and in situ vaccines

[0110] 98. Mature ex vivo antigen-loaded DCs have been traditionally used as “vaccines” to induce tumor-specific T cells in draining lymph nodes. aDCls show enhanced ability to crosspresent tumor-derived antigens to CD8+T cells recognizing MHC class-I restricted tumor-related antigens in vitro and in vivo, and effectively convert non-cytolytic CD8+T cells into high perforin- and granzyme B -expressing CXCR3hlgh / CCR5hlghCTLs, with high killer activity, leading to their application as cancer vaccines after loading with antigens ex vivo.

[0111] 99. However, DCs are also essential in the effector phase of immune responses against cancer and other pathogens, with an especially high level of DC defect observed within the TME itself. In addition, multiple cancers do not have clearly defined tumor rejection antigens, limiting the applicability of antigenic peptides as the source of tumor-relevant antigens. The use of autologous cancer cells as the source of tumor-relevant antigens bypasses this problem (and allows treatment of the roughly 50% of cancer patients in US population who do not express HLA-A2), but autologous tumor- loaded DCs are logistically difficult to prepare and can only be administered 6 weeks or longer after tumor resection (production of the DC vaccine can only start after tumor resection and verification of successful isolation of cancer cells, then the generation and complete sterility testing takes at least 5 weeks). For this reason, they cannot be used at the moment when they are particularly needed, to prevent the early repopulation of the peritoneal cavity with residual cancer cells which evade chemotherapy and surgery. These concerns led to efforts to inject DCs intratumorally to take up the most relevant antigen, attract the desirable types of immune cells (CTLs, Thl and NK cells, and additional DCs), to convert cold-tumors into hot and, ideally convert the sites of injection into in-situ vaccines, further elevating the need for the production of the relevant chemokines, such as CXCL9, CXCL10, CXCL11, CCL5 and CCL19. 100. In contrast to the well-documented ability of exogenous TGF-P (TGF-P; known to be produced by tumors and healthy tissues), which has been known to limit the immunostimulatory functions of DCs, the DCs are not generally acknowledged as major source of TGF-P, and the ability for the enhancement of DCs functions by blocking endogenous TGF-P in DC cultures has not been studied. This makes the current observations novel and unexpected. It is shown that DCs matured in the presence of aDC 1 maturation cocktail and inhibitors of TGF-P signaling are qualitatively superior to the current aDC Is with regard to the production of IL- 12p70 and CTL / Th l / NK cell-attracting chemokines (FIG. 3). The data indicates that adding TGF-P blockers to the initial pre-maturation cultures and the maturation culture can enhance the DC production of both the IL-12p70 (FIGS. 3A, 3C and 3E) and the desirable chemokines (FIGS. 3B, 3D and 3F).

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Claims

IX. CLAIMSWhat is claimed is:

1. A method of treating cancer in a subject, comprising: activating a population of antigen specific immune cells in vitro by co-culturing with a population of antigen-presenting cells loaded ex vivo with at least one or more tumor antigen peptides, thereby obtaining activated antigen specific immune cells; and administering to the subject an effective amount of the activated antigen specific immune cells, wherein the population of antigen specific immune cells are activated prior to administration.

2. The method of claim 1 , wherein the population of antigen specific immune cells comprises co-culturing with the population of antigen-presenting cells loaded with at least one or more tumor antigen peptides for about 4 days to about 21 days.

3. The method of any one of claims 1-2, wherein the tumor antigen peptides is about 20 to about 40 amino acids long.

4. The method of any one of claims 1-3, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-I epitope.

5. The method of any one of claims 1-3, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-II epitope.

6. The method of any one of claims 1-5, wherein the cancer comprises melanoma, hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma, or brain cancer.

7. The method of any one of claims 1-6, wherein the population of antigen specific immune cells comprises T-helper cells (Th 1 ), cytotoxic T cells (CTLs), or natural killer cells (NK).

8. The method of any one of claims 1-7, wherein the population of antigen-presenting cells comprises one or more dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage.

9. The method of claims 1-8, wherein the one or more DC are matured in presence of a maturation culture, wherein the maturation culture comprises interleukin- 1 f> (IL- 10), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), polyinosinic:polycytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6).

10. The method of claims 1-9, further comprises addition of inhibitors of transforming growth factor beta (TGF-P) signaling to pre-maturation culture or the maturation culture, wherein the inhibitors of TGF-P signaling comprise LY364947, SD208 or SB431542,11. The method of claim 8, wherein aDCl produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL- 18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

12. A method of treating a cancer in a subject, comprising: administering to the subject in situ an effective amount of a population of antigen-presenting cells, loaded with at least one or more tumor antigen peptides.

13. The method of claim 12, wherein the tumor antigen peptides is about 20 to about 40 amino acids long.

14. The method of any one of claims 12-13, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-I epitope.

15. The method of any one of claims 12-13, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-II epitope.

16. The method of any one of claims 12-15, wherein the cancer comprises melanoma, hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma, or brain cancer.

17. The method of any one of claims 12-16, wherein the population of antigen-presenting cells comprises one or more dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl), B cell, or macrophage.

18. The method of claims 12-17, wherein the one or more DC are matured in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL- 1 ), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), polyinosinic:polycytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6).

19. The method of claims 12-18, further comprises addition of inhibitors of transforming growth factor beta (TGF- ) signaling to pre-maturation culture or the maturation culture.

20. The method of claims 12-19, wherein the inhibitors of TGF- signaling comprise LY364947, SD208 or SB431542.

21. The method of claim 17, wherein aDCl produces high levels of interleukin- 12 (IL- 12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

22. A cancer vaccine comprising a population of antigen-presenting cells loaded ex vivo with at least one or more tumor antigen peptides, wherein the population of antigen-presenting cells comprises dendritic cell (DC) or alpha-type- 1 polarized dendritic cell (aDCl) with inhibitedendogenous transforming growth factor beta (TGF-P) signaling23. The cancer vaccine of claim 22, wherein the dendritic cell (DC) or alpha-type-1 polarized dendritic cell (aDCl) with inhibited endogenous TGF-P signaling are matured in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL-ip), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), polyinosinic:polycytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6).

24. The cancer vaccine of claims 22-23, further comprises addition of TGF-P signaling inhibitors to pre-maturation culture or the maturation culture.

25. The cancer vaccine of claims 22-24, wherein the inhibitors of TGF- signaling comprise LY364947, SD208 or SB431542.

26. The cancer vaccine of claims 22-25, wherein the cancer vaccine produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL-18), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

27. The cancer vaccine of claims 22-26, wherein the tumor antigen peptides is about 20 to about 40 amino acids long.

28. The cancer vaccine of claims 22-27, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-I epitope.

29. The cancer vaccine of claims 22-27, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-II epitope.

30. The cancer vaccine of claims 22-29, wherein the cancer comprises melanoma, hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma, or brain cancer.

31. The cancer vaccine of any one of claims 22-30, wherein the population of antigen- presenting cells loaded ex vivo with at least one or more tumor antigen peptides are autologous.

32. The cancer vaccine of any one of claims 22-30, wherein population of antigen-presenting cells loaded ex vivo with at least one or more tumor antigen peptides are allogeneic.

33. A method of treating a cancer in a subject using a population of activated antigen specific immune cells, comprising:• obtaining a population of peripheral blood mononuclear cells (PBMCs) from the subject;• obtaining a population of antigen-presenting cells and a population of immune cells from the population of PBMCs;• inducing maturation of the population of antigen-presenting cells, thereby obtaining mature antigen-presenting cells;• blocking endogenous TGF- signaling in the mature antigen-presenting cells, thereby obtaining TGF-P inhibited antigen-presenting cells;• contacting the TGF-P inhibited antigen-presenting cells with at least one or more tumor antigen peptides to obtain antigen loaded TGF-P inhibited antigen- presenting cells;• co-culturing the antigen loaded TGF-P inhibited antigen-presenting cells with the population of immune cells to obtain the population of activated antigen specific immune cells; and• administering to the subject an effective amount of the population of activated antigen specific immune cells.

34. The method of claim 33, wherein the antigen loaded TGF-P inhibited antigen-presenting cells comprises co-culturing with the population of immune cells for about 4 days to about 21 days.

35. The method of any one of claims 33-34, wherein the tumor antigen peptides is about 20 to about 40 amino acids long.

36. The method of any one of claims 33-35, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-I epitope.

37. The method of any one of claims 33-35, wherein the tumor antigen peptides comprise at least one peptide comprising an MHC-II epitope.

38. The method of any one of claims 33-37, wherein the cancer comprises melanoma, hepatic cellular carcinoma, cervical cancer, lung cancer, colorectal cancer, lymphoma, renal carcinoma, breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, prostate cancer, nasopharyngeal carcinoma, or brain cancer.

39. The method of any one of claims 33-38, wherein the population of immune cells comprises T-helper cells (Thl), cytotoxic T cells (CTLs), or natural killer cells (NK).

40. The method of any one of claims 33-39, wherein the population of antigen-presenting cells comprises one or more dendritic cell (DC), alpha-type- 1 polarized dendritic cell (aDCl) B cell, or macrophage.

41. The method of claims 33-40, wherein the one or more DC are matured in presence of a maturation culture, wherein the maturation culture comprises interleukin- ip (IL- 1 ), tumor necrosis factor-a (TNFa), interferon-a (IFNa), interferon-y (IFNy), polyinosinic:polycytidylic acid (poly-I:C), prostaglandin E2 (PGE2), or interleukin-6 (IL-6).

42. The method of claims 33-41, further comprises addition of inhibitors of transforming growth factor beta (TGF-P) signaling to pre-maturation culture or the maturation culture.

43. The method of claims 33-42, wherein the inhibitors of TGF-P signaling comprise LY364947, SD208 or SB431542.

44. The method of claim 40, wherein aDCl produces high levels of interleukin- 12 (IL- 12p70), interleukin-18 (TL-18), C-X-C motif chemokin e ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19).

45. The method of claims 33-44, wherein the population of activated antigen specific immune cells are administered to the subject at least three times.

46. The method of claims 33-45, wherein the interval between each administration of the population of activated antigen specific immune cells is about 0.5 month to about 2 months.

47. The method of claims 33-46, wherein the population of activated antigen specific immune cells are administered by intradermal, intranodal, intravenous, intratumorally or peritumoral administration.

48. An ex vzvo-generated antigen-presenting cell, comprising: culturing ex vivo -generated antigen-presenting cell in the presence of inhibitors of transforming growth factor beta (TGF-P) signaling; and adding the inhibitors of TGF-P signaling to a pre-maturation culture or a maturation culture.

49. The ex row-generated antigen-presenting cell of claim 48, wherein the inhibitors of TGF- P signaling comprise LY364947, SD208, or SB431542.

50. The ex vz'vo-generated antigen-presenting cell of claims 48-49, wherein the inhibitors of TGF-P signaling comprise an alternative small molecule, protein or RNA blocker of the TGF-P signaling or TGF-P production.

51. The ex vzvo-generated antigen-presenting cell of claims 48-50, wherein the ex vivo- generated antigen-presenting cell comprises dendritic cell (DC).

52. The ex vi iw-generated antigen-presenting cell of claims 48-51, wherein the ex vivo- generated antigen-presenting cell comprises alpha-type-1 polarized dendritic cell (aDCl) exposed ex vivo to a combination of type-I and type-II interferons (IFNs); wherein the type-I IFNs comprise interferon-a (IFNa) or interferon-P (IFNP) and type-II IFNs comprise interferon- / (IFNy).

53. The ex vzvo-generated antigen-presenting cell of claims 48-52, wherein the interferon a (IFNa) is at a concentration of 1-10,000 U / ml.

54. The ex vzvc-generated antigen-presenting cell of claims 48-53, wherein the interferon y (IFNy) is at a concentration of 1-10,000 U / ml.

55. The ex vzi’o-generated antigen-presenting cell of claims 48-54, wherein the aDCl is activated in presence of a maturation culture, wherein the maturation culture comprises interleukin- 10 (IL-10), tumor necrosis factor-a (TNFa), IFNa, IFNy, and a TLR3 ligand.

56. The ex vzTO-generated antigen-presenting cell of claims 48-55, wherein the TLR3 ligand is polyinosinic:poly cytidylic acid (poly-I:C).

57. The ex vzvo-generated antigen-presenting cell of claims 48-55, wherein the IL-10 is at a concentration of 0.2-200 ng / ml.

58. The ex vzvc-generated antigen-presenting cell of claims 48-55, wherein the TNFa is at a concentration of 0.2-200 ng / ml.

59. The ex vzvo-generated antigen-presenting cell of claims 48-55, wherein the IFNa is at a concentration of 1-10,000 U / ml.

60. The ex vzvc-generated antigen-presenting cell of claims 48-55, wherein the IFNy is at a concentration of 1-10,000 U / ml.

61. The ex vzvo-generated antigen-presenting cell of claims 48-56, wherein the poly-I:C is at a concentration of 0.2 ng / ml-500 pg / ml.

62. The ex vi vo-generated antigen-presenting cell of claims 48-61, wherein the said ex vivo- generated antigen-presenting cell produces high levels of interleukin- 12 (IL-12p70), interleukin- 18 (IL- 18), C-X-C motif chemokine ligand 9 (CXCL9, )C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 11 (CXCL11), chemokine ligand 5 (CCL5), or chemokine ligand 19 (CCL19) in response to CD40 ligand (CD40L) stimulation as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-0 signaling.

63. The ex vzvo-generated antigen-presenting cell of claims 48-62, wherein the said ex vivo- generated antigen-presenting cell produces at least 50% or more of IL-12p70, IL-18, CXCL9, CXCL10, CXCL11, CCL5, or CCL19 in response after 24 hours of stimulation to CD40L as compared to standard antigen-presenting cell not exposed to the inhibitors of TGF-0 signaling.

64. A cancer vaccine comprising an ex vzvo-generated antigen-presenting cell of claims 48-63, loaded ex vivo with at least one or more tumor antigen peptides.

65. An activated antigen specific T-cell, comprising activating the antigen specific T-cell with an ex vzvc-generated antigen-presenting cell of claims 48-63.

66. The activated antigen specific T-cell of claims 48-65, wherein the activated antigen specific T-cell is administered intratumorally.

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