Claims
- 1. A purified preparation of immunologically privileged cells which express at least one protein not normally expressed by said cells.
- 2. The cells of claim 1, which are intermediate lobe pituitary cells.
- 3. The cells of claim 1, wherein said protein is expressed from a heterologous nucleic acid sequence inserted into the cell.
- 4. The cells of claim 1, wherein expression of said protein is operatively linked to a control region other than the one which normally controls it.
- 5. The cells of claim 2, wherein expression of said protein is operatively linked to a control region which is active in or specific for the intermediate lobe pituitary cell.
- 6. The cells of claim 2, wherein said protein is from a first species and the intermediate lobe pituitary cells is from a second species.
- 7. Intermediate lobe pituitary cells which express insulin from an insulin-encoding nucleic acid sequence.
- 8. The cells of claim 7, wherein the insulin-encoding nucleic acid sequence is operatively linked to a control region other than the insulin control region.
- 9. The cells of claim 8, wherein said insulin-encoding nucleic acid sequence encodes human insulin.
- 10. The cells of claim 7, wherein said intermediate lobe pituitary cells are chosen from: fetal or non-fetal, human or non-human cells.
- 11. The cells of claim 7, wherein the insulin-encoding nucleic acid encodes human insulin and the intermediate lobe pituitary cells are from another species.
- 12. The cells of claim 7, wherein said cells further express another protein which is not normally expressed in intermediate lobe pituitary cells, and which mediates glucose stimulated insulin secretion.
- 13. The cells of claim 12, wherein said protein promotes the transport of glucose across the plasma membrane such that insulin release by the cells is modulated by glucose levels.
- 14. The cells of claim 12, wherein said cells express glucokinase with a high Km for glucose.
- 15. The cells of claim 14, wherein said cells further express an ion channel which mediates the glucose stimulated insulin secretion.
- 16. The cells of claim 12, wherein said cells further express GLP-1.
- 17. The cells of claim 7 which are encapsulated within a non-antigenic compound.
- 18. The cells of claim 17 wherein the compound is a polymer.
- 19. The cells of claim 17 wherein said compound is hydrogel, olginate or semipermeable fiber.
- 20. Immunologically privileged cells which express human insulin, wherein said cells include:
an insulin-encoding nucleic acid sequence operatively linked to a control region which allows expression in said cells; a nucleic acid sequence which encodes GLUT-2 operatively linked to a control region which allows expression in said cells; a nucleic acid sequence which encodes the β-cell isoform of glucokinase operatively linked to a control region which allows expression in said cells; and a nucleic acid sequence which encodes an ion channel which mediates the expression of insulin operatively linked to a control region which allows expression in said cells.
- 21. The cells of claim 20, further including a nucleic acid which encodes GLP-1.
- 22. The cells of claim 20 which are intermediate lobe pituitary cells.
- 23. Intermediate lobe pituitary cells which express human insulin, wherein said cells include:
a heterologous insulin-encoding nucleic acid sequence operatively linked to a control region other than the insulin control region; a nucleic acid sequence which encodes GLUT-2 operatively linked to a control region other than the GLUT-2 control region; a nucleic acid sequence which encodes the β-cell isoform of glucokinase operatively linked to a control region other than the glucokinase control region; and a nucleic acid sequence which encodes an ion channel which mediates glucose-sensitive insulin secretion operatively linked to a control region which allows expression in intermediate lobe pituitary cells.
- 24. Intermediate lobe pituitary cells which express human insulin, wherein said cells include:
a insulin-encoding nucleic acid sequence operatively linked to a control region which allows expression in intermediate lobe pituitary cells; a nucleic acid sequence which encodes GLUT-2 operatively linked to a control region which allows expression in intermediate lobe pituitary cells; a nucleic acid sequence which encodes the β-cell isoform of glucokinase operatively linked to a control region which allows expression in intermediate lobe pituitary cells; and a nucleic acid sequence which encodes a K+/ATP ion channel which mediates the expression of insulin operatively linked to a control region which allows expression in intermediate lobe pituitary cells.
- 25. The cells of claim 24, further including a nucleic acid which encodes GLP-1 operatively linked to a control region which allows expression in intermediate lobe pituitary cells.
- 26. A method of producing a protein in a subject in vivo comprising introducing into the subject an immunologically privileged cell which expresses the protein.
- 27. The method of claim 26, wherein said cell is an intermediate lobe pituitary cell and the protein is one which is not normally expressed therein.
- 28. The method of claim 27, wherein said intermediate lobe pituitary cells includes a nucleic acid sequence which encodes a protein not normally expressed by an intermediate lobe pituitary cell operatively linked to a heterologous control region which controls expression of the nucleic acid in the intermediate lobe pituitary cell.
- 29. The method of claim 28, wherein said protein is insulin.
- 30. The method of claim 27, wherein said intermediate lobe pituitary cell is an autologous cell, an allogenic cell, or a xenogenic cell.
- 31. The method of claim 28, wherein said subject is a human and the intermediate lobe pituitary cell is an autologous cell, an allogenic cell or a xenogenic cell.
- 32. A transgenic animal expressing at least one protein not normally expressed in intermediate lobe pituitary cells.
- 33. The transgenic animal of claim 32 which is a pig.
- 34. The transgenic animal of claim 33 wherein the protein is insulin.
- 35. The transgenic animal of claim 34 wherein the expression of insulin is controlled in a glucose stimulated insulin secreting manner.
- 36. A subject which has an intermediate lobe pituitary cell which express a peptide not normally expressed in intermediate lobe pituitary cells.
- 37. The subject of claim 36, wherein the cell expresses insulin.
- 38. The subject of claim 37, the expression of insulin is controlled in a glucose stimulated insulin secreting manner.
- 39. A method of treating a subject in need of a protein, comprising introducing immunologically privileged cells which express the protein systemically.
- 40. The method of claim 39 wherein the cells are intermediate lobe pituitary cells and the protein is insulin.
- 41. The method of claim 40 wherein the expression of insulin is controlled in a glucose stimulated insulin secreting manner.
- 42. A method of treating an individual suffering from diabetes or insulin deficiency comprising introducing intermediate lobe pituitary cells which express insulin in a glucose stimulated insulin secreting manner systemically.
- 43. The method of claim 42 wherein the cells further express glucokinase, GLUT-2 and a K+/ATP ion channel.
- 44. The method of claim 42 wherein the cells further express GLP-1.
- 45. A method of expressing a protein not normally expressed in intermediate lobe pituitary cells comprising transfecting, in vivo, said cells with a nucleotide sequence encoding the protein.
- 46. The method of claim 45, wherein the protein is insulin.
- 47. The method of claim 46, further including transfecting said cells with additional nucleotide sequences encoding one or more additional proteins which control expression of insulin in a glucose stimulated insulin secreting manner.
- 48. The method of claim 47, wherein the additional nucleotide sequences encode the following proteins: the β-cell isoform of glucokinase, GLUT-2, and a K+ATP ion channel.
- 49. The method of claim 47, wherein the additional nucleotide sequences encode GLP-1.
- 50. Intermediate lobe pituitary cells which have been transfected to express insulin, and which have further been transfected with nucleotide sequences encoding other proteins, comprising nucleotide sequences encoding:
the β-cell isoform of glucokinase operatively linked to a control region to allow expression in such cells; GLUT-2 operatively linked to a control region to allow expression in such cells; and A K+/ATP ion channel which mediates expression of insulin operatively linked to a control region to allow expression in such cells.
- 51. The method of claim 47, wherein the nucleotide sequences also encode GLP-1.
- 52. A method of expressing a protein in immunologically privileged cells not normally expressing said protein comprising transfecting, in vivo, said cells with a nucleotide sequence encoding said protein.
- 53. The method of claim 52, wherein the protein is insulin.
- 54. The method of claim 53, further including transfecting said cells with additional nucleotide sequences encoding one or more additional proteins which control expression of insulin in a glucose stimulated insulin secreting manner.
- 55. The method of claim 53, wherein the additional nucleotide sequences encode the β-cell isoform of glucokinase, GLUT-2 and a K+/ATP ion channel.
- 56. The method of claim 47, wherein the additional nucleotide sequences also encode GLP-1.
- 57. A method of expressing insulin in cells which do not normally express insulin comprising transfecting, in vivo, said cells with a nucleotide sequence encoding insulin and with additional nucleotide sequences encoding one or more additional proteins which control expression of insulin in a glucose stimulated insulin secreting manner.
- 58. The method of claim 57 wherein the additional nucleotide sequences encode the β-cell isoform of glucokinase, GLUT-2 and a K+/ATP ion channel.
- 59. The method of claim 47, wherein the additional nucleotide sequences also encode GLP-1.
Parent Case Info
[0001] This is a continuation-in-part of U.S. Provisional application Serial No. 60/054,730, filed Aug. 5, 1997, which is incorporated by reference.
Divisions (1)
|
Number |
Date |
Country |
Parent |
09127296 |
Jul 1998 |
US |
Child |
09770601 |
Jan 2001 |
US |