WO2009097545A1 - Bioactive sutures for the treatment of cancer - Google Patents

Abstract

The present invention relates to compositions and method for the treatment of cancer using a suture or dressing providing a plurality of tumor cell antigens and/or an immunostimulatory agent to a subject in need of such treatment.

Description

DESCRIPTION

BIOACTIVE SUTURES FOR THE TREATMENT OF CANCER

BACKGROUND OF THE INVENTION

[0001] The present invention claims priority to U.S. Provisional Patent Application Serial No. 61/025,440, filed February 1, 2008, which is incorporated by reference herein in its entirety.

I FIELD OF THE INVENTION

[0002] The present invention relates to the fields of oncology, immunology and biology. More particularly, the invention relates to sutures comprising one or more of (a) a tumor cell lysate and/or (b) an immunostimulator.

II BACKGROUND

[0003] Cancer constitutes one of the greatest health threats in the world, responsible for over one-half million deaths each year in the U.S. alone. Unfortunately, current treatment methods for cancer, including radiation therapy, surgery, and chemotherapy, are known to have limited effectiveness. Additional methods of cancer therapy are needed.

[0004] Cancer immunotherapy involves recruitment of the host's immune system to fight cancer. The central concept relies on stimulating the patient's immune system to attack tumor cells. Normally, the immune system responds to invasion on the basis of discrimination between self and non-self, but many kinds of tumor cells are tolerated by the patient's immune system, at least in part due to the fact that cells are essentially the patient's own cells. However, many kinds of tumor cells display unusual antigens that are not normally present on that type of cell. These antigens make ideal candidate targets for the immune system.

[0005] Antibodies are one component of the adaptive immune response, recognizing and stimulating an immune response to foreign antigens. A number of immunotherapeutic approaches to the treatment of cancer involve the use of antibodies. In particular, monoclonal antibodies make it possible to raise antibodies against specific tumor target antigens. Herceptin is an antibody against ErbB2 and was one of the first generation of immunotherapeutic treatments for breast cancer. However, the number of appropriate targets, and the corresponding development of safe and effective antibody therapeutics, has so far been limited.

[0006] Other types of immunotherapy also exist. For example, cytokines, such as IL-2, play a key role in modulating the immune response, and have been used in conjunction with antibodies in order to generate a greater immune response.

[0007] Yet another form of immunotherapy involves tumor vaccines. A large number of these vaccines, which involve the administration of either tumor antigens or genetics sequences encoding such antigens, have been attempted. However, tumor antigen variation and lack of immunogenicity still hamper this approach. Thus, additional immunotherapies for the treatment of cancer are needed.

SUMMARY OF THE INVENTION

[0008] Embodiments of the invention are directed to compositions and methods of use that include, in certain aspects, a surgical suture comprising (a) a biocompatible filament and (b) an immunostimulatory agent. In certain aspects, the immunostimulatory agent is present throughout the width of one or more sections of the biocompatible filament, more specifically, at a substantially uniform concentration throughout the width of one or more section of the filament. The "width" refers to a distance along the long axis of the filament. A width can include 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1 μm, nm, or mm, including all values and ranges there between of the length of the filament. In certain embodiments the immunostimulatory agent is present throughout the length of the biocompatible filament that is associated with a wound or treatment area. In a further apsect, the suture comprises one or more tumor cell antigens.

[0009] In certain aspects, ovalbumin or another adjuvant can be incorporated into the biocompatible filament. In still further aspects, the biocompatible filament may include only an immunostimulatory agent, e.g., CpG. The biocompatible filament could be biodegradable, such as a biodegradable polymer, i.e., a polymer that is broken down or dissolves when in contact with a living organism.

[0010] As used herein the term "biocompatible" refers to the quality of not having toxic or injurious effects on biological systems that such a material or composition comes in contact. "Immunostimulatory" refers to the ability of a molecule to activate either the adaptive immune system or the innate immune system. As used herein, "activation" of either immune system includes the production of constituents of humoral and/or cellular immune responses that are reactive against the immunostimulatory molecule and/or antigens associated with the immunostimulatory molecule. An adjuvant is an agent, such as ovalbumin, that stimulates the immune system and increase the response to a vaccine or antigen, without having any specific antigenic effect in itself. Known adjuvants include polypeptides, oils, aluminum salts, and virosomes.

[0011] In accordance with the present invention, there is provided a method of treating or preventing cancer in a subject comprising one or more steps including (a) surgically resecting or otherwise removing all or part of a tumor or cancer cell mass, (b) contacting or administering to the subject, specifically, the resection site, a composition comprising a bioactive suture or dressing comprising an immunostimulatory agent and optionally a plurality of tumor cell antigens. The suture may be biodegradable, e.g., composed of a biodegradable polymer.

[0012] The bioactive suture may comprise silk, elastin, chitin, chitosan, poly(d-hydroxy acid), poly(anhydrides), and/or poly(orthoesters). More particularly, the bioactive suture may comprise polyethylene glycol, poly(lactic acid), poly(glycolic acid), copolymers of lactic and glycolic acid, copolymers of lactic and glycolic acid with polyethylene glycol, poly(ε- caprolactone), poly(3-hydroxybutyrate), poly(p-dioxanone), polypropylene fumarate, poly(orthoesters), polyol/diketene acetals addition polymers, poly(sebacic anhydride) (PSA), poly(carboxybiscarboxyphenoxyphenoxy hexone (PCPP) poly[bis (p-carboxypheonoxy) methane] (PCPM), copolymers of SA, CPP and CPM, poly(amino acids), poly(pseudo amino acids), polyphosphazenes, derivatives of poly[(dichloro)phosphazenes] and poly[(organo) phosphazenes], poly-hydroxybutyric acid, or S-caproic acid.

[0013] The immunostimulatory agent may comprise bacterial cell components, nucleic acids, and/or cytokines. In particular, bacterial cell wall components, LPS, bacterial DNA, viral RNA, CpG oligonucleotides, double-stranded RNA, β-glucan, zymosan™, IL-2, IL-6, IL-7, IL-15, IFN-γ, IFN-α and/or GM-CSF are contemplated. More specifically, the immunostimulatory agent is CpG oligonucleotide.

[0014] A plurality of tumor cell antigens may comprise a tumor cell lysate, for example, from a breast cancer cell, a head and neck cancer cell, a lung cancer cell, a stomach cancer cell, an esophageal cancer cell, a skin cancer cell, a colon cancer cell, an ovarian cancer cell, a prostate cancer cell, a testicular cancer cell, a uterine cancer cell, a cervical cancer cell, a pancreatic cancer cell, or a liver cancer cell. The tumor cell lysate may also be derived from a neuroblastoma, a WiIm' s tumor, a rhabdoid tumor, a sarcoma (osteogenic or non-osteogenic), a hepatoblastoma, a rhabdomyosarcoma, a lymphoma, or a leukemia. A tumor cell lysate may comprise a partially purified (i.e., various components of the lysate being isolated from other components of the lysate using various methods know in the art) portion of lysate, a fraction of tumor cell lysate. The subject may suffer from or be at risk of developing recurrent cancer, metastatic cancer, or multi-drug resistant cancer.

[0015] The method may further comprise administering to the subject a second cancer therapy. The second cancer therapy may be gene therapy, other immunotherapy, brachytherapy, chemotherapy, radiotherapy, toxin therapy, hormonal therapy, or surgery, including surgical resection of tumor.

[0016] The cancer to be treated by the method includes, but is not limited to, a neuroblastoma, a melanoma, a WiIm' s tumor, a rhabdoid tumor, a sarcoma (osteogenic or non-osteogenic), a hepatoblastoma, a rhabdomyosarcoma, a lymphoma, a leukemia, a breast cancer, a head and neck cancer, a lung cancer, a stomach cancer, an esophageal cancer, a skin cancer a colon cancer, an ovarian cancer, a prostate cancer, a testicular cancer, a uterine cancer, a cervical cancer, a pancreatic cancer, or a liver cancer.

[0017] A polymer may be polylactide-co-glycolide and the immunostimulatory agent is CpG oligonucleotide. The polymer may be polylactic acid and polyethylene glycol, and the immunostimulatory agent is CpG. These compositions may further comprise GM-CSF or other immunostimulatory agents.

[0018] In yet other embodiments, there is provided a kit comprising (a) a bioactive suture; (b) a plurality of tumor cell antigens; and (c) an immunostimulatory agent. The bioactive suture may comprise silk, elastin, chitin, chitosan, poly(d-hydroxy acid), poly( anhydrides), and poly(orthoesters). More particularly, the biocompatible polymer may comprise polyethylene glycol, poly(lactic acid), poly(glycolic acid), copolymers of lactic and glycolic acid, copolymers of lactic and glycolic acid with polyethylene glycol, poly(ε-caprolactone), poly(3-hydroxybutyrate), poly(p-dioxanone), polypropylene fumarate, poly(orthoesters), polyol/diketene acetals addition polymers, poly(sebacic anhydride) (PSA), poly(carboxybiscarboxyphenoxyphenoxy hexone (PCPP) poly[bis (p-carboxypheonoxy) methane] (PCPM), copolymers of SA, CPP and CPM, poly(amino acids), poly(pseudo amino acids), polyphosphazenes, derivatives of poly[(dichloro)phosphazenes] and poly[(organo) phosphazenes], poly-hydroxybutyric acid, or S-caproic acid. The immunostimulatory agent may comprise bacterial cell components, nucleic acids, and cytokines. Immunostimulatory agents include, but are not limited to LPS, bacterial DNA, viral RNA, CpG oligonucleotides, double-stranded RNA, β-glucan, zymosan™, IL-2, IL-6, IL-7, IL- 15, IFN-γ, IFN-α and GM- CSF.

[0019] The tumor cell antigens may comprise all or part of a tumor cell lysate, for example, a lysate of breast cancer cell, head and neck cancer cell, lung cancer cell, stomach cancer cell, esophageal cancer cell, skin cancer cell, colon cancer cell, ovarian cancer cell, prostate cancer cell, testicular cancer cell, uterine cancer cell, cervical cancer cell, pancreatic cancer cell, or liver cancer cell.

[0020] It is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein.

[0021] The use of the word "a" or "an" when used in conjunction with the term "comprising" in the claims and/or the specification may mean "one," but it is also consistent with the meaning of "one or more," "at least one," and "one or more than one."

[0022] These, and other, embodiments of the invention will be better appreciated and understood when considered in conjunction with the following description and the accompanying drawings. It should be understood, however, that the following description, while indicating various embodiments of the invention and numerous specific details thereof, is given by way of illustration and not of limitation. Many substitutions, modifications, additions and/or rearrangements may be made within the scope of the invention without departing from the spirit thereof, and the invention includes all such substitutions, modifications, additions and/or rearrangements.

BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein: [0024] FIG. 1 Schematic showing equipment and preparation of polymer strands or sutures loaded with CpG ODN and tumor lysate.

[0025] FIG. 2 Scanning electron microscopy image of biodegradable polymer strand or suture loaded with CpG ODN and tumor lysate

[0026] FIG. 3 Tumor lysate release profile from polymer strands or sutures

[0027] FIG. 4 CpG ODN release profile from polymer strands or sutures

[0028] FIG. 5 Graph showing size of neuroblastoma tumor growth after resection and placement or stitching of polymer strands

[0029] FIG. 6 Corresponding images of mice treated with polymeric strands loaded with CpG ODN and tumor lysate after resection of neuroblastoma.

[0030] FIGs. 7A-7D (FIGs. 7 A, 7B) Photograph of suture extrudate production. CpG ODN 1826 and ground PLGA pellets (75/25 0.47 dL/g) are placed into the hopper. At an operational temperature of 7O⁰C for the rotor, flexible suture material is formed. (FIG. 7C) Scanning electron micrographs show the initial solid structure and surface morphology of the CpG ODN loaded sutures (scale bar = 0.4 mm). (FIG. 7D) Release profiles of CpG ODN measured using UV spectroscopy at 260 ran show sustained release of CpG ODN for over a month (n=3 +/- SD). Images and release profiles are representative of at least two repeats.

[0031] FIGs. 8A-8D Tumor growth (FIG. 8A) and survival (FIG. 8B) of mice inoculated with 1 x 10⁶ wild-type tumor cells. Subcutaneous tumors were grown between 5 to 10 mm following which mice underwent resection of the local disease. The wound at the site of resection was closed and four groups of mice were compared for recurrent tumor growth and survival. Group 1 had wounds that were closed using commercial polyglycolic acid Vicryl suture (n=10), group 2 was injected with CpG ODN locally and wounds closed using Vicryl (n=10), group 3 had wounds closed using Vicryl and sutures loaded with CpG ODN were implanted into the same mice remotely on the opposite side of the resection site (n=10), and group 4, the wounds were closed with Vicryl and CpG ODN loaded suture (n=10). Growth curves in each group were compared to each other and to controls. (FIG. 8A) Tumor volume estimates from mixed linear models analysis of the 4 groups of mice. Tumor volume (cm³) is plotted as the mean +/- SD. Growth in group 4 was significantly different to all other groups (P<0.05). (FIG. 8B) Kaplan-Meier plot of the estimated survival functions for the 4 groups of mice. Survival in group 4 was significantly different to all other groups (P<0.05). Tumor growth (FIG. 8C) and survival (FIG. 8D) in mice depleted of CD4, CD8 T cells and NK cells. Subcutaneous tumors were grown between 5 to 10 mm following which mice underwent resection of the local disease. The site of resection was closed using Vicryl and the CpG ODN loaded sutures. Depletion of NK cells significantly increased tumor growth and reduced survival in comparison to all other groups (P<0.001). The data presented is representative of at least two sets of experiments.

DETAILED DESCRIPTION

[0032] Present therapeutic strategies for solid tumors include chemotherapy and radiation therapy following resection. A large portion of patients have not been cured by these therapies. As a result, alternative strategies, including immunotherapy, are being increasingly investigated. With the characterization of tumor antigens as targets for tumor-specific T cells, immunotherapy has once more become a major interest in the treatment of cancer. Ligands for Toll-like receptors (TLRs) have shown significant potential in stimulating strong immune responses against a variety of carcinomas. The inventors have recently shown that immunostimulators, such as cytosine-phosphorothioate-guanine oligodeoxynucleotide motifs (CpG ODN or CpG), that bind to intracellular TLR9 produce much stronger CD8+ responses when they are delivered in biodegradable microparticles. In certain aspects of the invention, bioactive, biodegradable sutures are used to close the site of resection and to provide sustained release of antigens and/or immunostimulator simultaneously to provide immunotherapeutic treatment. In addition, these immunostimulatory sutures can be packaged, scaled up, and stored easily.

[0033] As discussed above, there is a need for additional therapies for multiple diseases including cancer. Many cancerous tumors are ineffectively treated by standard treatment strategies such as surgery, chemotherapy, and radiation therapy. Immunotherapy, and specifically tumor vaccination, constitute an as of yet unrealized approach that has great potential due to its specificity and lack of toxicity. A primary concern is the inability to deliver the proper signals and antigens for vaccination when attempting to shape the appropriate immune response. Bioactive sutures offer a means of overcoming these limitations as they provide sustained release of antigen and/or immunostimulator at the tumor or tumor resection site. [0034] The inventors have developed bioactive sutures for tumor immunotherapy by loading them with a plurality of tumor cell antigens and/or immunostimulatory agents for induction of potent, effective immunity against the targeted tumor or minimal residual disease. Tumor cell antigen or tumor antigen is a substance produced in tumor cells that triggers an immune response in the host (e.g., a protein, peptide, polysaccharide, etc.). Tumor cell antigens are useful in identifying tumor cells and are potential candidates for use in cancer immunotherapy. In specific embodiments, tumor cell antigens comprise a tumor cell lysate or a fraction of tumor cell lysate. One benefit of this approach is the ability to load multiple antigens from a single autologous tumor or multiple tumors in the context of immunostimulatory agents for antigen presenting cells in a particular location.

[0035] One of the previous problems with inducing adequate immunity against tumors is the lack of adequate tumor antigens and the inefficient presentation of antigens to antigen presenting cells. However, aspects of the present invention overcome these shortcomings, as tumor lysate contains one or more tumor antigen epitopes. Moreover, in certain embodiments, there is a continual release of antigen/immunostimulatory agent from the sutures, which may sustain and further the immune response. In certain aspects, the inventors have found these sutures to be more effective than attenuated whole tumor cell or peptide vaccination in their ability to suppress established tumor growth and induce tumor-specific cellular immunity. Furthermore, coated devices with surface-bonded antigens do not provide sustained release of antigen.

I. BIOACTIVE COMPOSITIONS

[0036] Bioactive compositions of the invention include sutures and/or wound dressing, e.g., a therapeutic filament to be placed in contact with a region having or suspected of having cancer cells, or a region/site having a tumor resected or removed. The suture and/or wound dressing are used to deliver/release tumor antigens and/or immunostimulators for treating cancer, particularly, cancer recurrence following resection. In certain aspects, the suture is a biodegradable polymer.

[0037] The present invention affords the ability to manipulate various variables in the antigen/immunostimulator delivery process. In particular, the method of controlling the delivery/release rate can be tuned by controlling the characteristics of the polymer/filament or fiber composition, such as chemical composition, draw rate, mode of fabrication, and structural design. Methods of controlling release profile by chemical composition include: selection of different polymer types with different rates of biodegradability, use of polymers of differing molecular weights, incorporation of various additives to the polymer matrix, and use of different tumor antigens or immunostimulatory agents. The mode of fabrication may be adjusted to control the dimension of the suture/dressing, for example, the draw rate may be adjusted to vary the mean diameter of the suture extrudate - increasing the draw rate decreases the mean diameter of the suture extrudate.

A. Sutures

[0038] The term "suture" as used throughout the body of the present application is intended to comprise a variety of flexible securing elongated filaments whether they be made of natural filaments, synthetic or polymeric filaments, or other materials. Sutures may be introduced into the tissue preferably by a sharp metal needle attached to one end of the suture and are used to make "stitches" to close the wound for holding tissues together for healing and regrowth. Sutures can also be implanted at the site of tumor/tumor resections to act as a depot that provides local sustained release of biolmolecules. Generally, the suture needle is caused to penetrate and pass through the tissue pulling the suture through the tissue. The opposing faces of the tissue are then moved together, the needle is removed, and the ends of the suture are tied in a knot, therefore the suture forms a loop as the knot is tied. Sutures could be strong (so they do not break), non-toxic and hypoallergenic (to avoid adverse reactions in the body), and flexible (so they can be tied and knotted easily). In addition, they may lack the so called "wick effect," which means that sutures do not allow fluids to penetrate the body through them from the outside, which could easily cause infections.

[0039] Sutures are divided into two kinds - those that are absorbable or biodegradable and will break down harmlessly in the body over time without intervention, and those that are non-absorbable or non-biodegradable and may be manually removed if they are not left indefinitely. The type of suture used varies on the operation, with a major criteria being the demands of the location and environment. Preferably, biodegradable sutures are used in the present invention for controlled release of immunostimulatory agents.

[0040] Non-absorbable internal sutures require re-opening for removal. Sutures which lie on the exterior of the body can be removed within minutes, and without re-opening the wound. As a result, absorbable sutures are often used internally; non-absorbable externally.

[0041] Suture sizes are defined by the United States Pharmacopeia (U.S.P.). Sutures were originally manufactured ranging in size from #1 to #6, with #1 being the smallest. A #4 suture would be roughly the diameter of a tennis racquet string. The manufacturing techniques did not allow thinner diameters. As the procedures improved, #0 was added to the suture diameters, and later, thinner and thinner threads were manufactured, which were identified as #00 (#2-0 or #2/0) to #000000 (#6-0 or #6/0).

[0042] Typically, sutures range from #5 (heavy braided suture for orthopedics) to #11-0 (fine monofilament suture for ophthalmics). Atraumatic needles are manufactured in all shapes for most sizes. The actual diameter of thread for a given U.S.P. size differs depending on the suture material class.

[0043] Sutures to be placed in a stressful environment, for example the heart (constant pressure and movement) or the bladder (adverse chemical presence) may require specialized or stronger materials to perform their role; usually such sutures are either specially treated, or made of special materials, and are often non-absorbable to reduce the risk of degradation.

[0044] In accordance with the present invention, bioactive sutures may be used to deliver tumor antigens and/or immunostimulatory agents of the present invention. In some embodiments, bioactive sutures comprise biocompatible filaments, which, for example, comprise biodegradable polymers, where "biocompatible" means there is no toxic, injurous, or adverse effects on biological systems. A variety of polymer-based sutures can be employed in this context.

[0045] The term "biodegradable" is synonymous with "bioabsorbable" and is art- recognized. It includes polymers, compositions, and formulations, such as those described herein, that degrade during use, particularly in contact with a living organism. In general, degradation attributable to biodegradability involves the degradation of a biodegradable polymer into its component subunits, or digestion, e.g., by a biochemical process, of the polymer into smaller, non-polymeric subunits. In certain embodiments, biodegradation may occur by enzymatic mediation, degradation in the presence of water and/or other chemical species in the body, or both.

[0046] The degradation rate of a biodegradable polymer often depends in part on a variety of factors, including the chemical identity of the linkage responsible for any degradation, the molecular weight, crystallinity, biostability, and degree of cross-linking of such polymer, the physical characteristics (e.g., shape and size) of the implant, and the mode and location of administration. For example, the greater the molecular weight, the higher the degree of crystallinity, and/or the greater the biostability, the biodegradation of any biodegradable polymer is usually slower.

[0047] In certain embodiments wherein the biodegradable polymer also has a therapeutic agent or other material associated with it, the biodegradation rate of such polymer may be characterized by a release rate of such materials. In such circumstances, the biodegradation rate may depend not only on the chemical identity and physical characteristics of the polymer, but also on the identity of material(s) incorporated therein.

[0048] In certain embodiments, polymeric formulations of the present invention biodegrade within a period that is acceptable in the desired application, hi certain embodiments, such as in vivo therapy, such degradation occurs in a period usually less than about five years, one year, six months, three months, one month, fifteen days, five days, three days, or even one day on exposure to a physiological solution with a pH between 6 and 8 having a temperature of between 25 and 37°C. In other embodiments, the polymer degrades in a period of between about one hour and several weeks, depending on the desired application.

[0049] The biocompatible polymer may comprise silk, elastin, chitin, chitosan, poly(d- hydroxy acid), poly(anhydrides), and poly(orththoesters). More particularly, the biodegradable polymer may comprise polyethylene glycol, poly(lactic acid), poly(glycolic acid), copolymers of lactic and glycolic acid, copolymers of lactic and glycolic acid with polyethylene glycol, poly(ε-caprolactone), poly(3-hydroxybutyrate), poly(p-dioxanone), polypropylene fumarate, poly(orthoesters), polyol/diketene acetals addition polymers, poly(sebacic anhydride) (PSA), poly(carboxybiscarboxyphenoxyphenoxy hexone (PCPP), poly[bis (p-carboxypheonoxy) methane] (PCPM), copolymers of SA, CPP and CPM, poly(amino acids), poly(pseudo amino acids), polyphosphazenes, derivatives of poly[(dichloro)phosphazenes] and poly[(organo) phosphazenes], poly-hydroxybutyric acid, or S-caproic acid.

[0050] Polylactide-co glycolide (PLGA), a biodegradable polymer, can serve as a structural matrix in which therapeutic agents are incorporated in the long-term delivery systems. The final products of PLGA degradation are lactic acid and glycolic acid, which are water soluble, non-toxic products of normal metabolism. See also U.S. Patents 6,884,435; 5,603,960; and 6,913,767. [0051] Polylactic acid (poly-lactide; PLA) is a polymer known for its ability to biodegrade. Since it does biodegrade, and can be processed to have such a wide variety of properties, it can be used in surgical sutures.

[0052] Polyethylene glycol (PEG) is a water-soluble, waxy solid that is used extensively in the cosmetic and toiletry industry. As the molecular weight of PEG increases, viscosity and freezing point increase. Although PEG is water soluble, solubility is greatly reduced at temperatures approaching 0°C, allowing experiments to run for 15-20 minutes before dissolution of PEG becomes pronounced. At higher temperatures (above 10°C) this length of time is much shorter.

B. Wound Dressing

[0053] Wound dressing is a therapeutic or protective material applied to a wound or postsurgical structures. Wound dressing may be made from biodegradable polymer as described above. In certain aspects, compositions of the invention include wound dressing such as patches and the like (described below) comprising a plurality of tumor cell antigens and an immunostimulatory agent. Patches are small pieces of material used to mend a wound or cover a resection bed. In medicine, surgical patches are pieces of synthetic material or biological tissue used to bridge together the defect between the edge of an incision or a gap in a biological structure. Patches are also used after certain surgeries to strengthen the repaired area. A variety of synthetic patches are available from medical device companies such as IMPRA, WL Gore, Sulzer Vascutek, Shelhigh, Bio Nova International, Intervascular and Aesculap for example.

[0054] Soft silicone-containing dressing is a type of dressing that uses the man-made material silicone in its adhesive, as well as in its wound contact layer. Silicone helps prevent the dressing from sticking to the wound or to the surrounding skin. This results in less trauma to the area as the dressing is repositioned or removed, and therefore aids in the healing process. Dressings incorporating soft silicone are designed for wounds with a wide range of drainage.

[0055] Hydrocolloid dressings are formulations of elastic, adhesive, and gelling agents (such as pectin or gelatin) and other absorbent ingredients. When applied to a wound, the wound drainage interacts with the dressing's components to form a gel-like substance that provides a moist environment for wound healing. Hydrocolloid dressings come in several shapes, sizes, and thicknesses, and are used on wounds with light to medium levels of wound drainage. This type of dressing is typically changed once every five to seven days, depending on the method of application, location of the wound, degree of exposure to "friction and shear," and incontinence. Hydrocolloid dressings are not usually used on wounds that have become infected.

[0056] Hydrogels are available in sheets, saturated gauze, or a gel. Gels provide a soothing and cooling effect on the wound, which promotes patient comfort. Gels are excellent for creating or maintaining a moist healing environment and are used on wounds with low levels of wound drainage. Gels are applied directly to the wound, do not adhere to the wound, and are usually covered with a secondary dressing to maintain the moisture level needed to promote wound healing.

[0057] Hydrofibers are soft non-woven pad or ribbon dressings made from sodium carboxymethylcellulose fibers, the same absorbent material used in hydrocolloid dressings. The dressing components interact with wound drainage to form a soft gel that can be easily removed. Hydrofibers are used on wounds with a heavy level of wound drainage, and wounds that are deep and need packing. Hydrofibers can also be used on a dry wound as long as the dressing is kept moist (by adding normal saline solution). This type of dressing can also control minor bleeding. Hydrofibers require a cover dressing. Hydrofiber dressings may stay in place for up to seven days depending on the amount of drainage from the wound.

[0058] Alginates are soft, non-woven fibers derived from brown seaweeds, particularly the kelps. Alginates are available in pad or rope form. Alginates and hydrofibers are similar types of products. In this case, the alginate product itself turns into a soft, non-adhesive gel when in contact with wound drainage. Alginates are used on wounds with a moderate to heavy level of wound drainage and can also control minor bleeding. Alginates require a cover dressing and should not be used in dry wounds. Dressings may be cut to fit the size of the wound and loosely packed, or might be layered for additional absorbency.

[0059] Gauze dressings are made from woven and non-woven fibers of cotton, rayon, polyester, or a combination of these fibers. Most woven products are a fine or coarse cotton mesh, depending on the thread count per inch. Fine mesh cotton gauze is frequently used for packing, such as normal saline wet-to-moist dressings. Coarse mesh cotton gauze, such as a normal saline wet-to-dry dressing, is used for non-selective debriding (the removal of dead/dying tissue and debris). Most non- woven gauze dressings are made of polyester, rayon, or blends of these fibers and appear to be woven like cotton gauze but are stronger, bulkier, softer, and more absorbent. Some dressings, such as dry hypertonic saline gauze used for debridement, contain substances to promote healing. Other products contain petrolatum or other wound healing elements indicated for specific types of wounds.

II. TUMOR CELL LYSATES AND ANTIGENS

[0060] In accordance with certain aspects of the present invention, there is provided a suture or wound dressing comprising one or more tumor antigen and/or a tissue lysate derived from cancer cells, cancerous tissue, tumor, or tumor sample. In one embodiment, the source of tissue lysates is the patient to be treated. In another embodiment, the tissue lysate can come from a bank of similar tumors from multiple patients. Generally, standard biopsy procedures can be used to obtain samples from solid tumors that can then be lysed to produce tumor lysates. Biopsy procedures will generally involve sterility. In various embodiments, sterility will be maintained even if the tissues being sampled are from cadavers or animals that will be sacrificed. To harvest the tissues of the invention, biopsies can be performed percutaneously with or without radiologic guidance or via incisions that will be made proximal to the tissue of interest, followed by retraction, excision of tissue and surgical closing of the incision. Superficial tissue sites are accessed by simple excision of the available tissue. Appropriate physiologic buffers are generally applied to the tissue, or the tissues are immersed therein. The tissue may also be cooled to appropriate temperatures for limited periods of time. Steps should be taken to ensure that apoptosis or other cellular degradation will not be induced in the tissue specimen.

[0061] If needed, the tissue samples can be lysed using a variety of methods known in the art, such as physical disruption and detergent lysis. Physical methods employed to disrupt cells and/or tissues include freeze-thawing, sonication, shearing, irradiation or exposure to microwaves. A variety of detergents may be used to solubilize cells and/or tissues, including anionic, cationic, zwitterionic and non-ionic detergents. By virtue of their amphipathic nature, detergents are able to disrupt bipolar membranes. In selecting a detergent, consideration will be given to the nature of the target antigen(s), and the fact that anionic and cationic detergents are likely to have a greater effect on protein structure than zwitterionic or non-ionic detergents. In some embodiments, zwitterionic detergents offer efficient disruption of protein aggregation. Exemplary anionic detergents include chenodeoxycholic acid, N- lauroylsarconsine sodium salt, lithium dodecyl sulfate, 1-octanesulfonic acid sodium salt, sodium cholate hydrate, sodium deoxycholate, sodium dodecyl sulfate and glycodeoxycholic acid sodium salt. Cationic detergents include cetylpyridinium chloride monohydrate and hexadecyltrimethylammonium bromide. Zwitterionic detergents include CHAPS, CHAPSO, SB3-10 and SB3-12. Non-ionic detergents may be selected from N-decanoyl-N- methylglucamine, digitonin, n-dodecyl β-D-maltoside, octyl α-D-glucopyranoside, Triton X- 100, Triton X-114, Tween 20 and Tween 80.

[0062] Commercial sources of tumor lysates also are available. For example, Protein Biotechnologies (at World Wide Web address 'proteinbiotechnologies.com') sells lung, breast, colon, uterine, cervical, ovarian, and stomach tumor lysates.

III. IMMUNOSTIMULATORY AGENTS

[0063] A variety of agents may be incorporated into the suture or dressing compositions of the present invention as immunostimulatory agents. These generally fall into the categories of bacterial cell products, nucleic acids, cytokines and growth factors, and miscellaneous agents.

A. Bacterial Cell Products

[0064] Bacterial cell wall components such as lipopolysaccharide (LPS), peptidoglycan (PG), lipoteichoic acid (LTA), and lipopeptides/proteins (LP) represent such bacterial compounds present in Gram-negative and/or Gram-positive bacteria. They are able to activate cells of the innate and adaptive immune system, but also react with further cells like vascular cells and epithelial cells. Bacterial DNA also is able to stimulate immune response, as discussed below.

B. Nucleic Acids

[0065] Unmethylated CpG motifs are prevalent in bacterial but are rare in vertebrate genomes. Oligodeoxynucleotides containing CpG motifs activate host defense mechanisms leading to innate and acquired immune responses. The recognition of CpG motifs typically require Toll-like receptor (TLR) 9. Cells that express TLR-9, which include plasmacytoid dendritic cells (PDCs) and B cells, produce proinflammatory cytokines, interferons, and chemokines. CpG-driven innate immunity protects against challenge with a wide variety of antigens, including pathogens, allergens and cancer cells. Thus, CpG ODNs enhance the development of acquired immune responses. See also U.S. Patents 6,821,957, 6,653,292, 6,429,199, 6,406,705, 6,339,068, 6,239,116, 6,214,806, 6,207,646 and 6,194,388, each of which is incorporated herein by reference. [0066] Other nucleic acids that have immunostimulatory properties include bacterial DNA, viral RNA, and double-stranded RNA. Bacterial DNA is immunostimulatory largely due to unmethylated CpG motifs.

C. Cytokines

[0067] A variety of cytokines, interferons and other factors can be used to enhance the immune response to tumor antigens of the present invention. For example, granulocyte- macrophage colony-stimulating factor (GM-CSF) is a colony-stimulating factor that stimulates the production of white blood cells, especially granulocytes and macrophages, and cells (in the bone marrow) that are precursors of platelets. It is a cytokine that belongs to the family of drugs called hematopoietic (blood-forming) agents. It is also referred to as sargramostim.

[0068] Other cytokines that may be used in accordance with the present invention include, but is not limted to IL-2, IL-6, IL-7, IL-15, IFN-γ, and IFN-α.

D. Miscellaneous Agents

[0069] A variety of other immunostimulatory agents also may be used in accordance with the present invention. As an example, β-glucans are polysaccharides generally come from cultured extract of Baker's yeast cell wall. They are found bound together as a sugar/protein complex. Certain plants and microorganisms are naturally high in these polysaccharides. The richest concentrated source is Baker's yeast cell walls, but it also is present in lesser amounts in mushroom extracts and lentinen, barley, oat, etc. Sodium alginate is also an excellent source, but the high sodium content is a major drawback in the processing for supplemental use.

[0070] Another form of β-glucan is a research extract called Zymosan™ (Biosynth). It is mannan-rich and prepared according to Pillemer et al. (1956). Zymosan™ activates the alternative complement cascade. It becomes coated with C3b/C3bi and is therefore a convenient opsonized particle. It also leads to C5a - production in serum, it is a potent stimulator of alveolar macrophages and induces the release of cytokines, e.g., interleukin 8 (IL-8) from human neutrophils and proinflammatory cytokines in immune cells. The toll-like receptor 2 has been shown to be involved in Zymosan™ induced signaling. Zymosan™ also induces protein phosphorylation and inositol phosphate formation. IV. PREPARING POLYMER-ANTIGEN/IMMUNOSTIMULATOR COMPLEXES

[0071] In certain aspects of the present invention, a bioactive suture is prepared by encapsulating a therapeutic agent into a suture or wound dressing composition, such as a polymeric composition. In certain embodiments, "encapsulate" includes incorporating, formulating, or otherwise including such agent into a composition that allows for release, such as sustained release, of such agent in the desired application. The term contemplates any manner by which a therapeutic agent or other material is incorporated into a polymer matrix, including for example: attached to a monomer of such polymer (by covalent, ionic, or other binding interaction), physical admixture, enveloping the agent in a coating layer of polymer, and having such monomer be part of the polymerization to give a polymeric formulation, distributed throughout the polymeric matrix, appended to the surface of the polymeric matrix (by covalent or other binding interactions), encapsulated inside the polymeric matrix, etc. The therapeutic agent may be tumor antigen or immunostimulator as described above.

[0072] More specifically, the physical form in which any therapeutic agent or other material is encapsulated in polymers may vary with the particular embodiment. For example, a therapeutic agent or other material may be first encapsulated in a microsphere and then combined with the polymer in such a way that at least a portion of the microsphere structure is maintained. Alternatively, a therapeutic agent or other material may be sufficiently immiscible in the polymer of the invention that it is dispersed as small droplets, rather than being dissolved, in the polymer. Any form of encapsulation or incorporation is contemplated by the present invention, in so much as the release, preferably sustained release, of any encapsulated therapeutic agent or other material determines whether the form of encapsulation is sufficiently acceptable for any particular use.

[0073] Sutures can be prepared, for example, by mixing ground polymer, dye, immunostimulatory agent, and/or tumor antigens in a hopper, and extruding suture filaments. In certain aspect the polymer is PLGA and the immunostimulatory agent is CpG.

[0074] In one embodiment, sutures are prepared by loading a mixture of powdered PLGA polymer, dye, CpG ODN and lyophilized tumor lysate into a hopper of a polymer extruder. The hopper is initially loaded with PLGA powder alone, followed by the mixture with the dye. The mixture is slightly heated before being forced through an extruder. In this way, the sutures being used have CpG ODN and/or tumor lysate within the polymer matrix. [0075] One example of suture impregnation is provided in U.S. Patent publication 2003/008242, which is incorporated herein by references, hi general, a suture, e.g. a polymer, or a wound dressing material is immersed in a solution comprising immunostimulatory agents, tumor antigens and a dye, followed by air-drying and washing off excessive composition. The adhesive potential of the dye makes them self-adhesive to sutures.

[0076] Another example of suture manufacture is described in U.S. Patent 5,456,696, which is incorporated herein by references. In general, the conditions of the individual steps of extruding, drawing, and annealing in the monofilament suture manufacturing process can be substantially the same as those disclosed in U.S. Patent 5,217,485, the contents of which are hereby incorporated by reference. Similarly, the process herein can employ much the same type apparatus as that described in U.S. Patent 5,217,485.

[0077] A typical manufacturing operation for the extrusion, quenching, aging, and stretching operations of a monofilament are described here as an example. An extruder unit is of a known or conventional type and is equipped with controls for regulating the temperature of a barrel in and various zones thereof. Pellets or powder of polymer are introduced to the extruder through a drier-hopper. Suitable polymers include those that are bioabsorbable and nonbioabsorbable, as long as they have an appropriate release profile for antigens and immunostimulatory agents. Examples of bioabsorbable polymers which can be employed in the process include polymers, copolymers and polymeric blends derived from monomers known to provide biocompatible, bioabsorbable polymers. Such monomers include glycolide, glycolic acid, lactide, lactic acid, p-dioxanone, trimethylene carbonate, epsilon-caprolactone, dimethyltrimethylene carbonate, 1,5 dioxepen-2-one and the like. Examples of nonbioabsorbable polymers which can be employed in the process of this invention include polyethylene, polypropylene, nylon, polyethylene terephthalate, and the like.

[0078] Typically, a motor-driven metering pump delivers melted extruded polymer at a constant rate to a spin pack and thereafter through a spinneret possessing one or more orifices of desired diameter to provide a molten monofilament which then enters quench bath, e.g., containing water, where the monofilament solidifies. The distance the monofilament travels after emerging from spinneret to the point where it enters quench bath is the air gap and can vary from about 0.5 to about 100 cm. If desired, a chimney or shield, can be provided to reduce the length of the air gap, e.g., from 1 to 10 cm, thereby isolating monofilament from contact with air currents which might otherwise affect the cooling of the monofilament in an unpredictable manner. The monofilament is typically passed through the quench bath around a driven roller and over idle rollers. Optionally, a wiper may remove excess water from the monofilament as it is removed from the quench bath. On exiting the quench bath the monofilament can enter a first godet station.

[0079] A first godet station may be equipped with up to, at least, or at most five individual godets around which the monofilament is wrapped. The first godet may be provided with a nip roll to prevent slippage which might otherwise result. Upon entering the first godet station, the monofilament can pass over the first godet, under a second godet, over a third godet, under a fourth godet and over a fifth godet. Typically the fifth godet is proximally located to a separation roller that is provided with a plurality of laterally spaced circumferential grooves which can act as guides for the monofilament. After the monofilament passes over a fifth godet it wraps around a groove on the separation roller and extends back to and around a corresponding groove on a separation roller located proximal to the first godet. The monofilament can wrap around a separation roller, ascends up to the first godet and continues onward to the remaining godets in the manner just described. When the monofilament passes over the fifth godet a second time, it may be wrapped around a second groove on a separation roller. The monofilament then extends back to a separation roller and around a corresponding groove thereon. The monofilament may pass through the first godet station any desired number of times. The solidified monofilament is thus allowed to dwell at ambient conditions before the monofilament enters a heating unit. In this fashion the monofilament is aged or exposed to ambient conditions for a desired period of time prior to being stretched. The solidified monofilament can be exposed to ambient conditions for a predetermined extended period of time of at least two minutes.

[0080] Aging or exposing the monofilament to ambient conditions for a predetermined period of time prior to drawing the monofilament can be accomplished in many different ways. For example, any number of godets may be employed to provide the dwell period. In addition, the arrangement of the godets can be varied. Also, other structure suitable for providing aging of the monofilament prior to stretching will be apparent to those skilled in the art.

[0081] A monofilament passing from a godet station is stretched to effect its orientation and thereby further increase its tensile strength. Stretching may be achieved by cold drawing the monofilament or drawing the monofilament while or after it has been heated. In the stretching operation a monofilament is drawn through a heating unit by means of a second godet station which rotates at a higher speed than the first godet station to provide the desired stretch ratio. For larger size sutures, e.g., sizes 2 to 3/0, a heating unit may comprise a hot liquid bath (such as glycerol or water), through which a monofilament passes. For smaller size sutures, e.g., sizes 4/0 to 8/0, the heating unit may comprise a hot air convection oven chamber.

[0082] Following the stretching operation, a monofilament may be subjected to additional stretching operations or an on-line annealing (relaxation) operation as a result of which the monofilament may undergo shrinkage. In accordance with methods that are known and described in the prior art, on line annealing with or without relaxation when desired is accomplished by driving the monofilament station through a second heating unit by a third godet station. For relaxation, the third godet station rotates at a slower speed than the second godet station thus relieving tension on the filament.

[0083] Other methods of producing a suture material are also well known to the skilled artisan.

V. METHODS OF THERAPY

[0084] The present invention also involves the treatment of cancer. The types of cancer that may be treated are not limited other than that they be responsive to immunotherapy according to the present invention. Thus, it is contemplated that a wide variety of tumors may be treated using the immunotherapy of the present invention, including cancers of the brain, lung, liver, spleen, kidney, lymph node, pancreas, small intestine, colon, stomach, breast, endometrium, prostate, testicle, ovary, skin, head and neck, esophagus, or other tissue. In certain aspects the cancer is a tumor or a resectable tumor. Tumor resection refers to the removal of part of an effected (contains or is suspected of containing cancerous cells or tumors) organ or tissue.

[0085] In many contexts, it is not necessary that the tumor cell be killed or induced to undergo normal cell death or "apoptosis." Rather, to accomplish a meaningful treatment, all that is required is that the tumor growth be slowed to some degree. It may be that the tumor growth is completely blocked or that some tumor regression is achieved. Clinical terminology such as "remission" and "reduction of tumor" burden also are contemplated given their normal usage.

[0086] Administration of these compositions according to the present invention will typically be via suturing a target tissue, organ, or resected area via surgical methods. Of particular interest is direct intratumoral administration or administration local or regional to a rumor, for example, in a resected tumor bed.

A. Tumor Resection

[0087] Typically, compositions of the invention will be used in conjunction with surgical therapy that includes administering bioactive sutures at a site containing, suspected of containing, or in a location associated with a tumor or cancer cell localization (e.g., lymph nodes, resection bed, etc.) hi certain embodiments of the invention, methods include subjecting the patient to radiotherapy and/or chemotherapy prior to or after tumor resection/immunotherapy. In other particular embodiments, methods also involve resecting all or part of a tumor from the patient. It is contemplated that multiple tumors may be removed (whole or part). In each of these cases, a bioactive suture can be provided, before, during or after the other cancer therapy. In certain embodiments, a standard cancer therapy is provided to the patient after tumor resection, such as by administering a composition with one or multiple agents to at least the resulting tumor bed. Some possible surgical options for surgical treatment of cancer are a segmental or wedge resection, a lobectomy, or a pneumonectomy (e.g., removal of all or part of an organ or tissue).

B. Combined Therapy with Chemotherapy or Radiotherapy

[0088] Tumor cell resistance to traditional therapies represents a major problem in clinical oncology. One goal of current cancer research is to find ways to improve the efficacy of chemo- and radiotherapy. One way is by combining such traditional therapies with a new therapy. For example, the herpes simplex-thymidine kinase (HS-t&) gene when delivered to brain tumors by a retroviral vector system successfully induced susceptibility to the antiviral agent ganciclovir (Culver et al, 1992). In the context of the present invention, it is contemplated that the immunotherapy could be used similarly in conjunction with chemo- or radiotherapeutic intervention in combination with tumor resection. It also may prove effective to combine immunotherapy of the present invention with chemotherapy and/or radiotherapy, as described below.

[0089] To kill cells, inhibit cell growth, inhibit metastasis, inhibit angiogenesis or otherwise reverse or reduce the malignant phenotype of tumor cells using the methods and compositions of the present invention one would generally administer the immunotherapeutic composition of the present invention and at least one other agent. These compositions would be provided in a combined amount effective to kill or inhibit proliferation of the cell. This process may involve administering the immunotherapeutic composition and the other agent(s) or factor(s) at the same or different time. This may be achieved by administering two distinct compositions or formulations, at the same time, wherein one composition includes the immunotherapeutic compositions and the other includes the other agent.

[0090] Alternatively, the immunotherapy treatment (i.e., tumor resection and suturing with bioactive sutures) may precede or follow the other agent treatment by intervals ranging from minutes to weeks. In embodiments where the other agent and immunotherapeutic composition are applied separately, one would generally ensure that a significant period of time did not expire between the time of each delivery, such that the agent and the immunotherapy composition would still be able to exert an advantageously combined effect. In such instances, it is contemplated that one would contact the cell with both modalities within about 12-24 hours of each other and, more preferably, within about 6-12 hours of each other, with a delay time of only about 12 hours being most preferred. In some situations, it may be desirable to extend the time period for treatment significantly, however, where several days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7 or 8) lapse between the respective administrations.

[0091] It also is conceivable that more than one administration of the other agent will be desired. Various combinations may be employed, where resection/immunotherapeutic composition is "A" and the other agent is "B," as exemplified below:

[0092] B/A/B B/B/A A/B/B B/B/B/A B/B/A/B

[0093] B/B/B/A A/B/B/B B/A/B/B B/B/A/B

[0094] Other combinations are contemplated. Again, to achieve cell killing, both agents are delivered to a cell in a combined amount effective to kill the cell.

[0095] Agents or factors suitable for use in a combined therapy are any chemical compound or treatment method that induces DNA damage when applied to a cell. Such agents and factors include radiation and waves that induce DNA damage such as, γ-irradiation, X-rays, UV-irradiation, microwaves, electronic emissions, and the like. A variety of chemical compounds, also described as "chemotherapeutic agents," function to induce DNA damage, all of which are intended to be of use in the combined treatment methods disclosed herein. Chemotherapeutic agents contemplated to be of use, include, e.g., adriamycin, 5-fluorouracil (5FU), etoposide (VP- 16), camptothecin, actinomycin-D, mitomycin C, cisplatin (CDDP) and even hydrogen peroxide. The invention also encompasses the use of a combination of one or more DNA damaging agents, whether radiation-based or actual compounds, such as the use of X-rays with cisplatin or the use of cisplatin with etoposide.

[0096] In treating cancer according to the invention, one would contact the tumor cells with an agent in addition to the immunotherapeutic composition. This may be achieved by irradiating the localized tumor site with radiation such as X-rays, UV-light, γ-rays or even microwaves. Alternatively, the tumor cells may be contacted with the agent by administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound such as, adriamycin, 5-fluorouracil, etoposide, camptothecin, actinomycin-D, mitomycin C, or more preferably, cisplatin. The agent may be prepared and used as a combined therapeutic composition, or kit, by combining it with the immunotherapeutic composition, as described above.

[0097] Agents that directly cross-link nucleic acids, specifically DNA, are envisaged to facilitate DNA damage leading to a synergistic, antineoplastic combination with the immunotherapeutic composition. Agents such as cisplatin, and other DNA alkylating agents may be used. Cisplatin has been widely used to treat cancer, with efficacious doses used in clinical applications of 20 mg/m² for 5 days every three weeks for a total of three courses. Cisplatin is not absorbed orally and must therefore be delivered via injection intravenously, subcutaneously, intratumorally, or intraperitoneally.

[0098] Agents that damage DNA also include compounds that interfere with DNA replication, mitosis, and chromosomal segregation. Such chemotherapeutic compounds include adriamycin, also known as doxorubicin, etoposide, verapamil, podophyllotoxin, and the like. Widely used in a clinical setting for the treatment of neoplasms, these compounds are administered through bolus injections intravenously at doses ranging from 25-75 mg/m² at 21 day intervals for adriamycin, to 35-50 mg/m for etoposide intravenously or double the intravenous dose orally.

[0099] Agents that disrupt the synthesis and fidelity of nucleic acid precursors and subunits also lead to DNA damage. As such a number of nucleic acid precursors have been developed. Particularly useful are agents that have undergone extensive testing and are readily available. As such, agents such as 5-fluorouracil (5-FU), are preferentially used by neoplastic tissue, making this agent particularly useful for targeting to neoplastic cells. Although quite toxic, 5-FU, is applicable in a wide range of carriers, including topical, however intravenous administration with doses ranging from 3 to 15 mg/kg/day being commonly used.

[00100] Other factors that cause DNA damage and have been used extensively include what are commonly known as γ-rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells. Other forms of DNA damaging factors are also contemplated such as microwaves and UV-irradiation. It is most likely that all of these factors effect a broad range of damage DNA, on the precursors of DNA, the replication and repair of DNA, and the assembly and maintenance of chromosomes. Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 weeks), to single doses of 2000 to 6000 roentgens. Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.

[00101] The skilled artisan is directed to "Remington's Pharmaceutical Sciences" 15th Edition, chapter 33, in particular pages 624-652. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.

[00102] The inventors propose that the local or regional delivery of the immuno therapeutic composition to patients with cancer will be a very efficient method for treating the clinical disease. Similarly, the chemo- or radiotherapy may be directed to a particular, affected region of the subject's body. Alternatively, systemic delivery of the immunotherapeutic composition and/or the agent may be appropriate in certain circumstances, for example, where extensive metastasis has occurred.

[00103] In addition to combining immunotherapies with chemo- and radiotherapies, it also is contemplated that combination with gene therapies will be advantageous. For example, any tumor-related gene conceivably can be targeted in combination with the immunotherapy, for example, p21, Rb, APC, DCC, NF-I, NF-2, BCRA2, pi 6, FHIT, WT-I, MEN-I, MEN-II, BRCAl, VHL, FCC, MCC, ras, myc, neu, raf, erb, src, fins, jun, trk, ret, gsp, hst, bcl and abl. [00104] Where clinical applications are contemplated, it will be necessary to prepare pharmaceutical compositions in a form appropriate for the intended application. Generally, this will entail preparing compositions that are essentially free of pyrogens, as well as other impurities that could be harmful to humans or animals. The phrase "pharmaceutically or pharmacologically acceptable" refers to entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human. Except insofar as any conventional media or agent is incompatible with the immunotherapeutic composition of the present invention, its use in therapeutic compositions is contemplated. Supplementary active ingredients also can be incorporated into the compositions.

[00105] The pharmaceutical forms suitable for use include sterile compositions. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The prevention of the action of microorganisms can be brought about by various antibacterial an antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

VI. KITS

[00106] Generally, kits comprise separate vials or containers for the various reagents or compositions, such as sutures, polymers, tumor lysates, immunostimulatory agents, antibodies, etc. The reagents are also generally prepared in a form suitable for preservation. The various vials or containers are often held in blow-molded or injection-molded plastics.

VII. EXAMPLES

[00107] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. EXAMPLE I

IMMUNOSTIMULATORY SUTURES

[00108] Studies indicate that using a bioactive PLGA suture loaded with tumor lysate derived from a neuroblastoma solid tumor that containing tumor specific and tumor associated antigens and CpG ODN (suture I), neuroblastoma does not recur or is inhibited. The inventors have undertaken SEM characterization of the sutures and release profiles of CpG and antigen. The tumor growth studies indicate that CpG in the sutures/matrices alone show positive result.

[00109] Immunostimulatory sutures (CpG loaded suture or suture II) were prepared by extruding a melted ( __70°C) mixture of PLGA (75:25 0.47 dL/g) pellets and lyophilized CpG ODN 1826 (FIG. 7A and FIG. 7B). Each piece of suture extrudate used in tumor suppression experiments was approximately 0.6 mm x 30 mm in dimensions with an average weight of 150 mg and loaded with 200 μg CpG ODN. Scanning electron microscopy images provided confirmation of the suture dimensions and showed that sutures had a solid morphology with no noticeable fractures or cracks (FIG. 7C). To measure the release profiles of CpG ODN, 450 mgs of CpG ODN loaded sutures were placed into 10 mL of phosphate buffered saline and placed on a plate shaker at 37°C. The supernatant was measured for release of CpG ODN at 260 nm on a UV spectrophotometer (Spectramax M5 Microplate reader, Molecular Device) every day and fresh medium replaced. CpG ODN was found to be released over 35 days at a sustained rate (FIG. 7d).

[00110] The inventors anticipate similar sustained release profiles in vivo as the primary mechanism of PLGA degradation is hydrolytic (Wang et al, 2004; Uhrich et al, 1999). By controlling the molecular weight and the chemical composition of the PLGA or other biodegradable polymers used to prepare the suture, the inventors can control the release of CpG ODN from weeks to over a year (Uhrich et al, 1999; Intra et al, 2008; Zhang et al, 2007). The inventors can also control the dimensions of the suture by adjustment of the draw rate which provides a further mechanism for controlling the release profiles and mechanical properties of the suture. Increasing the draw rate decreases the mean diameter of the suture extrudate. The draw rate, composition, and molecular weight of PLGA used in the tumor suppression experiments in this study were carefully selected to closely match the duration of tumor growth measurements. [00111] The murine subcutaneous neuroblastoma model is an ideal model for testing the potential use of the immunostimulatory suture as it reliably and aggressively recurs following resection (Ohashi et al., 2006). To evaluate the use of immunostimulatory sutures in preventing recurrence of tumor in the minimal residual disease setting, we tested 4 groups of mice in which the primary tumor was resected. These groups included: 1) wounds closed with polyglycolic acid Vicryl suture (Ethicon), 2) wounds closed with Vicryl at which time the site of resection was locally injected with 200 μg CpG ODN, 3) wounds closed with Vicryl and CpG ODN loaded suture implanted into the same mice remotely on the opposite side to the resection site and 4) wounds closed with Vicryl and CpG ODN loaded suture.

[00112] Syngeneic female A/J mice (6-8 weeks old; n=10 per group; Harlan Laboratories, Indianapolis, Ind) were anesthetized using halothane and inoculated subcutaneously with 1x10⁶ Neuro-2a (N2a, ATCC, Manassas, VA) murine neuroblastoma wild-type cells on day 1. Tumor growth was measured with calipers and once the tumors had grown between 5 to 10 mm in any dimension, the mice were anesthetized with intraperitoneal injections of ketamine (87 mg/kg) and xylazine (13 mg/kg) mixture and the tumors resected. FIG. 8 A shows that neuroblastoma recurrence and growth was suppressed most significantly in mice in which the excision site contained the immunostimulatory CpG ODN loaded suture. Complete prevention of any neuroblastoma recurrence was achieved in all mice in this group upto day 35 and 8 out of 10 mice had survived by the study end-point (FIG. 8B). In contrast, all other groups had a proportion of mice that had died by day 19. Mouse survival and tumor growth suppression using the CpG ODN loaded sutures was remarkably better than in all other groups (P<0.05), including mice in which the site of tumor excision was locally injected with 200 μg of CpG ODN in solution. This data highlights the enhanced protection generated against progressive minimal residual disease by providing sustained release of CpG ODN at the site of resection over prolonged periods of time. The two mice that died in the group treated with CpG ODN loaded sutures had relatively small tumors. It is therefore possible that death in these two mice was due to metastases in the internal organs and this requires further investigation. Previous studies have shown that frequent intra-tumoral injections of CpG ODN can lead to the rejection of neuroblastoma (Carpentier et al, 1999) but high concentrations of CpG ODN still have the potential to induce inflammation and systemic septic shock-like systems (Datta et al., 2003). This approach of using CpG ODN loaded sutures is very efficient at reducing residual disease that was otherwise deadly in the mice where wounds were closed with commercial Vicryl sutures. [00113] Of significant interest, mice that had the immunostimulatory CpG ODN loaded suture implanted remotely from the site of resection developed more aggressive tumor growth than control mice with tumor excision alone. This data clearly illustrates that the mechanism of CpG ODN action on rumor suppression is local and highlights the benefits of utilizing a suture to provide sustained local delivery of the CpG ODN. It is possible that remote delivery of the CpG ODN accelerates tumor growth because it delivers a danger signal that acts as a decoy for key immune cells otherwise needed at the site of resection but this theory requires further investigation.

[00114] Mice tolerated the CpG ODN loaded sutures and remained in good health as determined by the Body Condition Scoring Technique (Ullman-Cullere and Foltz, 1999). The mice appeared well conditioned. The vertebrae and dorsal pelvis were not prominent but palpable with slight pressure indicating a state of good health in the mice. In addition, no abnormal behavior, obvious neurological deficits or drowsiness was observed suggesting that CpG ODN loaded sutures that prevent local disease recurrence are not inducing paraneoplastic disease, which is consistent with the observations of Auf et al. (2002).

[00115] To identify the key subset of immune cells responsible for preventing neuroblastoma recurrence after tumor excision and CpG ODN suture therapy, we depleted mice (n=10 per group) of NK, CD4+, and CD8+ T-cells. The fastest tumor growth was observed in the group of mice depleted of NK cells (FIG. 8c; PO.001 in comparison to all other groups). These results suggest that prevention of tumor recurrence and suppression of tumor growth by immunostimulatory sutures at the site of excision is predominantly mediated by NK cells, which is consistent with the observations of others (Carpentier et al, 1999; Castriconi et al, 2007; Zeng et al, 2005; Castriconi et al, 2004). Depletion of CD4+ and CD8+ T-cells also increased tumor growth in comparison to the control group without cellular depletions. Mice depleted of CD8+ T-cells had marginally increased tumor growth in comparison to mice depleted of CD4+ T-cells but NK cell depletion had the greatest effect on enhancing tumor growth and reducing tumor survival (FIG. 8d). A number of studies have suggested that the CpG ODN induced NK activity is mediated through plasmacytoid dendritic cells (Liu et al, 2008) and that the induced NK activity is correlated with tumor growth suppression (Ballas, 2007; Ballas et al, 1996). A/J mice cured of neuroblastomas by treatment with CpG ODN have been reported to reject further tumor challenges suggesting that this approach can induce long-term protection but this requires further investigation for mice treated with CpG ODN loaded sutures (Carpentier et al, 1999). [00116] In summary, the inventors have developed an immunostimulatory suture that could have the dual function of closing the site of tumor excision and providing sustained localized delivery of an immunostimulatory ligand that prevents local tumor recurrence. Children with neuroblastoma frequently have a decent response to standard therapies where the tumor is reduced to a minimal residual disease, only to later fall victim to progressive residual disease (Ohashi et al, 2006). These pre-clinical results suggest that closing the tumor excision wound site using CpG ODN loaded sutures has the potential to result in significant improvements in patient survival rates. Sutures prepared from PLGA are FDA approved and have been used effectively in patients for many years. CpG ODN as an adjuvant is demonstrated to be safe in primate models and in phase 2/3 human clinical trials (Jones et al, 1999 and Coley Pharmaceutical Group/Pfizer). The suture preparation uses an extrusion process that is simple, reproducible and adaptable for loading of a wide variety of other antitumor molecules, cytokines, antigens, and immunostimulants in combination or alone. This immunostimulatory suture is therefore expected to have significant impact in a clinical setting.

Material and Methods

[00117] Neuroblastoma usually arises in the adrenal glands and is the most common extracranial solid malignancy of infancy and childhood. The mortality of neuroblastoma approaches 80-90% in children with the aggressive forms of the disease. High risk neuroblastoma has a dismal prognosis despite aggressive treatment with surgery and chemo- and radiation therapy. Chemotherapy cannot be increased further due to unacceptable toxicity. Patients often have a favorable initial response to standard therapies in which the tumor is reduced to minimal residual disease, but unfortunately, such children frequently succumb to progressive residual disease. A suture/matrix for use after the neuroblastoma is resected and the site is sutured has been developed.

[00118] Suture Preparation. Suture I: Biodegradable PLGA sutures were loaded with tumor lysate derived from a neuroblastoma solid tumor that containing tumor specific and tumor associated antigens, and CpG ODN were prepared. Specifically, sutures were prepared by loading a mixture of powdered PLGA polymer, dye, CpG ODN and lyophilized tumor lysate into a hopper of a polymer extruder. The hopper was initially loaded with PLGA powder alone, followed by the mixture with the dye. The mixture was slightly heated before being forced through an extruder. In this way, the sutures being used had both the CpG ODN and the tumor lysate within the polymer matrix. [00119] Suture II: To prepare the immunostimulatory suture, the inventors loaded ground up polylactic acid-co-glycolic acid (PLGA 75:25 0.47 dL/g, Absorbable Polymers International, Pelham, AL) pellets and CpG ODN 1826 (5'-TCCATGACGTTCCTGACGTT- 3' (SEQ ID NO:1), Coley Pharmaceutical Group, Wellesley, MA) that was endotoxin free (<0.03 Eu/mL; BioWhittaker, Walkersville, MD) into a Dynisco extruder hopper and sutures were extruded from a melted ( ≤70°C) mixture of PLGA pellets and lyophilized CpG ODN.

[00120] Sutures of the invention could be evaluated for their ability to release CpG and proteins into a physiological medium. Previous studies conducted by the inventors on entrapping ovalbumin and QG within a polymer matrix have shown that the PLGA matrices follow a burst release followed by a more sustained release over time. Other studies have also shown that when CpG ODN and tumor proteins are released from polymeric matrices, they stimulated strong IgGl and TgG2a production against the antigen, upregulated IFN-γ production, whilst IL-IO (an immunosuppressive molecule) and IL-5 did not increase.

[00121] Scanning Electron Microscopy. Samples for scanning electron microscopy (SEM, Hitachi S-4000) were prepared by coating suture samples on steel stubs with approximately 5 nm of gold by ion beam evaporation using a sputter coater (E550 Emitech sputter coater) set at 10 mA for 10 seconds prior to examination in the SEM operated at 5 kV accelerating voltage.

[00122] Murine Tumor Cell Lines and model of minimal residual disease. The tumors can be tested in a pre-established gross tumor model and a model of minimal residual disease. The neuroblastoma model was chosen because it is particularly useful for studying the concept because all mice that undergo resection will die of recurrent disease using this model, which makes this an ideal model for studying the effect of adjuvant therapies on microscopic residual disease.

[00123] The Neuro-2a (NZa), a murine Neuroblastoma wild-type cell line, was purchased from American Type Culture Collection. The cells were cultured in vitro in Minimal Essential Medium (GIBCO, Grand Island, NY), supplemented with 10% Fetal Bovine Serum, 1% Penicillin-streptomycin (10,000 U/ml), 10 mmol Sodium Pyruvate, 100 mmol Nonessential Amino Acid (GIBCO) and 0.75g sodium Bicarbonate (Fisher Scientific) and were free of Mycoplasma. [00124] Neuro-2a (I x 10⁶) wild type cells were inoculated subcutaneously (SQ) on day 1. A visible tumor was usually observed about 6 days after tumor challenge. Caliper measurements of tumor development and growth were monitored and documented at least 3 to 4 times a week, and volumes were calculated as (width)² x length x 0.5. At a specified time-point, when all the tumors had reached a size of about 5 to 10 mm in any dimension the tumors were resected. The primary tumor was excised in each case without lymph node dissection. Mice were randomized before assigning to groups 1-4. The average maximum size of the tumors in any dimension for each group prior to resection were 1) 6.3 +/- 3.2 mm, 2) 8.1 +/- 1.6 mm, 3) 5.9 +/- 3.9 mm, and 4) 7.4 +/- 2.3 mm. The suture I (PLGA suture loaded with tumor lysate derived from a neuroblastoma solid tumor and CpG ODN) was then used at the site of resection. Alternatively, the site of excision or resection was then closed using either the immunostimulatory CpG ODN loaded sutures (suture II) and/or Vicryl and tumor growth development monitored. Tumor regrowth following resection and use of immunostimulatory sutures is typically assessed using caliper measurements and measurement of immune responses generated from the sutures. All experiments were repeated at least once.

[00125] Anesthetic agents and animal care: Female A/J mice (6-8 weeks old; Harlan Laboratories, Indianapolis, IN) were anesthetized using halothane inhalation (Halocarbon, River Edge, NJ) during inoculation. Mice undergoing tumor resections were anesthetized with intra-peritoneal Ketamine 987 mg/kg) and Xylazine (13 mg/kg) mixture and tumors were resected under clean conditions. All of the animals were housed under standard conditions in accordance with University of Iowa's animal care and use committee, which follows the USPHS guide for the care and use of animals. Mice were sacrificed if tumor size was greater than 2.5 cm in any dimension of if mice assumed a "sick mouse posture."

[00126] Depletion of Effector Cells. NK cells, CD4+, and CD8+ T-cells were depleted by injecting mice intraperitoneally with 100 μg of 2.43 or GK1.5/mouse for 3 consecutive days before the tumor was resected and thereafter every other day for the duration for the experiment. The anti-NK antibody anti-asialo GMl (asGMl; Wako Bioproducts, Richmond, VA) was injected once at 25 μg/mouse 3 days before tumor resection and thereafter every 4 days at 25 μg/mouse. The control antibody used for the group with no depletions was a nonspecific rat IgG2b (SFR8-B6, ATCC, Manassas, VA). All injection volumes were 100 μl. Verification of depletion of T-cells was determined using flow cytometric analysis of spleen cells stained with 2.43 or GKl.5 followed by FITC-labeled goat antibody to rat IgG. Greater than 99% depletion of the desired populations was observed. All experiments were repeated at least once.

[00127] Statistical Analysis. The statistical analyses focused on different wound closure strategies following tumor resection and the effects on tumor recurrence and progression. The primary outcomes of interest were time to death and tumor growth over time. The log-rank test was used to compare the survival times between vaccination groups, and Kaplan-Meier survival functions. Tumor size (cm³) was regularly measured throughout the experiments, resulting in repeated measurements across time for each mouse. Mixed linear regression models were used to estimate and compare the group-specific tumor growth curves. Continuous first-order autoregressive structures were specified in the models to account for the within-subject correlation. In both the survival and growth curve analyses, statistically significant global tests of equality across groups were followed up with pairwise comparisons to identify specific group differences.

* * * * * * * * *

[00128] All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents that are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

REFERENCES

The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference:

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Claims

1. A surgical suture comprising (a) a biocompatible filament and (b) an immunostimulatory agent, wherein said immunostimulatory agent is present throughout the width of one or more sections of the biocompatible filament.

2. The suture of claims 1, wherein said immunostimulatory agent comprises bacterial cell wall, LPS₅ bacterial DNA, viral RNA, CpG oligonucleotides, double-stranded RNA, β-glucan, zymosan™, GM-CSF, IL-2, IL-6, IL-7, IL-15, IFN-γ, or IFN-α.

3. The suture of claim 2, wherein the immunostimulatory agent is CpG oligonucleotide.

4. The suture of claim 1 , further comprising one or more tumor cell antigens.

5. The suture of claims 4, wherein the one or more tumor cell antigen is a tumor cell lysate.

6. The suture of claim 5, wherein the tumor cell lysate is derived from a breast cancer cell, a head and neck cancer cell, a lung cancer cell, a stomach cancer cell, an esophageal cancer cell, a skin cancer cell, a colon cancer cell, an ovarian cancer cell, a prostate cancer cell, a testicular cancer cell, a uterine cancer cell, a cervical cancer cell, a pancreatic cancer cell, or a liver cancer cell.

7. The suture of claims 1, wherein said biocompatible filament is biodegradable .

8. The suture of claim 7, wherein said biodegradable filament is a polymer.

9. The suture of claim 8, wherein the polymer is selected from the group consisting of polyethylene glycol, poly(lactic acid), poly(glycolic acid), copolymers of lactic and glycolic acid, copolymers of lactic and glycolic acid with polyethylene glycol, poly(ε- caprolactone), poly(3-hydroxybutyrate), poly(p-dioxanone), polypropylene fumarate, poly(orthoesters), polyol/diketene acetals addition polymers, poly(sebacic anhydride) (PSA), poly(carboxybiscarboxyphenoxyphenoxy hexone (PCPP) poly[bis (p- carboxypheonoxy) methane] (PCPM), copolymers of SA, CPP and CPM, poly(amino acids), poly(pseudo amino acids), polyphosphazenes, derivatives of poly[(dichloro)phosphazenes] and poly[(organo) phosphazenes], poly-hydroxybutyric acid, and S-caproic acid.

10. The composition of claim 9, wherein the polymer is polylactide-co-glycolide and the immunostimulatory agent is CpG oligonucleotide.

11. The composition of claim 9, wherein the polymer is polylactic acid and polyethylene glycol, and the immunostimulatory agent is CpG.

12. The composition of claim 10, further comprising GM-CSF.

13. The composition of claim 11 , further comprising GM-CSF.

14. The composition of claim 1 , further comprising a non-tumor protein.

15. The composition of claim 14, wherein the non-tumor protein is ovalalbumin.

16. A method of treating cancer in a subject comprising the steps of:

(a) surgically resecting a tumor; and

(b) contacting the resection site with a biologically active suture comprising an immunostimulatory agent.

17. The method of claim 16, wherein said suture further comprises a plurality of tumor cell antigens.

18. The method of claim 16, wherein said suture is biodegradable.

19. The method of claim 16, wherein said suture is a silk, an elastin, a chitin, a chitosan, a poly(d-hydroxy acid), a poly(anhydrides), or a poly(athoesters) suture.

20. The method of claim 16, wherein said suture comprises polyethylene glycol, polyQactic acid), poly(glycolic acid), copolymers of lactic and glycolic acid, copolymers of lactic and glycolic acid with polyethylene glycol, poly(ε-caprolactone), poly(3-hydroxybutyrate), poly(p-dioxanone), polypropylene fumarate, poly(ortho esters), polyol/diketene acetals addition polymers, poly(sebacic anhydride) (PSA), poly(carboxybiscarboxyphenoxyphenoxy hexone (PCPP) poly[bis (p- carboxypheonoxy) methane] (PCPM), copolymers of SA, CPP and CPM, poly(amino acids), poly(pseudo amino acids), polyphosphazenes, derivatives of poly[(dichloro)phosphazenes] and poly[(organo) phosphazenes], poly-hydroxybutyric acid, or S-caproic acid.

21. The method of claim 16, wherein said immunostimulatory agent comprises bacterial cell wall, LPS, bacterial DNA, viral RNA, CpG oligonucleotides, double-stranded RNA, β-glucan, zymosan™, GM-CSF, IL-2, IL-6, IL-7, IL-15, IFN-γ, or IFN-α.

22. The method of claim 21, wherein the immunostimulatory agent is a CpG oligonucleotide.

23. The method of claim 17, wherein said plurality of tumor cell antigens are in the form of a tumor cell lysate or a fraction of tumor cell lysate.

24. The method of claim 23, wherein the tumor cell lysate is derived from a breast cancer cell, a head and neck cancer cell, a lung cancer cell, a stomach cancer cell, an esophageal cancer cell, a skin cancer cell, a colon cancer cell, an ovarian cancer cell, a prostate cancer cell, a testicular cancer cell, a uterine cancer cell, a cervical cancer cell, a pancreatic cancer cell, or a liver cancer cell.

25. The method of claim 23, wherein the tumor cell lysate is derived from a neuroblastoma, a WiIm' s tumor, a rhabdoid tumor, a sarcoma (osteogenic or non- osteogenic), a hepatoblastoma, a rhabdomyosarcoma, a lymphoma, or a leukemia.

26. The method of claim 16, wherein said cancer is a neuroblastoma, a melanoma, a WiIm' s tumor, a rhabdoid tumor, a sarcoma (osteogenic or non-osteogenic), a hepatoblastoma, a rhabdomyosarcoma, a lymphoma, a leukemia, a breast cancer, a head and neck cancer, a lung cancer, a stomach cancer, an esophageal cancer, a skin cancer a colon cancer, an ovarian cancer, a prostate cancer, a testicular cancer, a uterine cancer, a cervical cancer, a pancreatic cancer, or a liver cancer.

27. A suture kit comprising composition comprising (a) a biocompatible filament and (b) a plurality of tumor cell antigens and an immunostimulatory agent encapsulated in said biocompatible filament, the composition being disposed in a discrete container.