Claims
- 1. Lymphotoxin-β, a lymphocyte membrane-type polypeptide comprising SEQ ID NO:2.
- 2. The polypeptide according to claim 1 wherein the polypeptide is associated with a cell surface.
- 3. The polypeptide according to claim 2 wherein the polypeptide is associated with the surface of OKT3-stimulated primary T cells, antigen-specific IL-2 dependent CTL clones, and a PMA-stimulated human T cell hybridoma, II-23.D7.
- 4. A soluble lymphotoxin-13 peptide comprising an amino acid sequence selected from the group consisting of:
(a) SEQ ID NO:4; (b) SEQ ID NO:6; and (c) an amino acid sequence represented by the following formula:
X—SEQ ID NO:6, wherein X comprises one or more of the amino acid residues starting from the 3′ end of SEQ ID NO:8.
- 5. A peptide according to claim 4 further comprising a leader sequence at the 5′ end.
- 6. A polypeptide comprising an amino acid sequence that is encoded by a DNA from the group consisting of:
(a) a DNA sequence comprising SEQ ID NO: 1; (b) DNA sequences that hybridize to the DNA defined by SEQ ID NO:1 and that code on expression for a polypeptide that is substantially homologous with lymphotoxin-β; and (c) DNA comprising degenerate nucleotide sequences that code for the polypeptide that is encoded by the DNA sequence defined by SEQ ID NO:1.
- 7. A polypeptide comprising an amino acid sequence that is encoded by a DNA from the group consisting of:
(a) a DNA sequence comprising SEQ ID NO:3; (b) a DNA sequence comprising SEQ ID NO:5; (c) a DNA sequence represented by the following formula:
X—SEQ ID NO:5, wherein X comprises one or more of the nucleoside triplets starting from the 3′ end of SEQ ID NO:7; (d) DNA sequences that hybridize to any one of SEQ ID NO:3, SEQ ID NO:5 and the sequence according to part (c) above and that code on expression for a polypeptide that is substantially homologous with a soluble lymphotoxin-β peptide; and (e) a DNA sequence comprising degenerate nucleotide sequences that code for the polypeptide encoded for any one of SEQ ID NO:3, SEQ ID NO:5 and the sequence according to part (c) above.
- 8. An engineered polypeptide comprising the amino acid sequence defined by SEQ ID NO:2 wherein the sequence Leu Gly Leu is cleaved from the 5′ end of said sequence and replaced by a single Met or Leu residue.
- 9. An isolated DNA sequence selected from the group consisting of:
(a) a DNA sequence comprising the nucleotide sequence defined by SEQ ID NO: 1; (b) a DNA sequence that hybridizes with the DNA sequence defined by SEQ ID NO:1 and that codes on expression for a polypeptide that is substantially homologous with lymphotoxin-β; and (c) a DNA sequence comprising degenerate nucleotide sequences that code for lymphotoxin-β.
- 10. An isolated DNA sequence selected from the group consisting of:
(a) a DNA sequence comprising the nucleotide sequence defined by SEQ ID NO:3; (b) a DNA sequence comprising the nucleotide sequence defined by SEQ ID NO:5; (c) a DNA sequence comprising the nucleotide sequence according to claim 7(c); (d) DNA sequences that hybridize to a DNA sequence as defined by any one of SEQ ID NO:3, SEQ ID NO:5 or the sequence according to claim 7(c) and that code on expression for a polypeptide that is substantially homologous with a soluble lymphotoxin-β peptide; and (e) a DNA sequence comprising degenerate nucleotide sequences that code for a soluble lymphotoxin-β peptide.
- 11. An engineered DNA sequence comprising the nucleotide sequence defined by SEQ ID NO:1 wherein the nucleotides CTGGGGCTG are cleaved from the 5′ end of said sequence and replaced by a single start codon.
- 12. A recombinant DNA molecule comprising a DNA sequence selected from the group consisting of:
(a) a DNA sequence defined by SEQ ID NO:1; (b) a DNA sequence defined by SEQ ID NO:3; (c) a DNA sequence defined by SEQ ID NO:5; (d) a DNA sequence according to claim 7(c); (e) a DNA sequence according to claim 11;(f) a DNA sequence that hybridizes with the DNA sequences defined by any one of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 and the sequence according to claim 7(c) and that codes on expression for lymphotoxin-β or a soluble lymphotoxin-β peptide; (g) a DNA sequence comprising degenerate nucleotide sequences that codes for lymphotoxin-β; and (h) a DNA sequence comprising degenerate nucleotide sequences that codes for a soluble lymphotoxin-β peptide.
- 13. A host selected from the group consisting of unicellular hosts, animal cells in culture and human cells in culture, transfected with the recombinant DNA molecule of claim 12.
- 14. The host according to claim 13 selected from the group of tumor infiltrating lymphocytes, lymphokine activated killer cells, killer cells and genetically engineered tumor cells removed from a patient.
- 15. A method for producing the polypeptide of any one of claims 1 to 8, said method comprising the steps of culturing a transformed host according to claim 13 and collecting the polypeptide.
- 16. A polypeptide complex comprising a first polypeptide selected from a group consisting of the amino acid sequence defined by any one of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, a polypeptide according to claim 8, and soluble lymphotoxin-β peptide according to claim 4(c), and a second polypeptide selected from the group consisting of lymphotoxin-α, native human or animal lymphotoxin, recombinant lymphotoxin, soluble lymphotoxin, secreted lymphotoxin, or lymphotoxin or lymphotoxin-active fragments of any of the above.
- 17. A polypeptide complex comprising a plurality of lymphotoxin-β polypeptide units.
- 18. A polypeptide complex according to claim 16 wherein the complex is associated with a cell surface.
- 19. A polypeptide complex according to claim 18 wherein the first polypeptide is associated with the surface of OKT3-stimulated primary T cells, antigen-specific IL-2 dependent CTL clones, and a PMA-stimulated non-lymphotoxin human T cell hybridoma, II-23.D7.
- 20. A method for producing lymphotoxin epitopes on the surface of a cell comprising the steps of transfecting the cell with a recombinant DNA molecule according to claim 12 and expressing that DNA in the cell.
- 21. A method for enhancing the targeting tumorcidal activity of tumor infiltrating lymphocytes comprising the steps of transfecting the lymphocytes with a recombinant DNA molecule according to claim 12 and introducing the transformed lymphocytes to a patient.
- 22. The method according to claim 21, wherein the transformed lymphocytes are incubated with a lymphokine before or after transfection with the recombinant DNA molecule according to claim 12.
- 23. The method according to claim 22, wherein the lymphokine is IL-2.
- 24. A composition for preventing, treating or lessening the advancement, severity or effects of HIV infection, neoplasia, inflammation or inflammatory disease, or autoimmune disease comprising an effective amount of a polypeptide selected from the group consisting of a polypeptide according to any one of claims 1 to 8, a polypeptide complex according to any one of claims 16-19, antibodies to any one of the above, or a combination of any of the above, and a pharmaceutically acceptable carrier.
- 25. A method for preventing, treating or lessening the advancement, severity or effects of HIV infection, neoplasia, inflammation or inflammatory diseases, or autoimmune disease comprising administering an effective amount of a polypeptide selected from the group consisting of a polypeptide according to any one of claims 1-8, a polypeptide complex according to any one of claims 16-19, antibodies to any one of the above, or a combination of any of the above, and a pharmaceutically acceptable carrier.
- 26. A composition for suppressing the immune system comprising an effective amount of a polypeptide selected from the group consisting of a polypeptide according to any one of claims 1 to 8, a polypeptide complex according to any one of claims 16-19, antibodies to any one of the above, or a combination of any of the above, and a pharmaceutically acceptable carrier.
- 27. A method for suppressing the immune system comprising administering an effective amount of a polypeptide selected from the group consisting of a polypeptide according to any one of claims 1-8, a polypeptide complex according to any one of claims 16-19, antibodies to any one of the above, or a combination of any of the above, and a pharmaceutically acceptable carrier.
- 28. A nucleotide sequence coding for lymphotoxin-β comprising the nucleotide sequence represented by SEQ ID NO:1 and further comprising an engineered nucleotide sequence at the 5′ end wherein said engineered sequence comprises a functional start codon that is either ATG or CTG and wherein any other codon within said engineered sequence coding for leucine is not CTG.
Parent Case Info
[0001] The present application is a continuation-in-part of co-pending applications PCT/US91/04588, filed Jun. 27, 1991 and U.S. Patent Application. Ser. No. 07/544,862, filed Jun. 27, 1990.
Government Interests
[0002] The invention described herein was made in part during the course of work under Grant No. CA 35638-07-10 from the National Institutes of Health. The U.S. Government has certain rights in this invention.
Divisions (1)
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Number |
Date |
Country |
Parent |
08222614 |
Apr 1994 |
US |
Child |
08466254 |
Jun 1995 |
US |
Continuations (2)
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Number |
Date |
Country |
Parent |
08466254 |
Jun 1995 |
US |
Child |
10040281 |
Nov 2001 |
US |
Parent |
07990304 |
Dec 1992 |
US |
Child |
08222614 |
Apr 1994 |
US |
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
PCT/US91/04588 |
Jun 1991 |
US |
Child |
07990304 |
Dec 1992 |
US |
Parent |
07544862 |
Jun 1990 |
US |
Child |
07990304 |
Dec 1992 |
US |