Abstract: The present invention discloses that an intratumoral injection of: i) glycolipids with ?-gal epitope; ii) gene vectors comprising an ?1,3galactosyltransferase gene; or iii) a mixture of ?1,3galactosyltransferase, neuraminidase, and uridine diphosphate galactose results in tumor regression and/or destruction. Binding of the natural anti-Gal antibody to de novo expressed tumoral ?-gal epitopes induces inflammation resulting in an anti-Gal antibody mediated opsonization of tumor cells and their uptake by antigen presenting cells. These antigen presenting cells migrate to draining lymph nodes and activate tumor specific T cells thereby converting the treated tumor lesions into in situ autologous tumor vaccines. This therapy can be applied to patients with multiple lesions and in neo-adjuvant therapy to patients before tumor resection.

Abstract: The present invention is related to the field of wound healing or tissue regeneration due to disease (i.e., for example, cardiovascular diseases, osetoarthritic diseases, or diabetes). In particular, the present invention provides compositions and methods comprising molecules with linked ?-gal epitopes for induction of an inflammatory response localized within or surrounding damaged tissue. In some embodiments, the present invention provides treatments for tissue repair in normal subjects and in subjects having impaired healing capabilities, such as diabetic and aged subjects.

Abstract: The present invention relates to the microbial immunogens engineered to bear ?-gal epitope(s) for induction of potent humoral and cellular immune responses when administered to subjects having anti-Gal antibodies. In one embodiment, the present invention provides compositions and methods for propagating influenza virus in human, ape, Old World monkey or bird cells that have been engineered to express an ?1,3galactosyltransferase (? 1,3GT) gene to produce virions bearing hemagglutinin molecules containing ?-gal epitopes, to increase the immunogenicity of the influenza virus. In another embodiment, the present invention provides fusion proteins between influenza virus hemagglutinin and a microbial peptide or protein of interest, and enzymatic processing of this fusion protein to carry ?-gal epitopes, to increase the immunogenicity of the microbial peptide or protein of interest.

Abstract: The present invention is related to the field of wound healing or tissue regeneration due to disease (i.e., for example, cardiovascular diseases, osteoarthritic diseases, or diabetes). In particular, the present invention provides compositions and methods comprising molecules with linked ?-gal epitopes for induction of recruitment of macrophages localized within or surrounding damaged tissue. The recruited macrophages and stem cells promote the repair and regeneration of the treated injured tissue. In some embodiments, the present invention provides treatments for tissue repair in normal subjects and in subjects having impaired healing capabilities, such as diabetic and aged subjects. In some embodiments, the present invention provides treatments for injured tissues such as brain, peripheral nerve, muscle, cartilage, bone, gastrointestinal tract and dysfunctional endocrine tissues.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic bone xenograft (X) for implantation into a defect (D) located in a bone portion (10) of a human. The invention further provides methods for preparing a bone xenograft (X) by removing at least a portion of bone from a non-human animal to provide a xenograft (X); washing the xenograft (X) in saline and alcohol; and subjecting the xenograft (X) to at least one of the treatments including exposure to ultraviolet radiation, immersion in alcohol, ozonic, and freeze/thaw cycling. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment, and digestion of the carbohydrate moieties of the xenograft (X) with a glycosidase followed by treatment for sialylation.

Abstract: The present invention discloses that an intratumoral injection of: i) glycolipids with ?-gal epitope; ii) gene vectors comprising an ?1,3galactosyltransferase gene; or iii) a mixture of ?1,3galactosyltransferase, neuraminidase, and uridine diphosphate galactose results in tumor regression and/or destruction. Binding of the natural anti-Gal antibody to de novo expressed tumoral ?-gal epitopes induces inflammation resulting in an anti-Gal antibody mediated opsonization of tumor cells and their uptake by antigen presenting cells. These antigen presenting cells migrate to draining lymph nodes and activate tumor specific T cells thereby converting the treated tumor lesions into in situ autologous tumor vaccines. This therapy can be applied to patients with multiple lesions and in neo-adjuvant therapy to patients before tumor resection.

Abstract: A novel noninvasive immunological method for early detection of immune reactivity, primarily antibody reactivity, in heart transplant recipients prior to tissue damage or rejection is disclosed. The method utilizes donor tissue obtained before transplant and processed to obtain an antigen preparation comprising homogenized, fragmented donor cardiac tissue, including the posterior wall of the right atrium and the joining portion of the vena cava. This donor tissue is normally discarded before transplant. The methods include ELISA or light emitting immunoassays for measuring antibody binding. These methods are also applicable to allografts of other organs such as kidney. Such early detection of these immune responses enables a treating physician to modify the immunosuppressive treatment in order to prevent episodes of immune rejection and thus, to prevent damage of the transplanted organ.

Abstract: A method for preparing a soft tissue xenograft includes the steps of removing at least a portion of a soft tissue from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol: subjecting the xenograft to cellular disruption treatment; and either digesting the xenograft with a glycosidase or glycosidase digestion followed by treatment for sialylation. A soft tissue xenograft for implantation into a human includes a portion of a soft tissue from a non-human animal, wherein the portion has extracellular matrix and substantially only dead cells. The matrix and dead cells have substantially no surface &agr;-galactosyl moieties and have sialic acid molecules linked to at least a portion of surface carbohydrate moieties. Each of the xenografts of the invention has substantially the same mechanical properties as a corresponding native soft tissue.

Abstract: The present invention provides compositions and methods for inducing immune tolerance to one or more specific antigens in a host mammal. Generally, the methods involves engineering white blood cells, in vitro, to express an antigen which is not native to the host mammal. Cells engineered ex vivo are then introduced into the host mammal to induce immune tolerance to the expressed antigen.

Abstract: A method for preparing a soft tissue xenograft includes the steps of removing at least a portion of a soft tissue from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; subjecting the xenograft to cellular disruption treatment; and either digesting the xenograft with a glycosidase or glycosidase digestion followed by treatment for sialylation. A soft tissue xenograft for implantation into a human includes a portion of a soft tissue from a non-human animal, wherein the portion has extracellular matrix and substantially only dead cells. The matrix and dead cells have substantially no surface &agr;-galactosyl moieties and have sialic acid molecules linked to at least a portion of surface carbohydrate moieties. Each of the xenografts of the invention has substantially the same mechanical properties as a corresponding native soft tissue.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic bone xenograft (X) for implantation into a defect (D) located in a bone portion (10) of a human. The invention further provides methods for preparing a bone xenograft (X) by removing at least a portion of bone from a non-human animal to provide a xenograft (X); washing the xenograft (X) in saline and alcohol; and subjecting the xenograft (X) to at least one of the treatments including exposure to ultraviolet radiation, immersion in alcohol, ozonic, and freeze/thaw cycling. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment, and digestion of the carbohydrate moieties of the xenograft (X) with a glycosidase followed by treatment for sialylation.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic ligament or tendon xenograft for implantation into humans. The invention further provides a method for preparing a ligament xenograft by removing at least a portion of a ligament from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; subjecting the xenograft to at least one treatment selected from the group consisting of exposure to ultraviolet radiation, immersion in alcohol, ozonation, freeze/thaw cycling, and optionally chemical crosslinking. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment and either digestion of the carbohydrate moieties of the xenograft with a glycosidase in a range of about 1 mU/ml to about 1000 U/ml or glycosidase digestion followed by treatment for sialylation. The invention also provides articles of manufacture produced by one or more of the above-identified methods of the invention.

Abstract: Toxin complexes and methods for preventing immune rejection of xenografts are provided. Cell toxins are coupled to carriers that display ligands which allow the complex to be recognized by antigen binding sites (i.e., specific B cell and T cell receptors) on the lymphocytes as the target cells responsible for the specific immune response. The toxin complexes interact with the specific receptors (antigen binding sites) on lymphocytes, and are internalized by these target cells. Once internalized, the toxin complex dissociates to release the cell toxin, which destroys the target cell. In one embodiment, the target cells are B lymphocytes and T lymphocytes, the antigen binding sites are anti-Gal B cell receptors and T cell receptors which interact with &agr;-gal epitopes displayed on an &agr;1-acid glycoprotein carrier. In other embodiments, the toxin complex is directed by the corresponding ligands to target cells responsible for autoimmune diseases in which the specificity of the autoantibodies is defined.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic ligament or tendon xenograft for implantation into humans. The invention further provides a method for preparing a ligament xenograft by removing at least a portion of a ligament from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; subjecting the xenograft to at least one treatment selected from the group consisting of exposure to ultraviolet radiation, immersion in alcohol, ozonation, freeze/thaw cycling, and optionally chemical crosslinking. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment and either digestion of the carbohydrate moieties of the xenograft with a glycosidase in a range of about 1 mU/ml to about 1000 U/ml or glycosidase digestion followed by treatment for sialylation. The invention also provides articles of manufacture produced by one or more of the above-identified methods of the invention.

Abstract: The invention encompasses methods and compositions for inducing an immune response in an anti-Gal synthesizing animal including viral and tumor antigens manipulated to express &agr;-galactosyl epitopes.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic ligament or tendon xenograft for implantation into humans. The invention further provides a method for preparing a ligament xenograft by removing at least a portion of a ligament from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; subjecting the xenograft to at least one treatment selected from the group consisting of exposure to ultraviolet radiation, immersion in alcohol, ozonation, freeze/thaw cycling, and optionally chemical crosslinking. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment and either digestion of the carbohydrate moieties of the xenograft with a glycosidase in a range of about 1 mU/ml to about 1000 U/ml or glycosidase digestion followed by treatment for sialylation. The invention also provides articles of manufacture produced by one or more of the above-identified methods of the invention.

Abstract: The invention provides a substantially non-immunogenic ligament or tendon xenograft for implantation into humans. The invention further provides a method for preparing a ligament xenograft by removing at least a portion of a ligament from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; subjecting the xenograft cellular disruption treatment, such as exposure to ultraviolet radiation, immersion in alcohol, ozonation, freeze/thaw cycling, and optionally chemical crosslinking. In addition to or in lieu of the above treatments, the methods include digestion of the carbohydrate moieties of the xenograft with a glycosidase, preferably in a range of about 1 mU/ml to about 1000 U/ml, followed by treatment for sialylation. The invention also provides articles of manufacture produced by one or more of the above-identified methods of the invention.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic articular cartilage xenograft for implantation into humans. The invention further provides a method for preparing an articular cartilage xenograft by removing at least a portion of an articular cartilage from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; subjecting the xenograft to at least one treatment selected from the group consisting of exposure to ultraviolet radiation, immersion in alcohol, ozonation, freeze/thaw cycling, and optionally to chemical crosslinking. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment and either digestion of the carbohydrate moieties of the xenograft with a glycosidase in a range of about 1 mU/ml to about 1000 U/ml or glycosidase digestion followed by treatment for sialylation.

Abstract: The invention provides an article of manufacture comprising a substantially non-immunogenic knee meniscal xenograft for implantation into humans. The invention further provides methods for preparing a knee meniscal xenograft by removing at least a portion of a meniscus from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol; and subjecting the xenograft to at least one treatment selected from the group consisting of exposure to ultraviolet radiation, immersion in alcohol, ozonation, and freeze/thaw cycling. In addition to or in lieu of the above treatments, the methods include a cellular disruption treatment and either digestion of the carbohydrate moieties of the xenograft with a glycosidase in a range of about 1 mU/ml to about 1000 U/ml or glycosidase digestion followed by treatment for sialylation. The invention also provides articles of manufacture produced by one or more of the above-identified methods of the invention.

Abstract: The invention encompasses methods and compositions for inducing an immune response in an anti-Gal synthesizing animal including viral and tumor antigens manipulated to express .alpha.-galactosyl epitopes.