Compositions and methods for producing dendritic cell-based vaccines with enhanced efficacy

Abstract

Compositions and methods for improving the efficiency and efficacy of dendritic cell (DC)-based vaccines, such as vaccines for the treatment of cancer, are described. The DC compositions invoke robust T cell responses even in immune-suppressive environments in tumor-bearing animals. Methods of making improved DC-based vaccines and methods of treating cancer using the DC-based compositions are also described.

Description

[0001] COMPOSITIONS AND METHODS FOR PRODUCING DENDRITIC CELL-BASED VACCINES WITH ENHANCED EFFICACY

[0002] CROSS REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 586,629, filed September 29, 2023, which is herein incorporated by reference in its entirety.

[0004] FIELD

[0005] This disclosure concerns methods of producing improved dendritic cell (DC)-based tumor vaccines by including an omega-3 lipid in the culture media for generating bone marrow-derived and monocyte- derived DCs. Also disclosed are DC-based vaccines produced by these methods and use of the DC-based vaccines in methods of treating cancer.

[0006] BACKGROUND

[0007] The unique ability of dendritic cells (DCs) to capture, process, and present antigens to T cells makes them attractive candidates for developing vaccines. Multiple strategies have been used to design an effective therapeutic DC-hased vaccine such as ex-vivo loading of DCs with immunogenic tumor antigens; genetic modification of DCs for enhanced antigen presentation; and delivery in combination with immune checkpoint inhibitors. However, none of these strategies have shown substantial benefits. One major obstacle to overcome is immune suppression by tumor-derived factors. Cellular lipid pools get altered during cancer progression and cancer cells tend to make saturated and monounsaturated lipids. Intracellular accumulation of these lipids can interfere with DC function. Therefore, cancer-specific changes in the cellular lipid pool can affect the outcome of DC cancer vaccine therapy.

[0008] SUMMARY

[0009] Disclosed herein are compositions and methods for improving the efficiency of dendritic cell (DC)- based vaccines, such as vaccines for the treatment of cancer. The DC vaccine compositions described herein invoke robust T cell responses even in immune- suppressive environments in tumor-bearing animals. The DC vaccines also show increased T cell dwell time, as well as enhanced T-cell activation and IFN-y production, thereby enhancing the potency of the DC-based vaccines. Methods of using the DC -based vaccines to treat cancer are also disclosed.

[0010] Provided herein are methods of producing DC -based tumor vaccines, wherein the DCs are produced from either bone marrow cells or monocytes (such as monocytes in a peripheral blood mononuclear cell (PBMC) sample). In some aspects, the method includes culturing bone marrow cells in culture media that includes granulocyte macrophage colony stimulating factor (GM-CSF) and an omega-3 lipid to produce immature DCs; and either (a) culturing the immature DCs with lipopolysaccharide (LPS) to produce mature DCs, and pulsing the mature DCs with a tumor antigen to produce mature DCs expressing the tumor antigen; or (b) transducing the immature DCs with a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen, and culturing the transduced immature DCs with LPS to produce mature DCs expressing the tumor antigen. In other aspects, the method includes culturing monocytes (such as monocytes in a PBMC sample) in culture media that includes GM-CSF, interleukin-4 (IL-4) and an omega-3 lipid to produce immature DCs; and either (a) culturing the immature DCs with LPS and interferon (IFN)-y lb to produce mature DCs, and pulsing the mature DCs with a tumor antigen to produce mature DCs expressing the tumor antigen; or (b) transducing the immature DCs with a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen, and culturing the transduced immature DCs with LPS and IFN-y lb to produce mature DCs expressing the tumor antigen. These methods lead to increased levels of omega-3 lipid in the plasma membrane of the DCs (“high omega-3 DCs”), which without being bound by theory, is believed to be responsible for the enhanced anti-tumor response exhibited by the DC-based vaccines produced according to these methods. In some aspects of these methods, the omega-3 lipid is docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a pro-resolvin lipid (e.g., RvDl, RvD2, RvEl, RvE4, PD-1, MR-1, MR-2 or 17-HDHA).

[0011] Also provided herein are DC -based vaccines produced by a method disclosed herein. In some aspects, the DC-based vaccines have an increased level of omega-3 lipid in their plasma membrane compared to DCs generated in the absence of omega-3 lipid. In some aspects, the DC -based vaccines exhibit increased T-cell dwell time, and enhanced T-cell activation and / or IFNy production compared to DCs generated in the absence of omega-3 lipid.

[0012] Further provided are methods of treating cancer in a subject. In some aspects, the method includes administering to the subject a DC-based vaccine disclosed herein, or a DC-based vaccine produced according to a method disclosed herein. In some examples, the DCs are autologous DCs (the bone marrow cells or monocytes used to generate the DCs were obtained from the subject to be treated). In other examples, the DCs are allogeneic. In some examples, the method further includes administering or providing one or more additional cancer therapies, such as administration of a checkpoint inhibitor. In some examples of the disclosed methods, the size or number of tumors in the subject is reduced by at least 25%, cachexia is reduced by at least 25%, and / or survival of the subject is increased by at least 25% as compared to a subject not receiving the DC -based vaccine or as compared to a subject receiving a DC-based vaccine, wherein the monocytes or bone marrow cells used to generate the DCs were not cultured with an omega-3 lipid.

[0013] Also herein are compositions that include isolated DCs, isolated monocytes, or isolated bond marrow cells and an omega-3 lipid. In some examples, the omega-3 lipid includes DHA, EPA or a pro- resolvin lipid. The pro-resolvin lipid can be, for example, resolvin DI (RvDl), resolvin D2 (RvD2), resolvin El (RvEl), resolvin E4 (RvE4), protectin DI (PD-1), maresin 1 (MR-1), maresin 2 (MR-2), or 17- hydroxy docosahexaenoic acid (17-HDHA). In some aspects, the isolated DCs, monocytes or bone marrow cells include a recombinant nucleic acid molecule, vector, or virus expressing a tumor antigen. In other aspects, the isolated DCs, monocytes or bone marrow cells have been pulsed with a tumor antigen (and thereby present the tumor antigen on the surface in the context of MHC). In some aspects, the isolated DCs, monocytes or bone marrow cells have an increased level of omega-3 lipid in their plasma membrane compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid.

[0014] The foregoing and other features of this disclosure will become more apparent from the following detailed description of several aspects which proceeds with reference to the accompanying figures.

[0015] BRIEF DESCRIPTION OF THE DRAWINGS

[0016] FIGS. 1A-1L: FAT-1 transgenic mice show improved overall survival, lower body mass, and protection against tumor growth compared to WT mice. (FIG. 1A). Schematic of FAT-1 transgenic mouse model. (FIG. IB) Kaplan Meier curve showing the survival probability of WT and FAT-1 naive animals (*p<0.02). (FIG. 1C) Comparison of body weight of WT and FAT-1 animals as they age (****p<0.0005). (FIG. ID) Tumor growth kinetics of TC-1 cell lines in FAT-1 and WT mice (*p<0.02). (FIG. IE) Kaplan- Meier survival curve for TC-1 tumor-bearing WT and FAT-1 mice (**p<0.009). (FIG. IF) Tumor growth kinetics of B16F10 melanoma cancer cell lines in WT and FAT-1 animals (***p<0.0001). (FIG. 1G) Kaplan-Meier survival curve for B16F10 tumor-bearing WT and FAT-1 mice (*p<0.002). (FIG. 1H) Percentage of tumor-free WT and FAT-1 mice post-inoculation (****p<0.005). Statistical analysis: Mann- Whitney test (unpaired / non-parametric comparing the difference in median). (FIG. II) Bar graph showing no difference in the frequency of major immune cell types in the spleen of WT and FAT-1 mice. (FIG. 1 J) Cartoon showing the experimental design of bone marrow dendritic cell (BMDC) culture and antigen presentation assay in vitro. (FIG. IK) Levels of interferon-gamma in supernatant from WT or FAT-1 BMDCs co-cultured with CD8+ T-cells from PMEL-1 mouse measured by ELISA (*p<0.008). (FIG. IL) Overlay of expression of major surface markers including activation and co-stimulatory markers (CD1 lb, CDllc, MHC Class II, CD40, CD80, CD86) between mature antigen-pulsed BMDCs derived from FAT-1 and WT mice. Statistical analysis: multiple comparisons were made using two-way ANOVA. The Sidek correction test was applied to correct for multiple comparisons.

[0017] FIGS. 2A-2I: FAT-1 derived BMDCs significantly reduce tumor burden in WT TC-1 tumorbearing mice. (FIG. 2A). Schematics for the genotypes of BM donor and antigen-pulsed DC recipient mice. (FIG. 2B) Tumor inoculation and vaccination schedule. (FIG. 2C) Tumor growth kinetic comparison of the vehicle (Veh) and WT DC-vaccinated tumor-bearing WT animals in the TC-1 tumor model. (FIG. 2D) Tumor growth kinetic comparison of the vehicle and FAT 1 DC-vaccinated tumor-bearing WT animals in the TC-1 tumor model. (FIGS. 2E-2G) Tumor growth rate between day 14 (before vaccine dose 2) and day 21 (before vaccine dose 3) for vehicle, WT DC-vaccinated or FAT-1 DC-vaccinated TC-1 tumor-bearing animals. (FIG. 2H) Kaplan-Meier survival curve for vehicle, WT DC-vaccinated, and FAT-1 DC-vaccinated TC-1 tumor-bearing animals. (FIG. 21) Comparison of body weight between vehicle, WT DC-vaccinated, and FAT- 1 DC-vaccinated TC-1 tumor-bearing animals. FIGS. 3A-3H: FAT-1 derived BMDCs significantly reduce tumor burden in WT B16F10 tumorbearing mice. (FIG. 3A) Tumor inoculation and vaccination schedule. (FIG. 3B) Kaplan-Meier survival curve for vehicle, WT DC-vaccinated, and FAT-1 DC-vaccinated B16F10 tumor-bearing animals. (FIG. 3C) Tumor growth kinetic comparison of the vehicle and WT DC-vaccinated tumor-bearing WT animals in the B16F10 tumor model. (FIG. 3D) Tumor growth kinetic comparison of the vehicle and FAT 1 DC-vaccinated tumor-bearing WT animals in the B16F10 tumor model. (FIGS. 3E-3G) Tumor growth rate between day 10 (before vaccine dose 2) and day 17 (before vaccine dose 3) for vehicle (FIG. 3E), WT DC-vaccinated (FIG. 3F), or FAT-1 DC-vaccinated (FIG. 3G) B16F10 tumor-bearing animals. (FIG. 3H) Proportion of mice with smaller tumors (<600 mm3in TC-1 and <500mm3in B16F10) in different DC vaccine groups in TC-1 and B16F10 tumor-bearing mice.

[0018] FIGS. 4A-4H: FAT-1 derived BMDCs induce a higher vaccine-specific response ex vivo. (FIG. 4A) Schematic showing the layout of peptide restimulation of whole-blood ex vivo. (FIGS. 4B, 4C) Bar graphs comparing the frequency of IFNy+ CD8 T-cells (FIG. 4B), and E7-tetramer+ INFy+ CD8 T cells (FIG. 4C) in overnight re-stimulation of whole-blood with E7 peptide vs vehicle control across different vaccination groups. (FIG. 4D) Bar graph comparing the frequency of PD1+ CD8 T cells in 48 hours re-stimulation of whole-blood with E7 peptide vs vehicle control across different vaccination groups. (FIGS. 4E, 4F, 4G) Simple regression analysis showing the correlation of the frequencies of IFNy+ CD8 T cells (FIG. 4E), E7- tetramer+IFNy + CD8 T cells (FIG. 4F), and PD-1+ CD8+ T cells (FIG. 4G) in peptide restimulation with tumor volumes in different vaccination groups. (*p<0.05, **p<0.005, ***p <0.005).

[0019] FIGS. 5A-5D: Bulk RNA seq analysis. (FIG. 5A) Schematic of the experimental design. (FIG. 5B) Principal component analysis (PCA) of the top 1000 most variant transcripts illustrates that the majority of variance separates BM from immature / mature samples (PCI), immature vs mature are separated on PC2 and FAT-1 vs WT are separated on PC3 (-1.5% of the variance in the top 1000 variant transcripts). Since the transcriptional state for each stage is so different, differential expression analysis was performed between the FAT1 and WT samples at each stage individually. (FIG. 50) Euler diagram summarizing the overlap among differentially expressed genes (DEGs) changed between FAT-1 and WT for at least one experimental stage. Solid black outlines designate DEGs up in FAT-1 compared to WT, while dashed outlines are DEGs down in FAT-1 compared to WT at the stage indicated.

[0020] FIGS. 6A-6F: FAT-1 and WT mice have different cytokine profiles, which may influence T-cell dwell time with BMDCs. (FIG. 6A) Dot blot showing the differences between the intensity for individual cytokines / chemokines (duplicate dots for each) between immature BMDCs from FAT-1 and WT mice. (FIG. 6B) Cytokine / chemokine profile in supernatant taken from immature BMDCs. (FIG. 6C) Cytokine / chemokine profile of serum from WT and FAT-1 naive mice. (FIG. 6D) Image showing the FAT-1 and WT BMDC co-culture with PMEL-1 CD8 T cells stained with viability dye (T cells are stained with CMFDA and DCs are stained with CMPTX). (FIG. 6E) Time spent by an antigen specific T-cell with an antigen loaded dendritic cell in an overnight co-culture assay. (FIG. 6F) Track length generated by T-cells during co-culture assay. (****p<0.0005). FIGS. 7A-7E: Omega-3 fatty acids and SPMs derived therefrom mimic the antigen presentation ability of FAT- 1 DCs. (FIG. 7A) Schematic of bone marrow-derived DC culture in the presence of DHA and resolvins. (FIGS. 7B-7E) Interferon-gamma production by the BMDCs grown in the presence of DHA (FIG. 7B), RvDl (FIG. 7C), RvEl (FIG. 7D), and RvE4 (FIG. 7E) when co-cultured with antigen-specific T-cells, measured by ELISA (**p<0.005).

[0021] FIG. 8: Naive WT mice vaccinated with E7 -pulsed FAT-1 DCs show significantly greater INFy+ CD8 T cells compared to WT DC and vehicle control.

[0022] FIG. 9: Antigen presentation assay to test the ability of DCs generated from WT mice in varying doses of SPMs.

[0023] DETAILED DESCRIPTION

[0024] I. Introduction

[0025] In addition to genetic and epigenetic changes, metabolic reprogramming plays an important role in tumor initiation and progression. A switch to de novo fatty acid (FA) synthesis, upregulation of fatty acid synthase enzyme (FASN), and FA acid uptake mechanisms are metabolic changes reported in several cancers. Also, cancer cells upregulate mechanisms to exclude specific FAs, such as docosahexaenoic acid (DHA), an omega-3 fatty acid, from their microenvironment. In addition to playing roles in physiology and diseases, including cancer, recent studies have linked high omega-3 levels with a decreased mortality risk in humans. However, their roles in promoting anti-tumor immunity are not well understood.

[0026] Tumor-specific immune suppressive factors remain the main challenge for the optimal functioning of anti-tumor immunity. Immune cells can take up the FA / lipids produced by cancer cells, severely limiting their functions. One recent study showed that PGE2, a bioactive lipid derived from arachidonic acid (AA), an omega-6 fatty acid, prevents optimal functioning of cDCl (conventional DCs type 1), preventing the orchestration of CTLs (cytotoxic T cells) for tumor clearance. An optimal CTL response requires the dendritic cells (DCs) to present tumor antigens efficiently. The ability of DCs to capture, process, and present antigens to T-cells makes them attractive candidates for developing vaccines. However, most DC vaccine treatments have not been effective in the clinic. Therefore, there is a need to develop methods / techniques to enhance the effectiveness of DC cancer vaccines for optimum CTL anti-tumor response. Altering the cellular lipid repertoire of immune cells, including DCs, profoundly affects their function. Disclosed herein is a direct role of omega-3 fatty acids in enhancing the antigen-presenting ability of bone marrow-derived DCs, which finding enables the generation of more effective cancer vaccines.

[0027] The studies disclosed herein utilized a mouse model expressing the C. elegans FAT-1 transgene, which converts omega-6 (n-6) to omega-3 (n-3) fatty acids, hence having high omega-3 levels. The data showed that antigen-pulsed DCs derived from the bone marrow cells of FAT- 1 mice significantly reduced tumor growth compared to those derived from the bone marrow of WT mice. In addition, FAT-1 DCs prevented cachexia and prolonged survival of tumor-bearing mice in both TC-1 and B16F10 murine tumor models. The FAT-1 DC vaccine elicited vaccine-specific responses in both tumor-bearing and naive mice. In vitro experiments showed that the FAT-1 DCs had better antigen presentation ability than the WT DCs, leading to an increased production of IFNyby the antigen- specific CD8+ T cells when co-cultured with antigen-pulsed FAT-1 DCs. There were no significant differences in the expression pattern of activation and co-stimulatory molecules on their surfaces. However, RNA sequencing of FAT-1 and WT DCs revealed several differentially expressed genes (DEGs) with potential novel roles in antigen presentation and DC-T- cell interaction.

[0028] It is further disclosed herein that the T-cell dwell time was significantly higher with antigen-pulsed FAT-1 DCs than with WT DCs. WT DCs generated in the presence of omega-3 fatty acids and specialized pro-resolving lipid mediators (SPMs) mimicked the antigen presentation ability of FAT- 1 DCs in vitro. These data indicate that the increased omega-3 fatty acids in the DCs (“high omega-3 DCs”) enhance their cross-presentation ability and have translational potential in immune modulation and enhancing DC-based cancer vaccine therapy.

[0029] II. Abbreviations

[0030] 17-HDHA 17-hydroxy docosahexaenoic acid

[0031] APC antigen presenting cell

[0032] BM bone marrow

[0033] BMDC bone marrow dendritic cell

[0034] CR complete regression

[0035] CTL cytotoxic T lymphocyte

[0036] CXCL chemokine (C-X-C motif) ligand

[0037] DC dendritic cell

[0038] DEG differentially expressed gene

[0039] DHA docosahexaenoic acid

[0040] ELISA enzyme-linked immunosorbent assay

[0041] EPA eicosapentaenoic acid

[0042] FA fatty acid

[0043] FASN fatty acid synthase enzyme fdr false discovery rate

[0044] GM-CSF granulocyte macrophage-colony stimulatory factor

[0045] IFN interferon

[0046] IL-4 interleukin-4

[0047] LPS lipopolysaccharide

[0048] MHC major histocompatibility complex

[0049] MR maresin

[0050] PCA principal component analysis

[0051] PD-1 protectin DI PF4 platelet factor 4

[0052] Rv resolvin

[0053] SPM pro-resolving lipid mediator

[0054] TCR T cell receptor

[0055] WT wild-type

[0056] III. Summary of Terms

[0057] Unless otherwise noted, technical terms are used according to conventional usage. Definitions of many common terms in molecular biology may be found in Krebs et al. (eds.), Lewin 's genes XII, published by Jones & Bartlett Learning, 2017. As used herein, the singular forms “a,” “an,” and “the,” refer to both the singular as well as plural, unless the context clearly indicates otherwise. For example, the term “a dendritic cell” includes singular or plural dendritic cells and can be considered equivalent to the phrase “at least one dendritic cell.” As used herein, the term “comprises” means “includes.” It is further to be understood that any and all base sizes or amino acid sizes, and all molecular weight or molecular mass values, given for nucleic acids or polypeptides are approximate, and are provided for descriptive purposes, unless otherwise indicated. Although many methods and materials similar or equivalent to those described herein can be used, particular suitable methods and materials are described herein. In case of conflict, the present specification, including explanations of terms, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. To facilitate review of the various aspects, the following explanations of terms are provided:

[0058] Adjuvant: A substance or vehicle that non-specifically enhances the immune response to an antigen. In some aspects, an adjuvant can include a suspension of minerals (alum, aluminum hydroxide, or phosphate) on which antigen is adsorbed; or water-in-oil emulsion, for example, in which antigen solution is emulsified in mineral oil (Freund incomplete adjuvant), sometimes with the inclusion of killed mycobacteria (Freund's complete adjuvant) to further enhance antigenicity (inhibits degradation of antigen and / or causes influx of macrophages). Adjuvants for use in the disclosed compositions can include, for example, the QS21 purified plant extract, Matrix M, AS01, MF59, and ALFQ adjuvants. Immunostimulatory oligonucleotides (such as those including a CpG motif) can also be used as adjuvants. Adjuvants include biological molecules (a “biological adjuvant”), such as costimulatory molecules. Exemplary adjuvants include IL-2, RANTES, GM-CSF, TNF-a, IFN-y, G-CSF, LFA-3, CD72, B7-1, B7-2, OX-40L, 4-1BBL and toll-like receptor (TLR) agonists, such as TLR-9 agonists. Additional adjuvants are known (see, e.g., Singh (ed.) Vaccine Adjuvants and Delivery Systems. Wiley-Interscience, 2007).

[0059] Administration: The introduction of a composition into a subject by a chosen route. For example, if the chosen route is intravenous, the composition is administered by introducing the composition into a vein of the subject. Exemplary routes of administration include, but are not limited to, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, and intravenous), oral, intraductal, sublingual, rectal, transdermal, intranasal, vaginal and inhalation routes. Allogeneic: Refers to individuals, tissues or cells that are of the same species, but are antigenically distinct. AB allogeneic plasma is plasma obtained from healthy normal (allogeneic) donors with blood type AB. AB plasma lacks antibodies to both blood group A and blood group B, making it universally compatible in blood type at the plasma level. AB allogeneic plasma can be from a single AB donor or a pool of AB donors. The plasma is tested and confirmed to be negative for infectious pathogens, e.g., hepatitis A, B, C, and HIV. AB allogeneic plasma used in the compositions and method disclosed herein can be heat inactivated (HI AB) and used to culture monocytes into mature dendritic cells as an alternative to the use of autologous plasma (plasma from the patient) in the process of generating therapeutic dendritic cell (DC) vaccines.

[0060] Antigen presenting cells (APCs): A type of cell that displays antigens complexed with major histocompatibility complex (MHC) proteins on their surface. Professional APCs are very efficient at internalizing antigen, processing it and then displaying small pieces of the antigen (peptides) bound to a MHC molecule on the cell membrane surface. The three main types of professional APCs include DCs, macrophages and certain B cells. DCs have the broadest range of antigen presentation and are the most important APC in processing antigens for presentation to T cells, which recognize antigen-MHC complexes using their T cell receptors.

[0061] Autologous: Derived from the same individual. In the context of the present disclosure, “autologous” DCs used for treatment of a subject are DCs (or bone marrow cells or monocytes) originally obtained from the subject.

[0062] Cachexia: A metabolic syndrome that causes muscle loss and weight loss, often due to an underlying chronic illness, such as cancer, heart failure and chronic obstructive pulmonary disease. In cancer patients, the prevalence of cachexia ranges from 40% at diagnosis to 70% in advanced disease. Other symptoms of cachexia include loss of appetite, fatigue and lack of strength. Cachexia is also referred to as wasting syndrome.

[0063] Cancer: A malignant neoplasm that has undergone characteristic anaplasia with loss of differentiation, increased rate of growth, invasion of surrounding tissue, and is capable of metastasis. For example, prostate cancer is a malignant neoplasm that arises in or from prostate tissue, and breast cancer is a malignant neoplasm that arises in or from breast tissue (such as a ductal carcinoma). Metastatic cancer is a cancer at one or more sites in the body other than the site of origin of the original (primary) cancer from which the metastatic cancer is derived.

[0064] Checkpoint inhibitor: A class of drugs (typically antibodies) that target proteins that play a role in suppressing the immune response. Immune checkpoint inhibitors include, for example, antibodies or other inhibitors that target CTLA-4, PD-1, PD-L1, TIM-3 or LAG-3.

[0065] Chemotherapeutic agent: Any chemical agent with therapeutic usefulness in the treatment of diseases characterized by abnormal cell growth. Such diseases include tumors, neoplasms, and cancer. In one aspect, a chemotherapeutic agent is a radioactive compound. Exemplary chemotherapeutic agents that can be used with the methods provided herein are disclosed in Slapak and Kufe, Principles of Cancer Therapy, Chapter 86 in Harrison's Principles of Internal Medicine, 14th edition; Perry el al., Chemotherapy, Ch. 17 in Abeloff, Clinical Oncology 2nded., © 2000 Churchill Livingstone, Inc; Baltzer, L., Berkery, R. (eds.) Oncology Pocket Guide to Chemotherapy, 2nd ed. St. Louis, Mosby-Year Book, 1995; Fischer, D.S., Knobf, M.F., Durivage, H.J. (eds): The Cancer Chemotherapy Handbook, 4th ed. St. Louis, Mosby-Year Book, 1993). In one example, a chemotherapeutic agent is a biologic, such as a therapeutic antibody (e.g., therapeutic monoclonal antibody), such as an antibody targeting a tumor antigen, as well as other anti-cancer antibodies, such as anti-PDl or anti-PDLl (e.g., pembrolizumab and nivolumab), anti-CTLA4 (e.g., ipilimumab), anti-EGFR (e.g., cetuximab), anti-VEGF (e.g., bevacizumab), or combinations thereof (e.g., anti-PD-1 and anti-CTLA-4). Combination chemotherapy is the administration of more than one agent to treat cancer. One example is the administration of a DC-based vaccine disclosed herein used in combination with a radioactive, biological, or chemical compound, or combinations thereof.

[0066] Contacting: Placement in direct physical association; includes both in solid and liquid form. Dendritic cells (DCs): The principle professional antigen presenting cells (APCs) involved in primary immune responses. They are potent activators of T helper cell responses because as part of their composition, they express co-stimulatory molecules on their cell surface. Their major function is to obtain antigen in tissues, migrate to lymphoid organs and present the antigen to activate T cells. Immature dendritic cells originate in the bone marrow and reside in the periphery as immature cells. Dendritic cell sub-types include plasmacytoid dendritic cells and myeloid dendritic cells.

[0067] Docosahexaenoic acid (DHA): An omega-3 fatty acid with the chemical formula C22H32O2. DHA is the primary structural component of the human brain, retina, and skin.

[0068] Eicosapentaenoic acid (EPA): An omega-3 fatty acid with the chemical formula C20H30O2. EPA is a polyunsaturated fatty acid that acts as a precursor for prostaglandin-3, thromboxane-3, and leukotriene-5 eicosanoids.

[0069] Granulocyte-macrophage colony-stimulating factor (GM-CSF): An immunomodulatory cytokine that promotes production of hematopoietic cells such as granulocytes and macrophages. GM-CSF also has antiviral effects.

[0070] Immune response: A response of a cell of the immune system, such as a B cell, T cell, or monocyte, to a stimulus. In one aspect, the response is specific for a particular antigen (an “antigen-specific response”). In one aspect, an immune response is a T cell response, such as a CD4+response or a CD8+response. In another aspect, the response is a B cell response, and results in the production of specific antibodies.

[0071] Inhibiting, ameliorating, or treating a disease: “Inhibiting” a condition refers to inhibiting the full development of a condition or disease, for example, cancer or a tumor. Inhibition of a condition occurs within the spectrum from partial inhibition to substantially complete inhibition of the disease. In some examples, the term “inhibiting” refers to reducing or delaying the onset or progression of a condition. “Ameliorating” refers to the reduction in the number or severity of signs or symptoms of a disease, such as cancer. “Treating” refers to a therapeutic intervention that decreases or inhibits a sign or symptom of a disease or pathological condition after it has begun to develop, such as a reduction in tumor size or tumor burden. A subject to be administered an effective amount of the disclosed modified immune cells can be identified by standard diagnosing techniques for such a disorder, for example, presence of the disease or disorder or risk factors to develop the disease or disorder.

[0072] Interferon-g (IFN-y) lb: A recombinant form of the endogenous cytokine IFN-y that can be used to treat certain infections and osteopetrosis. IFNy-lb is 140 amino acids in length and is modified with an N- terminal methionyl group. Upon administration, IFNy-lb binds to IFNycell surface receptors (IFNGRs), activates IFNy-stimulated signaling pathways, including the JAK-STAT pathway, and leads to the transcription of multiple IFNy-controlled genes, the expression of which may activate certain components of the immune system. IFNy activates natural killer (NK) cells, macrophages, and cytotoxic T lymphocytes (CTLs), stimulates antibody-dependent cellular cytotoxicity (ADCC) and induces the expression of major histocompatibility complex (MHC) molecules, which leads to an increase in antigen presentation, including tumor-associated antigens (TAAs), to the immune system, and modulates the expression of certain pro- inflammatory cytokines by antigen-presenting cells (APCs). Altogether, this increases tumor cell killing.

[0073] Interleukin-4 (IL-4): A cytokine that plays a role in several biological processes, such as inducing differentiation of naive helper T cells, stimulation of activated B cell and T cell proliferation, differentiation of B cells into plasma cells, activation of macrophages, modulating DC response to interferons, and triggering differentiation of monocytes into DCs.

[0074] Isolated: An “isolated” biological component (such as a polynucleotide, protein, or cell) has been substantially separated, produced apart from, or purified away from other biological components (e.g., other cells, chromosomal and extra-chromosomal DNA and RNA, and proteins). Cells, polynucleotides, and proteins that have been isolated specifically include cells, nucleic acid molecules, and proteins purified by standard purification methods. The term also embraces nucleic acid molecules and proteins prepared by recombinant expression or production in a host organism, as well as chemically-synthesized nucleic acid molecules.

[0075] Lipopolysaccharide (LPS): A large glycoconjugate that is a major component of the outer membrane of gram-negative bacteria. LPS is toll-like receptor 4 (TLR4) agonist with immunostimulatory activity.

[0076] Lung tumor: Any malignant tumor found in tissues of the lung. Lung tumors include, but are not limited to, small cell lung cancer, non-small cell lung cancer, large cell carcinoma, lung adenocarcinoma, squamous cell carcinoma, mesothelioma, and lung carcinoid tumors.

[0077] Major histocompatibility complex (MHC) Class I: MHC class I molecules are heterodimers formed from two non-covalently associated proteins, the HLA heavy chain and 02 -microglobulin. The HLA heavy chain includes three distinct domains, cxl, oc2 and cx3. The three-dimensional structure of the al and 2 domains forms the groove into which a peptide antigen fits for presentation to T cells. The a3 domain is an immunoglobulin-fold like domain that contains a transmembrane sequence, which anchors the a chain into the cell membrane of an antigen-presenting cell. MHC class I complexes, when associated with antigen (and in the presence of appropriate co-stimulatory signals), stimulate CD8+ cytotoxic T cells, which function to kill any cell which they specifically recognize.

[0078] Mammal: This term includes both human and non-human mammals. Similarly, the term “subject” includes both human and veterinary subjects (such as cats, dogs, cows, and pigs) and rodents (such as mice and rats).

[0079] Melanoma: A form of cancer that originates in melanocytes (cells that make the pigment melanin). Melanocytes are found primarily in the skin, but are also present in the bowel and eye. Melanoma in the skin includes superficial spreading melanoma, nodular melanoma, acral lentiginous melanoma, and lentigo maligna (melanoma). Any of the above types may produce melanin or can be amelanotic. Similarly, any subtype may show desmoplasia (dense fibrous reaction with neurotropism) which is a marker of aggressive behavior and a tendency to local recurrence. Other melanomas include clear cell sarcoma, mucosal melanoma and uveal melanoma.

[0080] MHC Class II molecules: Heterodimeric proteins comprised of an a chain, which includes an ccl region and an oc2 region, and a [3 chain, which includes a [31 region and a [32 region. The ccl and [31 regions form the peptide -binding region, and the oc2 and [32 regions form an immunoglobulin-like domain. MHC Class II molecules present extracellular antigens and stimulate helper T cells to produce cytokines to stimulate the immune response.

[0081] Monocyte: A mononuclear, phagocytic leukocyte that is normally found in lymph nodes, spleen, bone marrow and loose connective tissue. Monocytes constitute 3-7% of the leukocytes in the circulating blood. Under appropriate culture conditions, monocytes from the peripheral blood can differentiate into dendritic cells.

[0082] Omega-3 lipids: Polyunsaturated fatty acids (PUFAs) with a double bond three carbons from the end of their acyl tail. They are also known as n-3 fatty acids. Omega-3 lipids play an important role in cell structure and function, and they support the brain, heart, lungs, blood vessels, and immune system. Exemplary omega-3 lipids include, but are not limited to, DHA, EPA, alpha-linolenic acid (ALA) and pro- resol vin lipids.

[0083] Pharmaceutically acceptable carriers: The pharmaceutically acceptable carriers of use are conventional. Remington: The Science and Practice of Pharmacy. 22nded. , London, UK: Pharmaceutical Press, 2013, describes compositions and formulations suitable for pharmaceutical delivery of the disclosed compositions (such as DCs).

[0084] In general, the nature of the carrier will depend on the particular mode of administration being employed. For instance, parenteral formulations usually include injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle. For solid compositions (e.g.. powder, pill, tablet, or capsule forms), conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate. In addition to biologically neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, added preservatives (such as non-natural preservatives), and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate. In particular examples, the pharmaceutically acceptable carrier is sterile and suitable for parenteral administration to a subject for example, by injection. In some aspects, the active agent and pharmaceutically acceptable carrier are provided in a unit dosage form such as a pill or in a selected quantity in a vial. Unit dosage forms can include one dosage or multiple dosages (for example, in a vial from which metered dosages of the agents can selectively be dispensed).

[0085] Pro-resolvin lipids: Omega-3 fatty acid-derived bioactive lipids. Exemplary pro-resolvin lipids include, but are not limited to, resolvin DI (RvDl), resolvin D2 (RvD2), resolvin El (RvEl), resolvin E4 (RvE4), protectin DI (PD-1), maresin 1 (MR-1), maresin 2 (MR-2), and 17-hydroxy docosahexaenoic acid (17-HDHA). The omega-3 fatty acid DHA is the precursor of 17-HDHA, PD-1, MR-1, MR-2, and the D- series resolvin lipids. EPA is the precursor of the E-series resolvin lipids (see, e.g., Valdes et al., Scientific Reports 7: 10748, 2017; and Serhan et al., Annu Rev Pathol 3:279-312, 2008).

[0086] Pulsed (with antigen): Refers to incubating or culturing cells, such as dendritic cells (DCs), in the presence of an antigen, resulting in display of the antigen on DCs in the context of MHC molecules.

[0087] Recombinant: A recombinant nucleic acid, protein or virus is one that has a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two otherwise separated segments of sequence. This artificial combination can be accomplished, for example, by chemical synthesis or by the artificial manipulation of isolated segments of nucleic acids, for example, by genetic engineering techniques.

[0088] Subject: Living multicellular vertebrate organisms, a category that includes human and non-human mammals. In some aspects, the subject is a human.

[0089] T-cell dwell time: The time a T cell spends bound to a peptide-MHC complex, such as a peptide- MHC complex presented by dendritic cell or another antigen presenting cell.

[0090] Transduced: A transduced cell is a cell into which has been introduced a nucleic acid molecule by molecular biology techniques. As used herein, the term transduction encompasses all techniques by which a nucleic acid molecule might be introduced into such a cell, including transfection with viral vectors, transformation with plasmid vectors, and introduction of naked DNA by electroporation, lipofection, and particle gun acceleration.

[0091] Tumor, neoplasia, malignancy or cancer: A neoplasm is an abnormal growth of tissue or cells which results from excessive cell division. Neoplastic growth can produce a tumor. The amount of a tumor in an individual is the “tumor burden” which can be measured as the number, volume, or weight of the tumor. A tumor that invades the surrounding tissue and / or can metastasize to a location distant from the original / primary tumor is referred to as “malignant.” A “non-cancerous tissue” is a tissue from the same organ wherein the malignant neoplasm formed, but does not have the characteristic pathology of the neoplasm. Generally, noncancerous tissue appears histologically normal. A “normal tissue” is tissue from an organ, wherein the organ is not affected by cancer or another disease or disorder of that organ. A “cancer-free” subject has not been diagnosed with a cancer of that organ and does not have detectable cancer.

[0092] Exemplary solid tumors, such as cancers, that can be treated with the disclosed DC -based vaccines include metastatic solid tumors, such as breast carcinomas (e.g. lobular and duct carcinomas), sarcomas, carcinomas of the lung (e.g., non-small cell carcinoma, large cell carcinoma, squamous carcinoma, and adenocarcinoma), mesothelioma of the lung, colorectal adenocarcinoma, stomach carcinoma, prostatic adenocarcinoma, ovarian carcinoma (such as serous cystadenocarcinoma and mucinous cystadenocarcinoma), ovarian germ cell tumors, testicular carcinomas and germ cell tumors, pancreatic adenocarcinoma, biliary adenocarcinoma, hepatocellular carcinoma, bladder carcinoma (including, for instance, transitional cell carcinoma, adenocarcinoma, and squamous carcinoma), renal cell adenocarcinoma, endometrial carcinomas (including, e.g., adenocarcinomas and mixed Mullerian tumors (carcinosarcomas)), carcinomas of the endocervix, ectocervix, and vagina (such as adenocarcinoma and squamous carcinoma of each of same), tumors of the skin (e.g., squamous cell carcinoma, basal cell carcinoma, malignant melanoma, skin appendage tumors, Kaposi sarcoma, cutaneous lymphoma, skin adnexal tumors and various types of sarcomas and Merkel cell carcinoma), esophageal carcinoma, carcinomas of the nasopharynx and oropharynx (including squamous carcinoma and adenocarcinomas of same), salivary gland carcinomas, brain and central nervous system tumors (including, for example, tumors of glial, neuronal, and meningeal origin), tumors of peripheral nerve, soft tissue sarcomas and sarcomas of bone and cartilage, head and neck cancers, and lymphatic tumors (including B-cell and T- cell malignant lymphoma).

[0093] Exemplary hematologic cancers that can be treated with the disclosed DC-based vaccines include, but are not limited to, leukemias (e.g., acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, or adult T-cell leukemia), lymphomas (such as Hodgkin’s lymphoma and non-Hodgkin’s lymphoma), and myelomas.

[0094] Tumor antigen: A protein, peptide or other molecule that is expressed or overexpressed by a tumor cell. In some instances, tumor antigens are expressed only by tumor cells and not by non-tumor cells (generally referred to as “tumor-specific antigens” or “neoantigens”). In other instances, a tumor antigen is expressed to a higher level by a tumor cell compared to a non-tumor cell (generally referred to as a “tumor- associated antigen”). Exemplary tumor antigens include, but are not limited to, B-cell maturation antigen (BCMA), cancer antigen 125 (CA125), CD19, CD20, CD22, CD25, CD30, CD33, CD37, CD38, CD40, CD44, CD52, CD56, CD70, CD74, CD140, epithelial cell adhesion molecule (EpCAM), carcinoembryonic antigen (CEA), glycoprotein A33 (gpA33), Lewis Y, mesothelin, a-fetoprotein (AFP), IL-6, mucins (e.g., MUC1), platelet-derived growth factor receptor (PDGFR)-alpha, tumor-associated glycoprotein-72 (TAG- 72), carbonic anhydrase IX (CAIX), prostate-specific membrane antigen (PSMA), folate-binding protein, scatter factor receptor kinase, gangliosides (e.g., GD2, GD3 and GM2), glypican-1 (GPC1), GPC2, GPC3, cytokeratin, frizzled receptor, KRAS, prostate specific antigen (PSA), PSA-1, PSA-2, PSA-3, vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), VEGFR2, integrin aVp3, integrin a5 1, epidermal growth factor receptor (EGFR), epidermal growth factor-like protein-7 (EGFL7), fibronectin, hepatocyte growth factor (HGF), HER1, HER2, HER3, lysyl oxidase like-2 (LOXL2), MET, insulin growth factor-1 receptor (IGF1R), ephrin tyrosine kinase A3 (EPHA3), folate receptor (FR)- alpha, phosphatidyl-serine, syndecan-1, signaling lymphocytic activation molecule family 7 (SLAMF7; CD319), tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAILR1), TRAILR2, receptor activator of nuclear factor kappa-B ligand (RANKL), fibroblast activation protein (FAP), vimentin, tenascin, tyrosine-related protein-2 (TRP-2), MART-l / Melan-A, gplOO, adenosine deaminase-binding protein (ADAbp), cyclophilin b, colorectal associated antigen (CRC)-OO17-1A / GA733, carcinoembryonic antigen peptide-1 (CAP-1), CAP-2, etv6, AML1, melanoma antigen gene (MAGE)-Al, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-CI, MAGE-C2, MAGE-C3, MAGE-C4, MAGE-C5, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE- 6, GAGE-7, GAGE-8, GAGE-9, BAGE, RAGE, LAGE-1, non-steroidal anti-inflammatory drug-activated gene (NAG), N-acetylglucosaminyltransferase-V (GnT-V), multiple myeloma antigen 1 (MUM-1), cyclin dependent kinase 4 (CDK4), tyrosinase, p53, p21ras, receptor binding cancer antigen expressed on SiCo cells (RCAS1), E-cadherin, a-catenin, P-catenin, y-catenin, pl20-catenin (pl20ctn), preferentially expressed antigen in melanoma (PRAME), New York esophageal squamous cell carcinoma 1 (NY-ESO-1), cell division cycle 27 (CDC27), adenomatous polyposis coli protein (APC), fodrin, connexin 37, Ig-idiotype, pl5(PAF), gp75, human papilloma virus proteins (e.g., E6 and E7), EBV latent membrane protein 1 (LMP1), P1A, EBV-encoded nuclear antigen (EBNA)-l, brain glycogen phosphorylase, synovial sarcoma X chromosome breakpoint (SSX)-l, SSX-2 (HOM-MEL-40), SSX-4, SSX-5, SCP-1, Wilms tumor 1 (WTl), survivin, PR1, CA19-9, proteinase 3, elastase, cathepsin G, epithelial tumor antigen, fibroblast growth factor 5, and cancer / testis antigen 7 (CT-7).

[0095] Vaccine: A pharmaceutical composition that elicits a prophylactic or therapeutic immune response in a subject. In some cases, the immune response is a protective immune response. Typically, a vaccine elicits an antigen-specific immune response to an antigen of a pathogen, for example a viral pathogen, or to a cellular constituent correlated with a pathological condition (such as cancer). A vaccine may include a polynucleotide (such as a nucleic acid encoding an antigen), a peptide or polypeptide (such as an antigen), a virus, a cell (such as a dendritic cell) or one or more cellular constituents.

[0096] Vector: A nucleic acid molecule as introduced into a host cell, thereby producing a transformed host cell. A vector may include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector may also include one or more selectable marker genes and other genetic elements known in the art.

[0097] IV. Methods of Making DC-Based Vaccines

[0098] Provided herein are methods of producing DC-based vaccines, such as vaccines for the treatment of cancer. In the disclosed methods, the DC -based vaccines are generated from either bone marrow cells or monocytes (such as monocytes in a PBMC sample) that are cultured in the presence of an omega-3 lipid, which produces DCs with increased levels of omega-3 lipid in their plasma membranes (“high omega-3 DCs”). The DCs are either pulsed with a tumor antigen, or are transduced with a nucleic acid molecule, vector or virus that expresses a tumor antigen, both of which result in display of the tumor antigen on the DCs in the context of MHC molecules.

[0099] In some aspects, the method includes culturing bone marrow cells in culture media that includes granulocyte macrophage colony stimulating factor (GM-CSF) and an omega-3 lipid to produce immature DCs; and either (a) culturing the immature DCs with lipopolysaccharide (LPS) to produce mature DCs, and pulsing the mature DCs with a tumor antigen to produce mature DCs expressing the tumor antigen; or (b) transducing the immature DCs with a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen, and culturing the transduced immature DCs with LPS to produce mature DCs expressing the tumor antigen.

[0100] In some examples of this method using bone marrow cells, the cell culture media includes about 2 to about 40 ng / ml GM-CSF, such as about 10 to about 30 ng / ml GM-CSF, or about 15 to about 25 ng / ml GM- CSF, such as about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 or 40 ng / ml GM-CSF. In particular examples, the cell culture media includes about 20 ng / ml GM-CSF.

[0101] In some examples, the immature DCs are cultured with about I to about 20 ng / ml LPS or about 5 to about 15 ng / ml LPS, such as about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 ng / ml LPS. In particular examples, the immature DCs are culture with about 10 ng / ml LPS.

[0102] In some examples, the mature DCs are pulsed with about 5 |ig / ml to about 20 pg / ml of tumor antigen, or about 10 pg / ml to about 15 pg / ml of tumor antigen, such as about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 10 or 20 pg / rnl of the tumor antigen. The concentration of tumor antigen will vary depending the on antigen selected. An appropriate concentration of tumor antigen can be selected by a skilled person.

[0103] In some examples, the bone marrow cells are cultured with GM-CSF and the omega-3 lipid for about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days or about 10 days. In specific examples, the bone marrow cells are cultured with GM-CSF for 7 days.

[0104] In some examples, the culture media further includes pooled AB serum, autologous plasma, or allogeneic AB plasma.

[0105] In other aspects, the method includes culturing monocytes (such as monocytes in a PBMC sample) in culture media that includes GM-CSF, interleukin-4 (IL-4) and an omega-3 lipid to produce immature DCs; and either (a) culturing the immature DCs with LPS and interferon (IFN)-y lb to produce mature DCs, and pulsing the mature DCs with a tumor antigen to produce mature DCs expressing the tumor antigen; or (b) transducing the immature DCs with a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen, and culturing the transduced immature DCs with LPS and IFN-y lb to produce mature DCs expressing the tumor antigen.

[0106] In some examples of this method, the monocytes are derived from a peripheral blood mononuclear cell sample. In some examples, the culture media includes about 1000 lU / ml, about 1500 lU / ml, about 2000 lU / ml, about 2500 lU / ml, or about 3000 lU / ml of GM-CSF. In specific examples, the culture media includes 2000 lU / ml of GM-CSF.

[0107] In some examples, the culture media includes about 1000 lU / ml, about 1500 lU / ml, about 2000 lU / ml, about 2500 lU / ml, or about 3000 lU / ml IL-4. In specific examples, the culture media includes 2000 lU / ml IL-4.

[0108] In some examples, the culture media further includes keyhole limpet hemocyanin (KLH), or another immunogenic protein. In particular examples, the culture media includes about 5 mcg / ml, about 7.5 mcg / ml, about 10 mcg / ml, about 12.5 mcg / ml, or about 15 mcg / ml of KLH or another immunogenic protein.

[0109] In some examples, the culture media further includes pooled AB serum, autologous plasma, or allogeneic AB plasma.

[0110] In some examples, the immature DCs are cultured with about 5, about 10, about 15, about 20 or about 25 ng / ml of LPS. In specific examples, the immature DCs are cultured with about 15 ng / ml of LPS.

[0111] In some examples, the immature DCs are cultured with about 250 lU / ml, about 500 lU / ml, about 750 HJ / ml, about 1000 HJ / ml, about 1250 HJ / ml, or about 1500 lU / ml of IFN-y lb. In specific examples, the immature DCs are cultured with 1000 lU / ml of IFN-y lb.

[0112] In some aspects of these methods, the omega-3 lipid is docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a pro-resolvin lipid (e.g., RvDl, RvD2, RvEl, RvE4, PD-1, MR-1, MR-2 or 17-HDHA).

[0113] In some examples, the bone marrow cells or monocytes are cultured in the presence of DHA. In some examples, the bone marrow cells or monocytes cultured in the presence of about 0.1 pM to about 5 pM DHA, such as about 0.3 pM to about 4 pM DHA, or about 0.5 pM to about 3 pM DHA. In particular examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1 .5 pM, about 2 pM, about 2.5 pM or about 3 pM DHA. In specific nonlimiting examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 pM to about 5 pM DHA, such as about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM DHA.

[0114] In some aspects, the bone marrow cells or monocytes are cultured in the presence of EPA. In some examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 pM to about 5 pM EPA, such as about 0.3 pM to about 4 pM EPA, or about 0.5 pM to about 3 pM EPA. In particular examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM EPA. In specific nonlimiting examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 pM to about 5 pM EPA, such as about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM EPA. In some aspects, the bone marrow cells or monocytes are cultured in the presence of a pro-resolvin lipid (such as, but not limited to, RvDl, RvD2, RvEl, RvE4, PD-1, MR-1, MR-2 or 17-HDHA). In some examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 U.M to about 5 LlM of the pro-resolvin lipid, such as about 0.3 [J.M to about 4 LlM of the pro-resolvin lipid, or about 0.5 |XM to about 3 LlM of the pro-resolvin lipid. In particular examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 LlM, about 0.3 LlM, about 0.5 LlM, about 1 LlM. about 1.5 |1M, about 2 LlM, about 2.5 LlM or about 3 LlM of the pro-resolvin lipid. In specific non-limiting examples, the bone marrow cells or monocytes are cultured in the presence of about 0.1 LlM to about 5 Lt of the pro-resolvin lipid, such as about 0.1 JAM, about 0.3 p.M, about 0.5 ,nM, about 1 LlM, about 1.5 LlM. about 2 LlM, about 2.5 LtM or about 3 LlM of the pro-resolvin lipid.

[0115] In some aspects, the bone marrow cells or monocytes are autologous bone marrow cells or monocytes. In other aspects, the bone marrow cells or monocytes are allogenic bone marrow cells or monocytes (e.g., from a donor).

[0116] In some examples, the tumor antigen expressed by the DCs is B-cell maturation antigen (BCMA), cancer antigen 125 (CA125), CD19, CD20, CD22, CD25, CD30, CD33, CD37, CD38, CD40, CD44, CD52, CD56, CD70, CD74, CD140, epithelial cell adhesion molecule (EpCAM), carcinoembryonic antigen (CEA), glycoprotein A33 (gpA33), Lewis Y, mesothelin, a-fetoprotein (AFP), IL-6, mucins (e.g., MUC1), platelet- derived growth factor receptor (PDGFR)- alpha, tumor-associated glycoprotein-72 (TAG-72), carbonic anhydrase IX (CAIX), prostate-specific membrane antigen (PSMA), folate-binding protein, scatter factor receptor kinase, gangliosides (e.g., GD2, GD3 and GM2), glypican-1 (GPC1), GPC2, GPC3, cytokeratin, frizzled receptor, KRAS, prostate specific antigen (PSA), PSA-1, PSA-2, PSA-3, vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), VEGFR2, integrin aV 3, integrin a501, epidermal growth factor receptor (EGFR), epidermal growth factor-like protein-7 (EGFL7), fibronectin, hepatocyte growth factor (HGF), HER1, HER2, HER3, lysyl oxidase likc-2 (LOXL2), MET, insulin growth factor-1 receptor (IGF1R), ephrin tyrosine kinase A3 (EPHA3), folate receptor (FR)-alpha, phosphatidyl-serine, syndecan-1, signaling lymphocytic activation molecule family 7 (SLAMF7; CD319), tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAILR1), TRAILR2, receptor activator of nuclear factor kappa-B ligand (RANKL), fibroblast activation protein (FAP), vimentin, tenascin, tyrosine- related protein-2 (TRP-2), MART-l / Melan-A, gplOO, adenosine deaminase-binding protein (ADAbp), cyclophilin b, colorectal associated antigen (CRC)-OO17-1A / GA733, carcinoembryonic antigen peptide-1 (CAP-1), CAP-2, etv6, AML1, melanoma antigen gene (MAGE)-Al, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-CI, MAGE-C2, MAGE-C3, MAGE-C4, MAGE-C5, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, GAGE-9, BAGE, RAGE, LAGE-1, non-steroidal anti-inflammatory drug-activated gene (NAG), N-acetylglucosaminyltransferase-V (GnT-V), multiple myeloma antigen 1 (MUM-1), cyclin dependent kinase 4 (CDK4), tyrosinase, p53, p21ras, receptor binding cancer antigen expressed on SiCo cells (RCAS1), E-cadherin, a-catenin, 0-catenin, y-catenin, pl20-catenin (pl20ctn), preferentially expressed antigen in melanoma (PRAME), New York esophageal squamous cell carcinoma 1 (NY-ESO-1), cell division cycle 27 (CDC27), adenomatous polyposis coli protein (APC), fodrin, connexin 37, Ig-idiotype, p!5(PAF), gp75, human papilloma virus proteins (e.g., E6 and E7), EBV latent membrane protein 1 (LMP1), Pl A, EBV-encoded nuclear antigen (EBNA)-l, brain glycogen phosphorylase, synovial sarcoma X chromosome breakpoint (SSX)-l, SSX-2 (HOM-MEL-40), SSX-4, SSX-5, SCP-1, Wilms tumor 1 (WT1), survivin, PR1, CA19-9, proteinase 3, elastase, cathepsin G, epithelial tumor antigen, fibroblast growth factor 5, or cancer / testis antigen 7 ("CT-7).

[0117] V. Methods of Treating Cancer

[0118] Further provided are methods of treating cancer in a subject. In some aspects, the method includes administering to the subject a composition, a DC-based vaccine, or a DC-based vaccine produced according to a method disclosed herein.

[0119] In some aspects of the treatment methods, the DCs are generated from autologous bone marrow cells or monocytes (obtained from the subject to be treated). In other aspects, the DCs are generated from bone marrow cells or monocytes obtained from a donor.

[0120] In some aspects, the cancer is a solid tumor. Exemplary solid tumors / cancers include, but are not limited to, breast carcinomas (e.g. lobular and duct carcinomas), sarcomas, carcinomas of the lung (e.g., non-small cell carcinoma, large cell carcinoma, squamous carcinoma, and adenocarcinoma), mesothelioma of the lung, colorectal adenocarcinoma, stomach carcinoma, prostatic adenocarcinoma, ovarian carcinoma (such as serous cystadenocarcinoma and mucinous cystadenocarcinoma), ovarian germ cell tumors, testicular carcinomas and germ cell tumors, pancreatic adenocarcinoma, biliary adenocarcinoma, hepatocellular carcinoma, bladder carcinoma (including, for instance, transitional cell carcinoma, adenocarcinoma, and squamous carcinoma), renal cell adenocarcinoma, endometrial carcinomas (including, e.g., adenocarcinomas and mixed Mullerian tumors (carcinosarcomas)), carcinomas of the endocervix, ectocervix, and vagina (such as adenocarcinoma and squamous carcinoma of each of same), tumors of the skin (e.g., squamous cell carcinoma, basal cell carcinoma, malignant melanoma, skin appendage tumors, Kaposi sarcoma, cutaneous lymphoma, skin adnexal tumors and various types of sarcomas and Merkel cell carcinoma), esophageal carcinoma, carcinomas of the nasopharynx and oropharynx (including squamous carcinoma and adenocarcinomas of same), salivary gland carcinomas, brain and central nervous system tumors (including, for example, tumors of glial, neuronal, and meningeal origin), tumors of peripheral nerve, soft tissue sarcomas and sarcomas of bone and cartilage, head and neck cancers, and lymphatic tumors (including B-cell and T- cell malignant lymphoma).

[0121] In specific examples, the solid tumor is a lung tumor (for example, small cell lung cancer, non-small cell lung cancer, large cell carcinoma, lung adenocarcinoma, squamous cell carcinoma, mesothelioma, or lung carcinoid tumors). In other specific examples, the solid tumor is a melanoma (e.g., superficial spreading melanoma, nodular melanoma, acral lentiginous melanoma, lentigo maligna, clear cell sarcoma, mucosal melanoma or uveal melanoma).

[0122] In other aspects, the cancer is a hematologic cancer. Exemplary hematologic tumors include, but are not limited to, leukemia (for example, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, and adult T-cell leukemia), lymphomas (such as Hodgkin’s lymphoma and non-Hodgkin’s lymphoma), and myelomas).

[0123] In some examples, the cancer / tumor expresses one or more of the following antigens: BCMA, CA125, CD19, CD20, CD22, CD25, CD30, CD33, CD37, CD38, CD40, CD44, CD52, CD56, CD70, CD74, CD140, EpCAM, CEA, gpA33, Lewis Y, mesothelin, ALP, IL-6, a mucin (e.g., MUC1), PDGFR-a, TAG- 72, CAIX, PSMA, folate-binding protein, scatter factor receptor kinase, a ganglioside (e.g.. GD2, GD3 and GM2), GPC1, GPC2, GPC3, cytokeratin, frizzled receptor, KRAS, PSA, PSA-1, PSA-2, PSA-3, VEGF, VEGFR, VEGFR2, integrin aV 3, integrin a501, EGER, EGEL7, fibronectin, HGE, HER1, HER2, HER3, LOXL2, MET, IGF1R, EPHA3, FR-a, phosphatidyl-serine, syndecan-1, SLAMF7 / CD319, TRAILR1, TRAILR2, RANKL, FAP, vimentin, tenascin, TRP-2, MART-l / Melan-A, gplOO, ADAbp, cyclophilin b, CRC-0017-1A / GA733, CAP-1, CAP-2, etv6, AML1, MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-CI, MAGE-C2, MAGE-C3, MAGE-C4, MAGE-C5, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, GAGE-9, BAGE, RAGE, LAGE-1, NAG, GnT-V, MUM-1, CDK4, tyrosinase, p53, p21ras, RCAS1, E-cadherin, a-catenin, -catenin, y-catenin, p!20ctn, PRAME, NY-ESO-1, CDC27, APC, fodrin, connexin 37, Ig-idiotype, p!5(PAF), gp75, human papilloma virus proteins (e.g., E6 and E7), EBV LMP1, P1A, EBNA-1, brain glycogen phosphorylase, SSX-1, SSX-2 (HOM-MEL-40), SSX-4, SSX-5, SCP-1, WT1, survivin, PR1, CA19-9, proteinase 3, elastase, cathepsin G, epithelial tumor antigen, fibroblast growth factor 5, and CT-7.

[0124] In some examples of the disclosed methods, the size or number of tumors in the subject is reduced by at least 25%, at least 50%, at least 75%, at least 85%, at least 90%, at least 95% or at least 99%, cachexia is reduced by at least 25%, at least 50%, at least 75%, at least 85%, at least 90%, at least 95% or at least 99%, and / or survival of the subject is increased by at least 25%, at least 50%, at least 75%, at least 85%, at least 90%, at least 95% or at least 99%, as compared to a subject not receiving the DC -based vaccine or as compared to a subject receiving a DC-based vaccine, wherein the monocytes or bone marrow cells used to generate the DCs were not cultured with an omega-3 lipid.

[0125] In some aspects, the subject is a mammal. In some examples, the mammal is a human.

[0126] Administration of a DC-based vaccine composition disclosed herein can be accompanied by administration of other therapeutic treatments (such as surgical resection of a tumor, radiation therapy, chemotherapy, and administration of checkpoint inhibitors). In some aspects, a DC -based vaccine is used in combination with a checkpoint inhibitor, such as antibodies (or fragments thereof) that target CTLA-4, PD-1, PD-L1, LAG-3 or TIM-3. In some examples, the checkpoint inhibitor is an anti-CTLA-4 antibody. CTLA-4 is a protein receptor found on the surface of T cells. When activated, CTLA-4 is capable of suppressing the immune system response. Anti-CTLA-4 antibodies block the connection necessary to engage this protein, allowing the T cells to continue fighting cancer cells, rather than shutting down the immune response. In other examples, the checkpoint inhibitor is an anti-PD-1 antibody. The PD-1 checkpoint pathway is another pathway for a T cell immune response. When the PD-1 receptors on the surface of T cells connect with the PD-L1 molecules on the surface of cancer cells or other immune cells, signals are sent to the T cells to dampen the response. Anti-PD-1 drugs block the connection necessary to engage this protein, allowing the T cells to continue their response against the cancer cells. In yet other examples, the checkpoint inhibitor is an anti-PD-Ll antibody. Cancer cells have the ability to make certain molecules appear on the surface, including PDL-1 and PDL-2 of the PD-1 checkpoint pathway. Cancer cells may also cause immune cells near the cancer to express PD-L1. These molecules bind to the PD-1 on the T cells and turn them off. In other examples, the checkpoint inhibitor is an anti-LAG-3 antibody. LAG-3 is a negative regulator of T cell activity. In other examples, the checkpoint inhibitor is an anti-TIM-3 antibody. TIM-3 is an immune checkpoint receptor that inhibits the duration and magnitude of Thl and cytotoxic T cell responses, which dampens the anti-tumor immune response.

[0127] In some aspects, a checkpoint inhibitor is co-delivered locally with DC -based vaccine at the injection site. In other examples, the checkpoint inhibitor is co-formulated within the vaccine platform. Codelivery of a checkpoint inhibitor augments the vaccine response without the toxicity that is generally associated with the systemic delivery of these agents. Provided below is a list of exemplary checkpoint inhibitors that are either FDA approved or investigational agents:

[0128] In some aspects, the DC-based vaccine is used in combination with antibodies directed against immune-inhibitory cytokines, such as TGF-|3, IL- 13, IL- 10 or their receptors. Other combination treatments could include agents that inhibit or deplete regulatory T cells (for example, the IL-2 immunotoxin denileukin diftitox) and agents that inhibit or deplete myeloid suppressor cells (such as antibodies that target myeloid cell markers, for example anti-CDl lb, anti-Gr-1).

[0129] In some aspects, the DC-based vaccine is administered in combination with another therapeutic vaccine that expresses a tumor antigen also expressed by the patient’s tumor cells.

[0130] In some aspects, the DC-based vaccine is administered in combination with a natural killer T (NKT) cell agonist. In some examples, the NKT cell agonist is P-mannosylceramide (|3-ManCer; see U.S. Patent No. 8,835,613). In some cases, a subject is administered a composition that includes P-mannosylceramide or a salt or solvate thereof in a pharmaceutically acceptable carrier.

[0131] In some aspects, the DC-based vaccine is administered in combination with chemotherapy. In some examples, chemotherapy is administered first to reduce the metastatic tumor.

[0132] In some aspects, the DC-based vaccine is administered in combination with an oncolytic virus that specifically targets cancer cells, or another vector-based vaccine.

[0133] In some aspects, the DC-based vaccine is administered in combination with radiation therapy.

[0134] In other aspects, the DC -based vaccine is administered in combination with another anti-cancer agent. Any suitable anti-cancer agent can be administered in combination with the compositions disclosed herein. Exemplary anti-cancer agents include, but are not limited to, chemotherapeutic agents, such as, for example, mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, anti-survival agents, biological response modifiers, anti-hormones (e.g. anti-androgens) and anti-angiogenesis agents. Other anti-cancer treatments include radiation therapy and other antibodies that specifically target cancer cells.

[0135] Non-limiting examples of alkylating agents include nitrogen mustards (such as mechlorethamine, cyclophosphamide, melphalan, uracil mustard or chlorambucil), alkyl sulfonates (such as busulfan), nitrosoureas (such as carmustine, lomustine, semustine, streptozocin, or dacarbazine).

[0136] Non-limiting examples of antimetabolites include folic acid analogs (such as methotrexate), pyrimidine analogs (such as 5-FU or cytarabine), and purine analogs, such as mercaptopurine or thioguanine.

[0137] Non-limiting examples of natural products include vinca alkaloids (such as vinblastine, vincristine, or vindesine), epipodophyllotoxins (such as etoposide or teniposide), antibiotics (such as dactinomycin, daunorubicin, doxorubicin, bleomycin, plicamycin, or mitomycin C), and enzymes (such as L-asparaginase).

[0138] Non-limiting examples of miscellaneous agents include platinum coordination complexes (such as cis-diamine-dichloroplatinum II also known as cisplatin), substituted ureas (such as hydroxyurea), methyl hydrazine derivatives (such as procarbazine), and adrenocrotical suppressants (such as mitotane and aminoglutethimide) .

[0139] Non-limiting examples of hormones and antagonists include adrenocorticosteroids (such as prednisone), progestins (such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and magestrol acetate), estrogens (such as diethylstilbestrol and ethinyl estradiol), antiestrogens (such as tamoxifen), and androgens (such as testerone proprionate and fluoxymesterone). Examples of the most commonly used chemotherapy drugs include Adriamycin, Alkeran, Ara-C, BiCNU, Busulfan, CCNU, Carboplatinum, Cisplatinum, Cytoxan, Daunorubicin, DTIC, 5-FU, Fludarabine, Hydrea, Idarubicin, Ifosfamide, Methotrexate, Mithramycin, Mitomycin, Mitoxantrone, Nitrogen Mustard, Taxol (or other taxanes, such as docetaxel), Velban, Vincristine, VP-16, while some more newer drugs include Gemcitabine (Gemzar), Herceptin, Irinotecan (Camptosar, CPT-11), Eeustatin, Navelbine, Rituxan STI-571, Taxotere, Topotecan (Hycamtin), Xeloda (Capecitabine), Zevelin and calcitriol.

[0140] Non-limiting examples of immunomodulators that can be used include AS-101 (Wyeth-Ayerst Eabs.), bropirimine (Upjohn), gamma interferon (Genentech), GM-CSF (granulocyte macrophage colony stimulating factor; Genetics Institute), IU-2 (Cetus or Hoffman-LaRoche), human immune globulin (Cutter Biological), IMREG (from Imreg of New Orleans, La.), SK&F 106528, and TNF (tumor necrosis factor; Genentech).

[0141] Another common treatment for some types of cancer is surgical treatment, for example surgical resection of the cancer or a portion of it. Another example of a treatment is radiotherapy, for example administration of radioactive material or energy (such as external beam therapy) to the tumor site to help eradicate the tumor or shrink it prior to surgical resection. VI. Dendritic Cell (DC) Compositions

[0142] Provided herein are compositions that include isolated DCs, isolated monocytes, or isolated bone marrow cells and an omega-3 lipid. In some aspects, the omega-3 lipid includes docosahexaenoic acid (DHA), eicosapentaenoic acid (EP A), and / or a pro-resolvin lipid. The pro-resolvin lipid can be, for example, resolvin DI (RvDl), resolvin D2 (RvD2), resolvin El (RvEl), resolvin E4 (RvE4), protectin DI (PD-1), maresin 1 (MR-1), maresin 2 (MR- 2), or 17-hydroxy docosahexaenoic acid (17-HDHA).

[0143] In some aspects, the isolated DCs, monocytes or bone marrow cells include a recombinant nucleic acid molecule, vector, or virus expressing a tumor antigen (for example, the DCs, monocytes or bone marrow cells are transduced with a recombinant nucleic acid molecule, vector, or virus expressing a tumor antigen). The tumor antigen is expressed on the surface of the cell in the context of histocompatibility (MHC) molecules. In other aspects, the isolated DCs, monocytes or bone marrow cells have been pulsed with a tumor antigen, which allows for presentation of the tumor antigens on the surface of the cells via MHC molecules.

[0144] In some examples, the tumor antigen is B-cell maturation antigen (BCM A), cancer antigen 125 (CA125), CD19, CD20, CD22, CD25, CD30, CD33, CD37, CD38, CD40, CD44, CD52, CD56, CD70, CD74, CD140, epithelial cell adhesion molecule (EpCAM), carcinoembryonic antigen (CEA), glycoprotein A33 (gpA33), Lewis Y, mesothelin, a-fetoprotein (AFP), IL-6, mucins (e.g., MUC1), platelet-derived growth factor receptor (PDGFR)-alpha, tumor-associated glycoprotein-72 (TAG-72), carbonic anhydrase IX (CAIX), prostate-specific membrane antigen (PSMA), folate-binding protein, scatter factor receptor kinase, gangliosides (e.g., GD2, GD3 and GM2), glypican-1 (GPC1), GPC2, GPC3, cytokeratin, frizzled receptor, KRAS, prostate specific antigen (PSA), PSA-1, PSA-2, PSA-3, vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), VEGFR2, integrin aV 3, integrin a5 1, epidermal growth factor receptor (EGFR), epidermal growth factor-like protein-7 (EGFL7), fibronectin, hepatocyte growth factor (HGF), HER1, HER2, HER3, lysyl oxidase like-2 (LOXL2), MET, insulin growth factor- 1 receptor (IGF1R), ephrin tyrosine kinase A3 (EPHA3), folate receptor (FR)-alpha, phosphatidyl-serine, syndecan-1, signaling lymphocytic activation molecule family 7 (SLAMF7; CD319), tumor necrosis factor- related apoptosis-inducing ligand receptor 1 (TRAILR1), TRAILR2, receptor activator of nuclear factor kappa-B ligand (RANKL), fibroblast activation protein (FAP), vimentin, tenascin, tyrosine-related protein-2 (TRP-2), MART-l / Melan-A, gplOO, adenosine deaminase-binding protein (ADAbp), cyclophilin b, colorectal associated antigen (CRC)-OO17-1A / GA733, carcinoembryonic antigen peptide-1 (CAP-1), CAP- 2, etv6, AML1, melanoma antigen gene (MAGE)-Al, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-CI, MAGE-C2, MAGE-C3, MAGE-C4, MAGE-C5, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, GAGE-9, BAGE, RAGE, LAGE-1, non-steroidal anti-inflammatory drug-activated gene (NAG), N- acetylglucosaminyltransferase-V (GnT-V), multiple myeloma antigen 1 (MUM-1), cyclin dependent kinase 4 (CDK4), tyrosinase, p53, p21ras, receptor binding cancer antigen expressed on SiCo cells (RCAS1), E- cadherin, a-catenin, P-catenin, y-catenin, pl20-catenin (pl20ctn), preferentially expressed antigen in melanoma (PRAME), New York esophageal squamous cell carcinoma 1 (NY-ESO-1), cell division cycle 27 (CDC27), adenomatous polyposis coli protein (APC), fodrin, connexin 37, Ig-idiotype, pl5(PAF), gp75, human papilloma virus proteins (e.g., E6 and E7), EBV latent membrane protein 1 (LMP1), P1A, EBV- encoded nuclear antigen (EBNA)-l, brain glycogen phosphorylase, synovial sarcoma X chromosome breakpoint (SSX)-l, SSX-2 (HOM-MEL-40), SSX-4, SSX-5, SCP-1, Wilms tumor 1 (WT1), survivin, PR1, CA19-9, proteinase 3, elastase, cathepsin G, epithelial tumor antigen, fibroblast growth factor 5, or cancer / testis antigen 7 (CT-7).

[0145] In some aspects, the composition includes isolated DCs, monocytes or bone marrow cells and DHA. In some examples, the composition includes about 0.1 pM to about 5 pM DHA, such as about 0.3 pM to about 4 pM DHA, or about 0.5 pM to about 3 pM DHA. In particular examples, the composition includes about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM DHA. In specific non-limiting examples, the composition includes about 0.1 pM to about 5 pM DHA, such as about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM DHA.

[0146] In some aspects, the composition includes isolated DCs, monocytes or bone marrow cells and EPA. In some examples, the composition includes about 0.1 pM to about 5 pM EPA, such as about 0.3 pM to about 4 pM EPA, or about 0.5 pM to about 3 pM EPA. In particular examples, the composition includes about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM EPA. In specific non- limiting examples, the composition includes about 0.1 pM to about 5 pM EPA, such as about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM EPA.

[0147] In some aspects, the composition includes isolated DCs, monocytes or bone marrow cells and a pro- resolvin lipid (such as, but not limited to, RvDl, RvD2, RvEl, RvE4, PD-1, MR-1, MR-2 or 17-HDHA). In some examples, the composition includes about 0.1 pM to about 5 pM of the pro-resolvin lipid, such as about 0.3 pM to about 4 pM of the pro-resolvin lipid, or about 0.5 pM to about 3 pM of the pro-resolvin lipid. In particular examples, the composition includes about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM of the pro-resolvin lipid. In specific nonlimiting examples, the composition includes about 0.1 pM to about 5 pM of the pro-resolvin lipid, such as about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM or about 3 pM of the pro-resolvin lipid.

[0148] In some aspects, the isolated DCs, monocytes or bone marrow cells have an increased level of omega-3 lipid in their plasma membrane compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid.

[0149] In some aspects, the DCs, monocytes or bone marrow cells exhibit increased T-cell dwell time compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid. In some aspects, the DCs, monocytes or bone marrow cells exhibit enhanced T-cell activation and / or IFNy production compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid.

[0150] In some aspects, the composition further includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known and a skilled person can readily select a suitable pharmaceutically acceptable carrier for a DC-based composition.

[0151] In some aspects, the composition further includes an adjuvant. In some examples, the adjuvant is a suspension of minerals (alum, aluminum hydroxide, or phosphate) on which antigen is adsorbed; a water-in- oil emulsion, for example, in which antigen solution is emulsified in mineral oil (Freund incomplete adjuvant), sometimes with the inclusion of killed mycobacteria (Freund's complete adjuvant) to further enhance antigenicity (inhibits degradation of antigen and / or causes influx of macrophages). Adjuvants for use in the disclosed compositions can also include, for example, the QS21 purified plant extract, Matrix M, AS01 , MF59, and ALFQ adjuvants. Immunostimulatory oligonucleotides (such as those including a CpG motif) can also be used as adjuvants in the disclosed compositions. Adjuvants for use in the disclosed compositions can also include biological molecules, such as costimulatory molecules, for example IL-2, RANTES, GM-CSF, TNF-a, IFN-y, G-CSF, LFA-3, CD72, B7-1, B7-2, OX-40L, 4-1BBL, or toll-like receptor (TLR) agonists (e.g., TLR-9 agonists).

[0152] In some aspects, the composition further includes AB allogeneic plasma or pooled AB serum. In some examples, the AB allogeneic plasma or pooled AB serum used in the DC-based compositions is heat inactivated (HI).

[0153] In particular examples, the composition includes or further includes a pharmaceutically acceptable carrier, an adjuvant and AB allogenic plasma.

[0154] In some aspects, the composition further includes lipopolysaccharide (LPS), granulocytemacrophage colony-stimulating factor (GM-CSF), IL-4, or any combination thereof.

[0155] EXAMPLES

[0156] The following examples are provided to illustrate particular features of certain aspects of the disclosure, but the scope of the claims should not be limited to those features exemplified.

[0157] Example 1: Materials and Methods

[0158] This example describes the materials and experimental procedures for the studies described in Examples 2-8.

[0159] Reagents and antibodies

[0160] RPMI-1640 (Gipco-11875-093), FBS (Gibco-A52095-02), Glutamine, Penicillin / Streptomycin (Gibco- 15140-122), beta-mercaptoethanol (Gibco- 21985-023), D-PBS (Gibco (14190-144), R&D duo ELISA kit-IFN-y R&D (DY485), Milteyni mouse CD8a enrichment kit (Milteyni- 130-104-075), CMFDA cell tracker green dye (Invitrogen- C7025), CMPTX cell tracker red dye (C34552), DHA (Cayman Chemicals 90310), RvEl (Cayman Chemicals 10007279), RvE4 (Cayman Chemicals 29590), RvDl (Cayman Chemicals 872993-05-0), 17HDHA (Cayman Chemicals 10005099), PD1 (Cayman chemicals 10010390), MR-1 (Cayman chemicals 10878), LPS (Invitrogen- 50-112-2025), and GM-CSF (Preprotech 315-03).

[0161] Antibodies:

[0162] Mice, cell lines, and vaccination experimental design

[0163] C57BL / 6-Tg (CAG-fat-l)lJxk / J (Strain# 020097) (FAT-1) breeder pairs were purchased from Jackson Laboratory and bred in a pathogen-free facility for all experiments. PMEL-1 mice (Strain#005023) were purchased from Jackson Laboratory. C57BL / 6 WT mice were purchased from Charles River. Female mice, 10-12 weeks old were used for all tumor experiments. Animals were euthanized as the tumor approached 2 cm in length or at the first sign of distress, such as more than 15% body weight loss or morbid / hunched appearance.

[0164] B16F10 murine cancer cell lines were grown in complete RPML1640 (cRPMI) supplemented with 10% FBS, 100 lU / ml penicillin, 100 pg / nil streptomycin, and 2 mM L-glutamine at 37°C in humidified air with 5% CO2. TC-1 murine cancer cell lines were grown in RPMI 1640 supplemented with 10% FBS containing 100 lU / ml penicillin, 100 Ltg / ml streptomycin, 2 mM L-glutamine, 500 pl BME (betamercaptoethanol), and 100 pg / mL G418 (Gibco-10131035) at 37°C in humidified air with 5% CO2. For the B16F10 tumor model, 105cells / 100 pl were injected orthotopically into the skin. Five days post tumor inoculation, animals were injected with either hgplOO pulsed BMDC-FAT-1 or BMDC-WT DC vaccine intradermally away from the tumor injection site. For the TC-1 tumor model, 105cells / 100 pl were injected subcutaneously into the right flank. Seven days post TC-1 inoculation, animals were injected with either E7- peptide pulsed BMDC-FAT-1 or BMDC-WT DC vaccine intradermally away from the tumor injection site. PBS-injected tumor-bearing animals were used as a control group. For B16F10 and TC-1 groups, animals were injected weekly after the first vaccine dose, up to 4 doses.

[0165] BMDC generation and antigen presentation assay

[0166] Bone marrow cells were isolated from the femurs of 10- to 12-week old female FAT-1 or WT mice as described in Madaan et al. (J Biol Methods 1(1): 1, 2014). The cells were cultured in the cRPMI media in the presence of 20 ng / ml GM-CSF and one of the lipids (DHA, RvE4, or RvDl) for seven days. The immature BMDCs were frozen in cell freezing media and stored in liquid nitrogen. The cells were thawed the day before vaccination and plated in cRPMI media. Immature BMDCs were matured with 10 ng / ml LPS and pulsed with antigens (E7- 5 pg / ml, hgplOO 5 pg / ml) overnight. The next day, 2xl06cells / 100 pl were injected intradermally adjacent to the tumor.

[0167] Antigen presentation assay and ELISA

[0168] 20,000 immature BMDCs either from FAT-1 or WT mice were plated in 200 pl of cRPMI media in a flat bottom 96-well plate. The cells were matured and pulsed with 10 ng / ml LPS and 5 pg / ml hgplOO peptide overnight. The next day, the mature BMDCs were washed with PBS and co-cultured with 2x105CD8+ T-cells isolated from the female PMEL-1 mouse's spleen. The CD8+ T-cells were isolated by magnetic bead isolation using the Milteyni mouse CD8 T-cell enrichment kit. The manufacturer’s protocol was followed. The co-culture setup was incubated overnight at 37°C in humidified air with 5% CO2. The next day, supernatant from the co-culture was used to measure IFN-y levels using the R&D duo ELISA kit. The manufacturer’s protocol was followed.

[0169] Ex vivo peptide restimulation assay

[0170] Tumor-bearing WT animals vaccinated with either E7 -pulsed FAT-1 DCs or WT DCs were bled (mandibular bleed) 2-3 days after the second vaccine dose, and the blood was collected in EDTA tubes. The tubes were centrifuged at 1500 rpm for 10 minutes, and plasma was collected and frozen at -80°C until further use. The whole blood was plated in a 96-well U-bottom plate and restimulated with E7 -peptide ( 5 g / ml) overnight and for 48 hours. The cells were washed 3X with PBS, followed by surface staining with immune markers and FACS analysis to detect vaccine specificity and efficacy.

[0171] Cytokine array

[0172] Plasma was pooled from 6 independent biological replicates from either WT or FAT-1 animals and supernatants from BMDC culture were used to probe the array membrane from the R&D proteome profiler cytokine array kit (ARY006). The manufacturer’ s protocol was followed, and images were captured using the BioRad imager. The array image was analyzed and quantitated using Image J software. T-cell dwell time

[0173] The antigen presentation assay was set up as described above. The DC:T-cell ratio was kept at 1:2, and cells were plated in an eight-chamber glass slide from Ibidi (80841). Live imaging of the co-culture was carried out using a Nikon spinning disc confocal microscope overnight. The images were taken five minutes apart. Individual T-cells were tracked, and time spent in contact with one or more DCs was calculated per frame per T-cell in contact with DCs. Track length generated by individual T cells was calculated using the Imaris software.

[0174] Flow cytometry

[0175] Fluorescent protein-labeled antibodies against CD4, CD8, TCRfS, CD45, IFN-y, CDl lc, CDl lb, MHC class II, CD80, CD86, CD40 were used. HPV16 E749-57 peptide-loaded H2Dbtetramer was provided by the NIH tetramer core facility. For intracellular IFN-y staining, cells were fixed and permeabilized using Ebioscience FOXP3 Fixation / Permeabilization Kit (00-5523-00) following the manufacturer’s instructions. Cells were analyzed using a FACSymphony™ (BD Bioscience). FlowJo software was used for analyzing the data.

[0176] RNA sequencing

[0177] Total RNA from BM, immature, and mature BMDCs were harvested from three independent biological replicates from WT and FAT-1 mice and bulk RNA sequencing targeting 50M reads / samplc was performed on Novaseq SP. Raw reads were trimmed, aligned and normalized per CCBR’s RNA sequencing analysis pipeline (“RENEE”, github.com / CCBR / RENEE). Differentially expressed genes were determined using the DESeq2 R package (DESeq2_1.44.0) and considered significant with false discovery rate p < 0.05 and |FC| estimate > 2. Visualizations were generated in R using the following R package versions: euler diagram (eulerr_7.0.2), alluvial diagram (ggalluvial_0.12.5), heatmap (pheatmap_1.0.12), pathway enrichments (enrichR_3.2) and gene-concept networks (cnetplot function from enrichplot_1.24.0). Full code can be found at github.com / NCI-VB / berzofsky_tiwary_F ATl_DCbulk.

[0178] Example 2: High omega-3 fatty acids extend lifespan, prevent weight gain, and protect against tumor growth in mice

[0179] To demonstrate the direct impact of elevated omega-3 levels on lifespan extension and cancer protection, a genetically modified mouse model (FAT-1) was used. This model is a genetic knock-in of a FAT-1 desaturase gene from C. elegans. This gene can convert omega-6 fatty acid to omega-3 fatty acid by introducing double bonds to the carbon chain (FIG. 1 A). Hence, these animals have elevated levels of omega-3 fatty acids and a lower omega-6 to omega-3 ratio. The survival rates of naive FAT-1 and WT mice were compared over two years. The data show that the naive FAT-1 mice lived significantly longer than the WT mice (median overall survival 2.1 vs 1.4 years), indicating that high omega-3 levels significantly increase overall lifespan (FIG. IB). The body weight of FAT-1 and WT mice was monitored for up to 12 months and it was determined that post-weaning, FAT-1 mice weighed significantly less than WT mice (FIG. 1C, left panel), and their weights remained lower as they aged (FIG. 1C, middle and right panels). To investigate the effect of increased omega-3 levels on tumor growth, in vivo tumor growth assays were conducting using the TC-1 lung cancer model and the B16F10 orthotopic melanoma model. Significantly slower tumor growth was observed in FAT-1 mice compared to the WT mice across all tumor models (FIGS. ID and IF). Furthermore, the survival rate of mice with tumors was significantly longer in the FAT- 1 group compared to the WT group, as shown by the Kaplan-Meier survival curves for the TC-1 (FIG. IE) and Bl 6F 10 (FIG. 1G) tumor models.

[0180] In addition to the improved overall survival of tumor-bearing animals in the FAT-1 group compared to the WT group, a significant delay was observed in the onset of palpable tumors in the B16F10 tumor model (FIG. 1H). These data confirm that high omega-3 fatty acids can improve overall survival and suppress cancer progression in mice. Although there may be a direct effect of high omega-3 on tumor cells in FAT-1 mice, the high omega-3 fatty acids may also contribute to a robust and efficient anti-tumor immune response. Cross-presentation of tumor antigens by dendritic cells (DCs) is pivotal in generating an optimal anti-tumor CD8 T-cell response. Therefore, the antigen presentation ability of dendritic cells generated from bone marrow (BM) of FAT-1 mice was investigated and compared to DCs from the BM of WT mice.

[0181] Example 3: FAT-l-derived bone marrow dendritic cells (BMDCs) are efficient antigen presenters in vitro

[0182] It was first determined whether there were any differences in the frequencies of the immune cell sub-types between WT and FAT-1 mice at the basal level. To do so, the major immune markers on the splenocytes isolated from the spleen of naive FAT-1 and WT mice were evaluated. The data showed no difference in the frequencies of major immune cell subtypes in the spleen of naive FAT-1 and WT animals. (FIG. II).

[0183] Bone marrow (BM) cells were isolated from the femurs of FAT-1 and WT mice and BMDCs were generated from them to investigate the antigen presentation ability. BM cells were cultured in the presence of GM-CSF for seven days to generate immature BMDCs (FIG. 1J). After seven days, the immature BMDCs were matured with lipopolysaccharide (LPS) and pulsed with melanoma tumor antigen (hgplOO) overnight. The mature antigen-pulsed BMDCs were co-cultured with CD8 T-cells isolated from the spleen of PMEL-1 mice (which have a TCR specific for hgplOO) (FIG. 1 J). The level of interferon gamma (IFNy) was measured in the supernatant from the co-culture with peptide-pulsed FAT-1 BMDC and WT BMDCs by ELISA. The data indicate that FAT-1 BMDCs induced a higher IFNy response from the antigen-specific CD8 T-cells in the DC-CD8 T-cell co-culture assay than the WT BMDCs (FIG. IK). In addition, the expression level of the maturation and activation / costimulatory markers (CDl lb, CDllc, MHC Class II, CD40, CD80, CD86) on the surface of BMDCs from both FAT-1 and WT groups were compared. There were no differences in the expression levels of these activation / maturation markers (FIG. IL). These data indicate an improved ability of FAT-1 BMDCs, independent of the DC activation surface molecules, to induce an antigen-specific response by CD8 T-cells. Enhanced antigen presentation is an important attribute that defines the efficacy of DC -based therapeutic cancer vaccines. Therefore, it was further hypothesized that FAT-1 BMDCs may function as abetter DC vaccine and reduce tumor burden more efficiently than WT BMDCs in mice.

[0184] Example 4: FAT-l-derived DC vaccine significantly reduces tumor burden and cachexia and improves overall survival in murine tumor models

[0185] To evaluate the efficacy of the FAT-l-derived DC vaccine, the TC-1 tumor-bearing WT animals were vaccinated with either E7 antigen-pulsed FAT-1 DCs or WT DCs, as shown in FIG. 2A. The WT animals were vaccinated seven days after inoculating the tumor cells subcutaneously. FIG. 2B shows the timeline and frequency of vaccine doses. It was found that the FAT-1 DC vaccine significantly reduced the tumor burden in WT TC-1 tumor-bearing animals compared to the WT DC vaccine (FIGS. 2C and 2D). It was also observed that the maximum therapeutic efficacy was achieved between Day 14 and Day 21 (FIGS. 2E-2G), where almost 71% of the tumors were higher than the threshold of 600mm3in the WT DC vaccine group. Still, only 28% of the tumors in the FAT-1 DC vaccine group were in that range. Also, tumor regression was observed only in the FAT-1 DC group. In the control group, all the tumors crossed the threshold tumor volume. In addition to reducing the tumor burden, the FAT-1 DC vaccine also prolonged the survival of WT TC-1 tumor-bearing animals (FIG. 2H). In the FAT-1 group, three animals showed complete regression (CR) after the second dose of the vaccine. Out of those three animals, one remained tumor-free. However, the other two showed tumor relapse at a later timepoint (FIG. 2D). There were no CRs in the WT DC vaccine groups. It was observed that FAT-1 DC groups showed significantly less cachexia than WT DCs, a clinically favorable outcome (FIG. 21).

[0186] Similar results were also observed in the B16F10 melanoma model. The vaccination timeline was identical to the TC-1; however, in the B16F10 model, vaccination was started five days after the cells were inoculated due to the aggressive nature of this tumor model (FIG. 3 A). As in the TC-1 model, FAT-1 DCs significantly reduced the tumor burden in B16F10 tumor-bearing animals compared to the WT DCs (FIGS. 3C and 3D). In addition, the survival probability of the tumor-bearing mice in the FAT-1 DC vaccine group was significantly longer than that of the WT DC vaccine group (FIG. 3B). The maximum therapeutic benefit was observed between day 10 and day 17 (between 2ndand 3rddoses), where 100% of the tumors remained below the threshold of 500mm3in the FAT-1 DC group (FIG. 3G), compared with 60% in the WT DC group (FIG. 3F), and 46% in the vehicle group (FIG. 3E). Although no CRs existed in the B16F10 tumor model, partial tumor regression was observed only in the FAT-1 DC vaccine group. Together, these data suggest that DCs derived from the bone marrow of FAT-1 mice are superior to those derived from the WT mice at reducing the tumor burden in murine tumor models. Based on this observation, it was predicted that FAT-1 DCs induce a robust anti-tumor T-cell response in the tumor-bearing mice. The vaccine-specific T-cell response in the tumor-bearing and naive animals was further evaluated to test that hypothesis. Example 5: FAT-l-derived DC vaccines induce a better vaccine-specific response ex vivo

[0187] The data described herein suggest that the maximum therapeutic benefit of the vaccine is achieved between the second and third doses. Therefore, the vaccine-specific T-cell response from different groups was tested at the midpoint of the second and third vaccine doses (FIG. 4A). To do so, tumor-bearing vaccinated animals were bled and an ex vivo antigen re-stimulation assay was performed by pulsing the whole blood with the E7 peptide overnight and for 48 hours. The overnight ex vivo restimulation of the whole blood showed that FAT- 1 DC vaccinated animals had a significantly higher IFNy+ CD8 T cell frequency than WT DC vaccinated and vehicle control groups (FIG. 4B). Additionally, the frequency of E7- tetramer+ CD8 T cells that were also positive for IFNy was significantly higher in the FAT-1 DC group compared to the WT DC and the vehicle groups (FIG. 4C). Higher frequencies of IFNy+ CD8 T-cells significantly correlated with smaller tumor volumes in FAT-1 DCs compared to WT DCs (FIG. 4F). Also, there was a significant correlation between E7-tetramer+ IFNy + CD8 T-cells and smaller tumor size in the FAT-1 DC group compared to the WT DC group (FIG. 4G). In the 48-hour restimulation assay, the frequency of PD-1+ CD8 T-cells was much higher in the FAT-1 DC group than in the WT DC and the vehicle groups (FIG. 4D). Although not significant, there was a trend toward a high frequency of PD-1+ CD8 T-cells with smaller tumors in the FAT-1 DC group (FIG. 4H). In addition to tumor-bearing animals, the vaccine-specific CD8-T cell response was tested in the WT tumor naive animals. A significantly higher frequency of INFy+CD8 T-cells were found in the FAT-1 DC vaccinated naive group compared to the WT DC group (FIG. 8). This data suggests that FAT-1 DCs are significantly better at inducing a vaccine-specific CD8 T-cell response. Even though the co-stimulatory and activation molecules such as CD80, CD86, MHCII, and CD40 were expressed at similar levels between FAT-1 and WT BMDCs (FIG. 1H), the FAT-1 DCs show a remarkable effect on suppressing tumor growth in vivo and enhanced vaccine efficacy ex vivo. Therefore, bulk RNA sequencing was performed and the transcriptional changes between the FAT-1 and WT DCs was analyzed to investigate their differences further.

[0188] Example 6: Bulk RNA seq analysis reveals differentially expressed genes (DEGs) in FAT-1 and WT in BM, immature, and mature BMDCs

[0189] Since no differences were detected in the activation and maturation molecular markers between the FAT-1 and WT mature BMDCs, bulk RNA-sequencing (RNA-seq) was performed to evaluate their transcriptional profiles. Bone marrow and immature BMDCs were also included to assess the temporal changes as the cells differentiate from stem cells (bone marrow) to terminally differentiated dendritic cells state. FIG. 5A shows the study design for bulk RNA sequencing. Principal component analysis (PCA) validated robust differentiation from BM cells to BMDCs as most of the variance in the top 1000 most variant transcripts (>84%) separated immature / mature BMDCs from the BM precursors (FIG. 5B, PCI). A further 11% of the variation cleanly separated immature from mature BMDCs (FIG. 5B, PC2), indicating the transcriptional response to LPS treatment transcends the WT / FAT-1 genotypes. However, PC3 (1.5% of the variation in the top 1000 most variant transcripts) cleanly separated FAT-1 from the WT samples regardless of the differentiation stage. Thus, pair-wise differential expression was performed between FAT-1 and WT samples at each differentiation stage separately (FIG. 8A) and significant differentially expressed genes (DEGs) were overlaid to establish shared and stage-specific signatures (FIGS. 5C and 5D). While most DEGs were up or downregulated only at a specific differential stage (FIG. 5C), 16 upregulated DEGs and 5 downregulated DEGs were shared across all three stages and several more were shared between the intermediate stage immature BMDCs and either precursor BM (shared across “early” timepoints) or mature BMDCs (shared across “late” timepoints). These changes are likely caused by high omega-3 levels in the FAT-1 mice. Pathway enrichment for DEGs upregulated in FAT-1 compared with WT BM included TNF- alpha signaling, IL- 18 signaling, and hypoxia. Chemokine activity and IL-6 / JAK-STAT3 signaling were upregulated in FAT-1 immature BMDCs. At the same time, cell-substrate junction and endosomal membrane pathways were enriched in DEGs downregulated in FAT-1 immature BMDCs. DEGs upregulated in FAT-1 mature BMDCs were enriched for G alpha signaling events.

[0190] Example 7: RNA seq identifies candidate genes with the potential to influence immune cell recruitment and synapse formation between DCs and T-cells, affecting T-cell dwell time

[0191] CXLC2 and S 1 pr 1 were among the top upregulated DEGs in the mature FAT-1 DCs compared to the WT DCs. Several other chemokines, namely CCL2, CCL7, CXCL2, and PF4 / CXCL4, were upregulated in immature BMDCs, and Pf4 / CXCL4 was upregulated in both mature and immature BMDCs. The expression of these genes was further corroborated at the protein level by performing a cytokine array (FIG. 6A) from the supernatant taken from immature BMDCs derived from WT and FAT-1 and serum isolated from WT and FAT-1 mice. The data showed several of these cytokines present in FAT-1 BMDC culture supernatant and FAT-1 mouse serum (FIGS. 6B-6C). In addition, many genes potentially affecting synapses and dendrite formation in non-DC cell types, such as NBEA, DACH1, and WWC2, were among the top DEGs in FAT- 1 DCs.

[0192] It was hypothesized that with the expression of the genes with potential for regulating synapse and chemokines / cytokines that may serve as attractants for immune cells, it is likely that T-cells are spending more time in contact with FAT-1 DCs, and hence an increase in T-cell dwell time. To test that hypothesis, live imaging was performed of the co-culture of PMEL-1 CD8 T-cells with hgplOO peptide-pulsed mature BMDCs derived from WT or FAT-1 mice. The antigen-specific CD8-T cells spent more time with FAT-1 DCs than the WT DCs. At any given time, more CD8-T cells were in contact with DCs in the FAT-1 group than in the WT group (FIG. 6D). The total time spent by individual T-cells in contact with a DC was calculated for both groups and it was found that T-cells spent significantly and substantially (> 10-fold) longer time with FAT-1 DCs than WT-DCs. On the other hand, most of the DC-T cell interactions were short-lived in the WT DC group (FIG. 6E). Consistent with that, the track length generated by each T-cell per image frame was much longer in the WT DC groups compared to the FAT-1 DC group, indicating a longer path taken by the T-cells when they are co-cultured with WT DCs (FIG. 6F). These data suggest that BMDCs derived from FAT-1 mice differentially express genes that may enhance their ability to stay in contact with a T-cell much longer.

[0193] Next, it was tested whether omega-3 fatty acids could improve the efficacy of WT-DCs by mimicking FAT- 1 -like phenotype.

[0194] Example 8: Omega-3 fatty acids and specialized pro-resolving lipid mediators (SPMs) partially mimic the FAT-1 phenotype

[0195] It was further tested whether a similar enhanced antigen presentation could be observed by generating DCs derived from BMs isolated from WT animals in the presence of omega-3 fatty acids. To do so, BM cells isolated from WT animals were cultured with various DHA / SPM doses (FIG. 7 A). Among the SPMs, RVD1, RvEl, RvE4, 17HDHA, PD1, and MR-1 were screened (FIG. 9). To test the antigen presentation ability of WT DCs generated in the presence of various omega-3 FAs, an antigen presentation assay was conducted by co-culturing PMEL-CD8 T cells with hgplOO pulsed mature WT DCs. The data indicated that hgplOO-pulsed mature DCs generated in the presence of DHA, RvDl, and RvE4 invoked a significantly higher IFNy response from the PMEL-CD8 T cells than those generated in the presence of the vehicle alone (FIGS. 7B, 7C, and 7E). A nonsignificant increase in IFNy production was observed in the case of RvEl (FIG. 7D). Together, this data suggests that elevated omega-3 levels during the generation of dendritic cells for cell-based vaccine therapy enhance the antigen presentation ability of the DCs, resulting in improved therapeutic efficacy, a finding that can be translated to human DC cancer vaccines.

[0196] Conclusions

[0197] The experiments described above demonstrate, for the first time, that a high level of omega-3 fatty acids can improve the antigen presentation ability of bone marrow-derived dendritic cells, thereby enhancing the therapeutic efficacy of DC-based cancer vaccines in murine tumor models. In addition to slower tumor growth, an increased ex vivo vaccine-specific response was found in tumor-bearing and naive WT mice when vaccinated with DCs with high endogenous omega-3 levels. Also reported herein is set of genes differentially expressed in DCs with high omega-3 compared to WT DCs. These data suggest that the omega-3-specific changes may enhance DC and CD8 T-cell interaction time (T-cell dwell time), resulting in better T-cell activation, IFNy production, and anti-tumor response.

[0198] Most studies have utilized omega-3 dietary supplementation to investigate their effects on physiology and diseases in rodents and humans. However, these prior studies were limited by animals' feeding behaviors and the uptake of fatty acids at the cellular level. The present studies used the FAT-1 mouse model, which has high endogenous levels of omega-3 fatty acids, to address this issue. FAT-1 mice express a C. elegans gene called FAT-1, which converts omega-6 fatty acids to omega-3 fatty acids by adding double bonds. As a result, these animals have high omega-3 levels and a low ratio of omega-6 to omega-3. Previous studies have reported that a high omega-6 / omega-3 ratio in the diet results in poor cardiovascular health and increased risk for obesity and cancer. Interestingly, FAT-1 mice exhibit lower body mass throughout their lifespan and significantly prolonged overall survival than WT mice. These findings demonstrate a direct effect of high omega-3 fatty acids on age-related weight gain and lifespan extension. In line with this observation, recent studies have reported a positive correlation between high omega-3 fatty acids and decreased mortality risk in humans.

[0199] FAT-1 mice demonstrated a decrease in tumor growth rate in various tumor models such as TC-1, B16F10, and EO771, similar to prior studies. However, it was also discovered that the survival rate of FAT- 1 mice with tumors was considerably longer than that of WT animals, which has not been previously reported. Studies indicate that cancer cells switch to the de novo fatty acid pathway to support rapid growth and metastasis and adapt mechanisms to eliminate omega-3 fatty acids from their surroundings. Therefore, the initial tumor growth in FAT-1 mice could be due to high omega-3 fatty acids' direct effect on tumor cells. However, the sustained slower growth and prolonged survival of tumor-bearing FAT-1 mice suggest a robust anti-tumor immune response. Thus, the present studies focused on the immunomodulatory effects of high omega-3, especially dendritic cell function, which is crucial for effectively priming T-cells.

[0200] Although no differences were observed in the frequencies of major immune cell subtypes at the basal level, a significant increase in the antigen presentation ability of bone marrow-derived dendritic cells from FAT-1 mice was observed in vitro. Additionally, there was a similar effect on antigen presentation when WT DCs were cultured in the presence of DHA. This observation is in stark contrast to previously published reports, which suggested an impaired antigen presentation due to a down-regulation of costimulatory molecules like CD80 and CD86 and reduced cytokine production, both in murine bone marrow- derived DCs and human monocyte-derived DCs in response to DHA treatment. There are several critical differences between the present study and the previously reported studies. First, a genetic mouse model was used instead of a dietary approach, which ensures a systemic increase in omega-3 levels. Second, the high omega-3 comes at the cost of omega-6, and the FAT-1 desaturase gene converts all omega-6 to omega-3. Third, the high omega-3 can remodel embryonic metabolism, leading to gene expression changes during fetal development, which cannot be achieved by the exogenous addition of omega-3 to already differentiated cells. In addition, the prior studies used a much higher concentration of DHA only during the maturation stage of DCs and did not study the effect of different doses on DC function. The concentration of DHA and other resolvins was optimized through careful titration. Moreover, the BMDCs were generated by culturing bone marrow cells in the continued presence of DHA, whereas the previously published studies only evaluated the effect of DHA during maturation. This critical distinction implicates an altered differentiation program of bone marrow cells when cultured constantly with DHA / resolvins, and suggests an approach to translate these discoveries into more effective human DC cancer vaccines.

[0201] The present disclosure reveals that antigen-pulsed BMDCs from FAT-1 mice can significantly reduce tumor growth and cachexia and increase overall survival in tumor-bearing WT mice. In addition, an enhanced vaccine-specific response was observed in both tumor-bearing and tumor-naive WT mice when vaccinated with antigen-pulsed BMDCs derived from FAT-1 mice. Increased PD-1+ CD8 T cells, a marker for T-cell activation and exhaustion, was also observed. This observation suggests that FAT-1 BMDCs effectively prime the T-cells, which can lead to exhaustion of the same by upregulation of PD-1. This presents an opportunity for combining anti-PD-1 checkpoint inhibitors with the FAT-1 BMDC vaccine to improve its efficacy further.

[0202] Also reported herein is the transcriptomic profile of bone marrow cells and immature and mature BMDCs from FAT-1 and wild-type mice. The bulk RNA sequencing from these cell types revealed differentially expressed genes that are explicitly upregulated in FAT-1 bone marrow cells, immature, and mature BMDCs. Several chemokines and cytokines, such as CXCL2, CCL2, CCL7, and CXCL7, with known immune cell recruitment functions, are upregulated in FAT-1 BMDCs. Interestingly, platelet factor 4 (PF4) / CXCL4 was among the top DEGs in DCs generated from FAT-1. Recent studies have reported that PF4 has a restorative effect on the aged brain, improving cognition by suppressing neuroinflammation and promoting synaptic plasticity. Furthermore, the study shows that this effect, in part, is due to the suppression of pro-aging immune factors and restoration of peripheral immunity to a more youthful phenotype by PF4. It is hypothesized that these PF-4-specific mechanisms may also play roles in FAT-1 mice, where a longer lifespan and robust anti -tumor immune response were observed.

[0203] Example 8: Production of mature DCs and DC vaccines from peripheral blood mononuclear cells

[0204] This example provides an exemplary protocol for producing mature, vaccine quality DCs from human peripheral blood mononuclear cells (PBMCs).

[0205] Preparation of media and buffers for DC vaccine manufacturing

[0206] Preparation of freeze media (FM): Freeze medium consists of 5% dimethyl sulfoxide (DMSO, an intracellular cryoprotectant), 6% pentastarch (hydroxyethyl starch, a volume expander and extracellular cryoprotectant), and PlasmaLyte A supplemented with 4% human serum albumin (HSA), heparin (anticoagulant, 30 U / ml) and Dornase Alfa (DNAse, 20 pg / ml, enzyme added to minimize clumping during product thaw). Cells are frozen at concentrations ranging from 106 / ml to 108 / ml depending on total volume available and planned usage.

[0207] Preparation of thaw medium (TM): Cells are typically thawed into the corresponding culture medium, usually 85% RPMI-1640, which can be supplemented with commercially available and 15% heat- inactivated pooled AB human serum (alternatively, heat-inactivated filtered (HIF) autologous or allogeneic AB plasma) supplemented with 10 lU / ml heparin.

[0208] Preparation of culture medium (CM): Culture medium is 90% RPMI-1640: 10% heat-inactivated pooled AB human serum (or HIF autologous or allogeneic AB plasma) supplemented with cytokines to encourage differentiation from monocyte to dendritic cell using 2000 lU / ml recombinant human (rh) IL-4 and 2000 lU / ml (rh) GM-CSF.

[0209] Preparation of dissociation buffer (DB): Cell Dissociation Buffer (Life Technologies, Grand Island, NY) is used to dissociate DCs adherent to the surface of culture containers and is enzyme-free, sterile and fully washed out from the final product prior to release. Preparation of infusion medium (IM): Cells are administered in a mixture of 10% heat-inactivated autologous plasma or 4% human serum albumin in Plasma-Lyte A.

[0210] Apheresis and monocyte elutriation

[0211] A subject undergoes large volume apheresis up to 25 liters to collect PBMCs and optionally autologous plasma (pooled human AB serum can replace autologous plasma) for DC vaccine manufacturing. The collected PBMCs undergo counter-flow centrifugation (elutriation) using a semi-closed system device such as a Terumo Elutra, or equivalent, for enrichment of autologous monocytes. Purity is expected to be >60% and recovery > 60%. If > 15% granulocyte contamination is observed, an additional manual density gradient separation using Ficoll-Paque Plus reagent can be performed. Enriched monocytes are aliquoted and cryopreserved in freezing medium (FM) with a target of up to 450xl06cells / vial for future preparation of dendritic cell products. Pooled AB serum is preferred, however, when autologous plasma is substituted then aliquots of up to 200 ml autologous plasma are retained per recovered cell vial and stored at -80°C.

[0212] DC vaccine manufacturing procedure

[0213] On Day 0, cells are thawed. If autologous plasma is being used, plasma is thawed in an over-wrap bag placed in 37 ± 1°C water bath. 25-50 ml of thaw medium (TM) is prepared per vial of cells. Enriched monocytes are retrieved from liquid nitrogen and placed in an over-wrap bag in 37 ± 1 °C water bath. When the vial is mostly liquid but still cold, cells are transferred to a 50 ml conical tube and TM is added very slowly to avoid rapid change in osmotic gradient that could lyse the cells. Centrifugation is performed at 1300 RPM (368 RCF) with deceleration setting 6 (DEC 6) for 10 minutes. The supernatant is removed and discarded. Next, the total culture volume (TCV) is determined by gently resuspending the cell pellet in 40 ml culture media (CM). Post-thaw specimen (1.0 ml) is transferred into a snap top tube to check cell count and viability. The target starting cell count per manufacturing is 120-350X105viable cells for a final product of 20xl06viable total nucleated cells (TNCs) on Day 4. TCV is calculated by using the formula viable TNC 1 1.5xl06 / ml (maximum allowed cell concentration per ml) and the type of the flask size and number is determined based on TCV. Maximum viable TNC to initiate culture at maximum allowed cell concentration of 1.5xl06 / ml is 350x10sat TCV in 233 ml. To initiate cell, culture, cells are diluted to the calculated TCV in the selected container(s) based on the total culture volume. The culture container(s) is placed in an incubator set at 37 °C 5% CO2.

[0214] No processing occurs on Day 1. On Day 2, cytokines in the culture are replenished by adding rh IL- 4 (2000 lU / ml), rh GM-CSF (2000 lU / ml) and keyhole limpet hemocyanin (KLH 10 mcg / ml, Stellar Biologies, Port Nueneme, CA) to CM to each culture container.

[0215] Day 3 differs depending on whether a peptide-pulsed DC vaccine or a viral vector transduced DC vaccine is being produced. For a peptide-pulsed DC vaccine, DCs are matured by adding medium containing IFN-y lb (1000 lU / ml, Horizon Therapeutics, Dublin, Ireland) and LPS endotoxin (15 ng / ml, List Biologies, Campbell, CA). A specimen (-24 hour timepoint, 2.5 ml) is removed and tested for sterility, and the culture flask is returned to the incubator set at 37°C 5% CO2. For a viral vector transduced DC vaccine, the viral vector needed for transduction is calculated at 3,000 viral particles per 1 mononuclear cell plated on the day of culture initiation, Day 0. Viral vector is thawed and added at the calculated volume to the culture flask. The flask is mixed gently by standing up and setting down the flask for 3-5 times and incubated for 4 hours at 37°C. Four hours after the initiation of transduction, IFN-y lb (1000 lU / ml) and LPS endotoxin (15 ng / ml, List Biologies, Campbell, CA) are added to the flask for DC maturation. The flask is mixed gently by standing up and setting down the flask for 3-5 times. A specimen is removed and tested for sterility, and the flask is returned to the incubator.

[0216] On Day 4, for peptide -pulsed DC vaccines, the product is collected for peptide pulsing. The culture container is rinsed twice with 50 ml pre-warmed HBSS. Rinses are transferred to 250 ml conical tube(s). 50- ml pre-warmed dissociation buffer (DB ) is pipetted into the culture container and placed in an incubator for 10 minutes. If cells do not dissociate from the culture container, the container is returned to the incubator. If >50% cells remain adherent, the DB step is repeated using pre-warmed DB and any removed culture media is saved. The culture container is rinsed with 30 ml HBBS by gentle swirling or rocking. The rinse with 50 ml HBBS is repeated. All rinsing media are collected in a 250 ml conical tube. The conical tubes that contain all media from rinsing are centrifuged at 1300 rpm (368 ref / xg) for 10 minutes at the deceleration setting 6 (DEC 6). Supernatants are removed from the conical tubes and 1 ml of the supernatant is transferred for mycoplasma testing along with other specimens. Cells are gently resuspended using a pipet (must not vortex). The cells are transferred to a 50 ml tube and cells are diluted to 42 ml with infusion media (IM) by gentle mixing. 0.5 ml is removed and checked for cell count and viability.

[0217] DCs are resuspended in pulsing volume of 10 ml using 15 ml conical tubes. Optimized concentration of peptide (in some instances, 5-20 Ltg / rnl) are added to each DC aliquot. DC and peptide mixes are individually transferred to cell culture bags and kept on a rocker (tilt 20°, speed 15 rpm) for 90 minutes in an 37°C incubator. Drops (4-5) of each peptide -pulsed DC preparation are collected in separate tubes by using 30 ml syringes. All remaining content in individual bags is transferred to one 250 ml conical tube and the tubes kept on a chilled metal block in an insulated container. If the cell suspension is visibly clumped, the suspension is filtered using 100 Ltrn cell strainer. The volume needed to package 20X106viable TNC is then calculated.

[0218] On Day 4, for viral transduced DC vaccines, cells are harvested and washed. Cell harvest can start 24 hours from the initiation of transduction. Two 250 ml conical tubes are placed on wet ice or in a chill block. Suspension cells are transferred to the first 250 ml conical tube. The flask is rinsed twice with 40 ml of cold HBSS per flask and retained in same 250 ml conical tube. Dissociation Buffer (DB) is added (40 ml per flask) and the flask is stored at 2-8°C for 30 minutes to detach adherent cells. The sides of the flask are tapped a few times and DB with cells is transferred to the second 250 ml conical tube, designated D4 harvest. The flask is rinsed with 40 ml cold HBSS and the rinse is transferred to the D4 conical tube. Both tubes are centrifuged, the supernatants are discarded, and the cell pellets are gently resuspended with HBSS (do not vortex). The contents of both harvest tubes are collected into one 50 ml conical and diluted to 50 ml with HBSS. The conical tube is centrifuged for 10 minutes at 1300 RPM (491 RCF) with deceleration setting (DEC 6) if a tabletop. The supernatant is discarded and the cells are gently resuspended to a final of 40 ml with IM. A sample is removed for sterility assays (e.g., STAT Gram stain, mycoplasma and endotoxin assay), flow cytometry, cell counts and viability. The volume needed to package the ordered cell dose is calculated and removed to a conical tube and centrifuged for 10 minutes using a tabletop centrifuge at 1300 RPM (491 RCF) with deceleration setting 6 (DEC 6). Following centrifugation, excess IM is removed and the cell pellet is gently resuspended in 0.5 ml of IM. Cells are retrieved cells with a 3 ml syringe or a pipette. The conical tube is rinsed with the remaining 0.5 ml of the IM to collect a final DC vaccine volume of 20X106cells in 1.0 ml.

[0219] It will be apparent that the precise details of the methods or compositions described may be varied or modified without departing from the spirit of the described aspects of the disclosure. We claim all such modifications and variations that fall within the scope and spirit of the claims below.

Claims

CLAIMS1. A method of producing a dendritic cell (DC)-based tumor vaccine, comprising: culturing bone marrow cells in culture media comprising granulocyte macrophage colony stimulating factor (GM-CSF) and an omega-3 lipid, thereby producing immature DCs; and(a) culturing the immature DCs with lipopolysaccharide (LPS) to produce mature DCs, and pulsing the mature DCs with a tumor antigen to produce mature DCs expressing the tumor antigen; or(b) transducing the immature DCs with a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen, and culturing the transduced immature DCs with LPS to produce mature DCs expressing the tumor antigen.

2. The method of claim 1, wherein the culture media comprises about 20 ng / ml GM-CSF.

3. The method of claim 1 or claim 2, wherein the immature DCs are cultured with about 10 ng / ml LPS.

4. The method of any one of claims 1 -3, wherein the mature DCs are pulsed with about 5 pg / ml to about 20 pg / ml of the tumor antigen.

5. The method of any one of claims 1-4, wherein the bone marrow cells are cultured with GM- CSF and the omega-3 lipid for about 7 days.

6. A method of producing a dendritic cell (DC)-based tumor vaccine, comprising: culturing monocytes in culture media comprising GM-CSF, interleukin-4 (IL-4) and an omega-3 lipid, thereby producing immature DCs; and(a) culturing the immature DCs with lipopolysaccharide (LPS) and interferon (IFN)-y lb to produce mature DCs, and pulsing the mature DCs with a tumor antigen to produce mature DCs expressing the tumor antigen; or(b) transducing the immature DCs with a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen, and culturing the transduced immature DCs with LPS and IFN-y lb to produce mature DCs expressing the tumor antigen.

7. The method of claim 6, wherein the monocytes are derived from a peripheral blood mononuclear cell sample.

8. The method of claim 6 or claim 7, wherein the culture media comprises about 2000 lU / ml ofGM-CSF.

9. The method of any one of claims 6-8, wherein the culture media comprises about 2000 lU / ml IL-4.

10. The method of any one of claims 6-9, wherein the culture media further comprises keyhole limpet hemocyanin (KLH).

11. The method of any one of claims 6- 10, wherein the culture media further comprises pooled AB serum, autologous plasma, or allogeneic AB plasma.

12. The method of any one of claims 6-12, wherein the immature DCs are cultured with about 15 ng / ml of LPS.

13. The method of any one of claims 6-13, wherein the immature DCs are cultured with about lOOO IU / ml of lFN-y lb.

14. The method of any one of claims 1-13, wherein the culture media is RPMI 1640 media.

15. The method of any one of claims 1-14, wherein the omega-3 lipid comprises docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a pro-resolvin lipid.

16. The method of claim 15, wherein the omega-3 lipid comprises DHA.

17. The method of claim 15, wherein the omega-3 lipid comprises EPA.

18. The method of claim 15, wherein the omega-3 lipid comprises a pro-resolvin lipid.

19. The method of claim 18, wherein the pro-resolvin lipid is selected from resolvin DI(RvDl), resolvin D2 (RvD2), resolvin El (RvEl), resolvin E4 (RvE4), protectin DI (PD-1), maresin 1 (MR- 1), maresin 2 (MR-2), and 17-hydroxy docosahexaenoic acid (17-HDHA).

20. The method of any one of claims 1-19, wherein the culture media comprises: about 0.1 pM to about 5 pM of the omega-3 lipid; about 0.3 LIM to about 4 pM of the omega-3 lipid; or about 0.5 pM to about 3 pM of the omega-3 lipid.

21. The method of claim 20, wherein the culture media comprises about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM, or about 3 pM of the omega-3 lipid.l ’ l. The method of claim 20 or claim 21, wherein the culture media comprises about 0.1 U.M to about 5 U.M of DHA.

23. A DC -based vaccine produced according to the method of any one of claims 1-22.

24. The DC -based vaccine of claim 23, wherein the DCs have an increased level of omega-3 lipid in their plasma membrane compared to DCs generated in the absence of omega-3 lipid.

25. The DC -based vaccine of claim 23 or claim 24, wherein the DCs exhibit increased T-cell dwell time compared to DCs generated in the absence of omega-3 lipid.

26. The DC -based vaccine of any one of claims 23-25, the DCs exhibit enhanced T-cell activation and / or IFNy production compared to DCs generated in the absence of omega-3 lipid.

27. A method of treating cancer in a subject, comprising administering to the subject a DCbased tumor vaccine produced according to the method of any one of claims 1-22, or administering to the subject the DC-based vaccine of any one of claims 23-26, thereby treating cancer in the subject.

28. The method of claim 27, further comprising administering to the subject a checkpoint inhibitor.

29. The method of claim 28, wherein the checkpoint inhibitor is a programmed cell death 1 (PD-1) inhibitor.

30. The method of any one of claims 27-29, wherein the cancer comprises a solid tumor.

31. The method of claim 30, wherein the solid tumor is a lung tumor or a melanoma.

32. The method of any one of claims 27-31, wherein: the size or number of tumors in the subject is reduced by at least 25%; cachexia is reduced by at least 25%; and / or survival of the subject is increased by at least 25%, as compared to a subject not receiving the DC -based vaccine or as compared to a subject receiving a DC -based vaccine, wherein the monocytes or bone marrow cells used to generate the DCs were not cultured with an omega-3 lipid.

33. The method of any one of claims 27-32, wherein the subject is a mammal.

34. The method of claim 33, wherein the mammal is a human.

35. Use of a DC -based vaccine produced according to the method of any one of claims 1-22, or the DC-based vaccine of any one of claims 23-26 for treating cancer in a subject.

36. A composition comprising: isolated dendritic cells (DCs), isolated monocytes, or isolated bone marrow cells; and an omega-3 lipid.

37. The composition of claim 36, wherein the omega-3 lipid comprises docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a pro-resolvin lipid.

38. The composition of claim 37, wherein the pro-resolvin lipid selected from resolvin DI (RvDl), resolvin D2 (RvD2), resolvin El (RvEl), resolvin E4 (RvE4), protectin DI (PD-1), maresin 1 (MR- 1), maresin 2 (MR-2), and 17-hydroxy docosahexaenoic acid (17-HDHA).

39. The composition of any one of claims 36-38, wherein the isolated DCs, monocytes or bone marrow cells comprise a recombinant nucleic acid molecule, vector or virus expressing a tumor antigen.

40. The composition of any one of claims 36-38, wherein the isolated DCs, monocytes or bone marrow cells have been pulsed with a tumor antigen.

41. The composition of any one of claims 36-40, wherein the composition comprises: about 0.1 LlM to about 5 LlM of the omega-3 lipid; about 0.3 LlM to about 4 LlM of the omega-3 lipid; or about 0.5 pM to about 3 pM of the omega-3 lipid.

42. The composition of any one of claims 36-41, wherein the composition comprises about 0.1 pM, about 0.3 pM, about 0.5 pM, about 1 pM, about 1.5 pM, about 2 pM, about 2.5 pM, or about 3 pM of the omega-3 lipid.

43. The composition of any one of claims 36-42, further comprising a pharmaceutically acceptable carrier, an adjuvant, AB allogeneic plasma, pooled AB serum, or any combination thereof.

44. The composition of any one of claims 36-43, wherein the isolated DCs, monocytes or bone marrow cells have an increased level of omega-3 lipid in their plasma membrane compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid.

45. The composition of any one of claims 36-44, wherein the DCs, monocytes or bone marrow cells exhibit increased T-cell dwell time compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid.

46. The composition of any one of claims 36-45, wherein the DCs, monocytes or bone marrow cells exhibit enhanced T-cell activation and / or IFNy production compared to DCs, monocytes or bone marrow cells cultured in the absence of omega-3 lipid.

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