Infectious Disease Vaccines

Stress Protein Compositions and Methods for Prevention and Treatment of Cancer and Infectious Disease

U.S. Patent Number: 6,984,384; 7,378,096 (CIP)
Date Issued: January 10, 2006; May 27, 2008 (CIP)

Summary: Pharmaceutical compositions comprising a stress protein complex and related molecules encoding or cells presenting such a complex are provided. The stress protein complex comprises an hsp110 or grp170 polypeptide complexed with an immunogenic polypeptide. The immunogenic polypeptide of the stress protein complex can be associated with a cancer or an infectious disease. The pharmaceutical compositions of the invention can be administered to a subject, thereby providing methods for inhibiting M. tuberculosis-infection, for inhibiting tumor growth, for inhibiting the development of a cancer, and for the treatment or prevention of infectious disease. The invention further provides a method for producing T cells directed against a tumor cell or a M. tuberculosis-infected cell, wherein a T cell is contacted with an APC that is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with a tumor or with the M. tuberculosis-infected cell. Included in the invention are T cells produced by this method and a pharmaceutical composition comprising such T cells. The T cells can be contacted with a M. tuberculosis-infected cell in a method for killing a M. tuberculosis-infected cell, or with a tumor cell in a method for killing a tumor cell.

Detail: The invention provides a pharmaceutical composition comprising a stress protein complex. The stress protein complex comprises an hsp110 or grp170 polypeptide and an immunogenic polypeptide. In some embodiments, the hsp110 or grp170 polypeptide is complexed wit h the immunogenic polypeptide, for example, by non-covalent interaction or by covalent interaction, including a fusion protein. In some embodiments, the complex is derived from a tumor. In other embodiments, the complex is derived from cells infected with an infectious agent. The immunogenic polypeptide of the stress protein complex can be associated with a cancer or an infectious disease. The stress protein complex of the invention can further include additional stress polypeptides, including members of the hsp70, hsp90, grp78 and grp94 stress protein families. In one embodiment, the stress protein complex comprises hsp110 complexed with hsp70 and/or hsp25. The invention additionally provides a pharmaceutical composition comprising a first polynucleotide encoding an hsp 110 or a grp170 polypeptide and a second polynucleotide encoding an immunogenic polypeptide. In some embodiments involving first and second polynucleotides, the first polynucleotide is linked to the second polynucleotide. The pharmaceutical compositions of the invention can further comprise a physiologically acceptable carrier and/or an adjuvant. The efficacy of a pharmaceutical composition can further comprise GM-CSF-secreting cells. Alternatively, GM-CSFsecreting cells can be co-administered with a pharmaceutical composition of the invention, by administration before, during or after administration of the pharmaceutical composition. The use of GM-CSF-secreting cells enhances the efficacy of the pharmaceutical composition. In some embodiments, the complex is purified from a tumor or from cells infected with an infectious agent. In such embodiments, the stress polypeptide, as purified, is complexed with one or more immunogenic polypeptides. The binding of the stress polypeptide to the immunogenic polypeptide can be altered and/or enhanced by stress, such as by exposure to heat, anoxic and/or ischemic conditions, or proteotoxic stress. In particular, a stress protein complex of the invention can comprise a stress polypeptide complexed with an immunogenic polypeptide, wherein the complex has been heated. Such heating, particularly wherein the stress polypeptide comprises a heat-inducible stress protein, can increase the efficacy of the stress protein complex as a vaccine. Examples of heat-inducible stress proteins include, but are not limited to, hsp70 and hsp 110. In some embodiments, the immunogenic polypeptide is known. The immunogenic polypeptide is a known molecule, the immunogenic polypeptide can be provided in admixture with the stress polypeptide, or as a complex with the stress polypeptide. The hsp 110 or grp170 polypeptide can be complexed with the immunogenic polypeptide by non-covalent binding. Alternatively, the complex can comprise a fusion protein, wherein the stress polypeptide is linked to the immunogenic polypeptide. Examples of immunogenic polypeptides include, but are not limited to, antigens associated with cancer or infectious disease, such as the breast cancer antigen her2/neu or the Mycobacterium tuberculosis antigens Mtb8.4 and Mtb39. Where the immunogenic polypeptide is unknown, it can be obtained incidentally to the purification of the stress polypeptide from tissue of a subject having cancer or an infectious disease. Also provided is a pharmaceutical composition comprising an antigen-presenting cell (APC) modified to present an hsp 110 or grp170 polypeptide and an immunogenic polypeptide. Alternatively, the APC can be modified to present an immunogenic polypeptide obtained by purification of hsp 110 or grp170 from disease cells, including cancer cells and cells infected with an infectious agent. Preferably, the APC is a dendritic cell or a macrophage. The APC can be modified by various means including, but not limited to, peptide loading and transfection with a polynucleotide encoding an immunogenic polypeptide. The pharmaceutical compositions of the invention can be administered to a subject, thereby providing methods for inhibiting M. tuberculosis-infection, for inhibiting tumor growth, for inhibiting the development of a cancer, and for the treatment or prevention of cancer or infectious disease. The invention further provides a method for producing T cells directed against a tumor cell. The method comprises contacting a T cell with an antigen presenting cell (APC), wherein the APC is modified to present an hsp 110 or grp170 polypeptide and an immunogenic polypeptide associated with the tumor cell. Such T cells can be used in a method for killing a tumor cell, wherein the tumor cell is contacted with the T cell. Likewise, the invention provides a method for producing T cells directed against a M. tuberculosis-infected cell, wherein a T cell is contacted with an APC that is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with the M. tuberculosis-infected cell. Included in the invention are T cells produced by this method and a pharmaceutical composition comprising such T cells. The T cells can be contacted with a M. tuberculosis-infected cell in a method for killing a M. tuberculosis-infected cell The T cells can be CD4+ or CD8+. The invention also provides a method for removing tumor cells from a biological sample. The method comprises contacting a biological sample with a T cell of the invention. In a preferred embodiment, the biological sample is blood or a fraction thereof. Also provided is a method for inhibiting tumor growth in a subject. The method comprises incubating CD4+ and/or CD8+ T cells isolated from the subject with an antigen presenting cell (APC), wherein the APC is modified to present an hsp 110 or grp 170 polypeptide and an immunogenic polypeptide associated with the tumor cell such that T cells proliferate. The method further comprises administering to the subject an effective amount of the proliferated T cells, and thereby inhibiting tumor growth in the subject. In an alternative embodiment, the method for inhibiting tumor growth in a subject comprises incubating CD4+ and/or CD8+ T cells isolated from the subject with an antigen presenting cell (APC), wherein the APC is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with the tumor cell such that T cells proliferate, cloning at least one proliferated cell, and administering to the patient an effective amount of the cloned T cells, thereby inhibiting tumor growth in the subject.

Compositions and Methods for Prevention and Treatment of Fungal Diseases

U.S. Patent: Pending
Application Number: 11/554,022

Summary: The invention relates to various pharmaceutical compositions that can be used as active or passive vaccines for the treatment or prevention of fungal disease. Methods for prevention and treatment of infectious and allergic fungal diseases in subjects using the pharmaceutical compositions of the present invention are also disclosed.

Detail: The invention relates to various pharmaceutical compositions that can be used as active or passive vaccines for the treatment or prevention of fungal disease. Methods for prevention and treatment of infectious and allergic fungal diseases in subjects using the pharmaceutical compositions of the present invention are also disclosed. The use of a natural, autologous stress protein or polypeptide adjuvant has significant clinical importance, because effective adjuvants and effective therapeutics for fungal diseases are currently lacking. The invention provides both active vaccines (containing either (i) a human stress protein or polypeptide complexed to a relevant immunogenic fungal polypeptide or (ii) a nucleic acid vaccine that encodes these polypeptides) and passive vaccines (containing activated antigen presenting cells). These vaccines will afford a highly potent, yet safe, antifungal vaccine suitable for prophylaxis and therapy in humans as well as in other animals.

Method of Raising an Immune Response with an Anti-idiotypic Antibody Having Correspondence with Human Hepatitis B Surface Antigen

U.S. Patent Number: 5,531,990; 5,668,253 (DIV); 5,744,135 (CON); 5,856,087 (CON)
Date Issued: July 2, 1996; September 16, 1997 (DIV); April 28, 1998 (CON); January 5, 1999 (CON)

Summary: The invention comprises an anti-idiotypic antibody designated 2F10 and permitted variants thereof, which have antigenic properties similar to the group specific “a” determinant of human hepatitis B surface antigen HBsAg and have at least partial but not complete homology with such surface antigen. The invention further comprises a peptide having a chain comprising the amino acid residues Ala Val Tyr Tyr Cys Thr Arg Gly Tyr His Gly Ser Ser Leu Tyr and permited variants thereof, which, like 2F10, have antigenic properties similar to the group specific “a” determinant of human hepatitis B surface antigen HBsAg and have at least partial, but not complete, homology with said surface antigen. The amino acid sequence is found in and forms a part of 2F10. The shorter peptide chain comprising the amino acid residues Gly Tyr His Gly Ser Ser Leu Tyr and permited variants thereof, also have antigenic properties similar to the group specific “a” determinant of human hepatitis B surface antigen HBsAg and have at least partial, but not complete, homology with said surface antigen.

Detail: A method for causing an immune response to human Hepatitis B surface antigen which consists essentially of raising an immune response to an antiidiiotypic antibody designated 2F10 produced by a monoclonal antibody having ATCC deposit number HB 11966 or to a 15 amino acid sequence of the heavy chain hypervariable region of the antiiodotypic antibody 2F10, said sequence being partially homologous with the group specific “a” determinant of Hepatitis B surface antigen, wherein said 15 amino acid sequence can duplicate the B and T cell stimulatory activity of said antiiodotypic antibody 2F10 and Hepatitis B surface antigen.

Method of Immunizing Against Hepatitis B Virus

U.S. Patent Number: 6,319,501
Date Issued: November 20, 2001

Summary: A method for immunizing a human against hepatitis B virus comprising administering to the human a vaccine comprising a hepatitis B virus surface antigen, wherein included in the vaccine is one or more antigens of non-permitted variant sequences within residues S(139-147) of the hepatitis B virus surface antigen.

Detail: The invention concerns an improved method for immunizing against hepatitis B virus comprising administering to a human a vaccine containing a hepatitis B virus surface antigen, e.g., the entire hepatitis B virus S protein or one or more portions of the entire S protein, wherein the improvement comprises including in such vaccine one or more antigens of nonpermitted (not tolerated) variant sequences within residues S(139-147) (CTKPSDGNC) of the hepatitis B virus surface antigen. The antigen may be the entire S protein sequence or one or more portions thereof. Such antigen will contain one or more replacements within residues S(139-147). The invention is also directed to a method for overcoming the detrimental immunological effects of mutations in the S(139-147) sequence of hepatitis B virus surface antigen comprising including in an immunogenic hepatitis B vaccine for administration to a human one or more antigens of non-permitted variant sequences within residues S(139-147) of the hepatitis B virus surface. The invention also relates to an improved hepatitis B virus vaccine, comprising adding to the vaccine (containing either the entire hepatitis B virus S protein or one or more portions thereof) one or more antigens of non-permitted variant sequences within residues S(139-147) of the hepatitis B virus surface antigen. More particularly, the above described inventive methods and vaccines involve the utilization of a non-permitted variant sequence with an amino acid replacement at one or more of positions 142, 143, 144, 145 or 146 of S(139-147). Still more particularly, the inventive methods and vaccines involve the employment of a non-permitted variant sequence with one or more of the following amino acid replacements: 142 P to S, 143 T(S) to M, 144 D to N or E, 145 G to A or R, or 146 N to D.

Expression of Immunogenic Hepatitis B Surface Antigens in Transgenic Plants

U.S. Patent Number: 6,551,820
Date Issued: April 22, 2003

Summary: Plant expression vectors comprising at least two expression cassettes are provided which function to reduce transcriptional silencing of polynucleotide expression. Further, novel plant expression vectors for expression of immunogenic polypeptides, including HBsAg, are provided. The plant expression vectors can be used to produce immunogenic polypeptides, including HBsAg, in edible plant tissues. The edible plant tissues can be used to elicit an immune response in humans and animals when the plant tissues are consumed.

Detail: It is an object of the invention to provide plant expression vectors comprising at least two expression cassettes that function to reduce transcriptional silencing of polynucleotide expression. It is another object of the invention to provide novel plant expression vectors for expression of immunogenic polypeptides, including HBsAg. The plant expression vectors can be used to produce immunogenic polypeptides, including HBsAg, in edible plant tissues. It is a further object of the invention to provide such immunogenic polypeptides in edible plant tissue to elicit an immune response in humans and animals when the plant tissues are consumed. These and other objects of the invention are provided by one or more of the embodiments that are further described in the body of the invention.

Oral Immunology Using Plant Product Containing a Hepatitis Surface Antigen

U.S. Patent: Pending (Allowed)
Application Number: 09/420,695; 09/464,416; 09/464,414

Summary: A method for obtaining an immune response to a non-enteric pathogen antigen (NEPA) such as hepatitis B surface antigen (HBsAg) by feeding the antigen in a plant material to an animal that is immunoreceptive to the NEPA. It has now been discovered that the animal may be made immunoreceptive to the NEPA such as HBsAg by prior primary immunization. When the animal is made immunoreceptive by a prior, e.g. primary, immunization, an immune response to the NEPA may be boosted in the animal by feeding the animal the plant material containing the NEPA. For example, an animal, e.g. a human, that previously had a positive response to primary immunization against hepatitis B, can have a booster response to HBsAg by feeding the animal the antigen in a plant material. The plant material is a substance comprising a physiologically acceptable plant material, especially potatoes, containing the NEPA, e.g. hepatitis B surface antigen (HBsAg). The NEPA, e.g. HBsAg in the plant results from expression by the plant of the NEPA due to genetic alteration.

Detail: Transgenic plants, e.g. potatoes, have been developed that express hepatitis B surface antigen, an antigen known to raise an immune response to hepatitis B when parenterally administered. Unfortunately it has been found that such an immune response is not raised to an acceptable level when the plant, e.g. potato, is simply fed to an animal. It has, however, now been unexpectedly discovered that an immune response to non-enteric pathogen antigens, e.g., hepatitis B surface antigen (HBsAg) may be obtained when the antigen in a plant material is fed to the animal when the animal is immunoreceptive to the HBsAg. It has now been discovered that the animal may be made immunoreceptive to the non-enteric pathogen antigen, e.g. HBsAg, by administering the plant material containing the antigen in conjunction with a suitable adjuvant. The animal may also be immunoreceptive due to a prior, e.g. primary, immunization in which case an immune response to the non-enteric antigen, e.g. HBsAg may be boosted in the animal by feeding the animal the plant material containing the antigen. In such a case it has been found that no adjuvant is needed. An adjuvant may, however, be used with the goal of obtaining even higher immune response. For example, an animal, e.g. a human, that previously had a positive response to primary immunization against hepatitis B, can have a booster response to HBsAg by feeding the animal the antigen in a plant material. The plant material is a substance comprising a physiologically acceptable plant material from a plant (e.g. juice, pulp, leaves, stems, roots, fruit seeds, solids or the whole plant), especially potatoes, containing hepatitis B surface antigen (HBsAg). The HBsAg in the plant results from expression by the plant of HBsAg due to genetic alteration.