Porcine hepatocytes for use in treatment of disorders characterized by insufficient liver function

• US 6,610,288 B1
• Filed: 05/24/1996
• Issued: 08/26/2003
• Est. Priority Date: 05/26/1995
• Status: Expired due to Fees

• Current Assignee: Boston Medical Center, GenVec, Inc. (Precigen, Inc.), Trustees of Boston University
• Original Assignee: Diacrin, Inc. (Precigen, Inc.), Trustees of Boston University
• Inventors: Gunsalus, J. Ryan, Afdhal, Nezam H., Edge, Albert
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US08/653,059
• Time in Patent Office: 2,650 Days
• Field of Search: 435/240.2, 435/325, 424/93.1, 424/93.21, 424/93.3, 424/139.1, 424/184.1, 530/387.1, 530/388.85, 600/19
• US Class Current: 424/93.2
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/407 Liver; Hepatocytes
• CPC Class Codes: C07K 16/2833 against MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C12N 5/067 Hepatocytes
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Isolated porcine hepatocytes, isolated populations of such hepatocytes and methods for using the hepatocytes to treat subjects with disorders characterized by insufficient liver function are described. The porcine hepatocytes can be either hepatocytes isolated from adult, immature, or embryonic swine. The porcine hepatocytes can be modified to be suitable for transplantation into a xenogeneic subject, for example, by altering an antigen (e.g., an MHC class I antigen) on the cell surface which is capable of stimulating an immune response against the cell in the subject (e.g., by contact with an anti-MHC class I antibody, or a fragment or derivative thereof). The isolated porcine hepatocytes of the invention can be used to treat disorders characterized by insufficient liver function by administering the hepatocytes to a subject having such a disorder.

Claims

1. A method of treating a xenogeneic subject suffering from a disorder characterized by decreased liver function, comprising

i) isolating a population of porcine hepatocytes

ii) altering the hepatocytes by contacting the hepatocytes prior to introduction into a subject with at least one anti-MAC class I antibody, which binds to the MHC class I antigen on the cell surface but does not activate complement or induce lysis of the cells such that rejection of the hepatocytes is inhibited upon introduction of the cells into the xenogeneic subject;

and

iii) administering the porcine hepatocytes to the liver of the subject while decreasing portal blood pressure of the subject to thereby treat a xenogeneic subject suffering from a disorder characterized by decreased liver function.

2. The method of claim 1, wherein the porcine hepatocytes are obtained from an embryonic pig.

3. The method of claim 1, wherein the subject is a human.

4. The method of claim 1, wherein the disorder is a genetic disease involving a liver enzyme.

5. The method of claim 4, wherein the genetic disease is Crigler-Najjar Syndrome Type I or ornithine transcarbamoylase deficiency.

6. The method of claim 1, wherein the disorder is a genetic disease involving a protein secreted by the liver.

7. The method of claim 6, wherein the genetic disease is hereditary emphysema or hemophilia.

8. The method of claim 1, wherein the disorder is acute liver failure.

9. The method of claim 1, wherein the disorder is chronic liver failure.

10. The method of claim 1, wherein the disorder is hepatitis A, hepatitis B, or non-A, non-B hepatitis.

11. The method of claim 1, wherein the step of decreasing portal blood pressure comprises using a transjugular intra-hepatic porto-systemic shunt to decrease portal blood pressure.

12. The method of claim 1, further comprising administering an immunosuppressive agent to the subject.

13. The method of claim 1, wherein the porcine hepatocytes are free from at least one organism selected from the group consisting of parasites, bacteria, mycoplasma, and viruses.

14. The method of claim 1, wherein the disorder is familial hypercholesterolemia.

15. A method of treating a xenogeneic subject suffering from a disorder characterized by decreased liver function, comprising

i) isolating a population of porcine hepatocytes

ii) altering said porcine hepatocytes by contacting the hepatocytes prior to introduction into a subject with monoclonal antibody PT85, which binds to the MHC class I antigen on the cell surface but does not activate complement or induce lysis of the hepatocytes such that rejection of the cells is inhibited upon introduction of the hepatocytes into the xenogeneic subject;

and

iii) administering the porcine hepatocytes to the liver of the subject while decreasing portal blood pressure of the subject to thereby treat a xenogeneic subject suffering from a disorder characterized by decreased liver function.

16. A method of treating a xenogeneic subject suffering from a disorder characterized by decreased liver function, comprising

i) isolating a population of porcine hepatocytes

ii) altering the porcine hepatocytes by contacting the hepatocytes prior to introduction into a subject with at least one F(ab′

)2 fragment of an anti-MHC class I antibody, which binds to the MHC class I antigen on the cell surface but does not activate complement or induce lysis of the hepatocytes such that rejection of the hepatocytes is inhibited upon introduction of the hepatocytes into the xenogeneic subject;

iii) administering the porcine hepatocytes to the liver of the subject while decreasing portal blood pressure of the subject to thereby treat a xenogeneic subject suffering from a disorder characterized by decreased liver function.

17. The method of claim 16, wherein the porcine hepatocytes are obtained from an embryonic pig.

18. The method of claim 16, wherein the subject is a human.

19. The method of claim 16, wherein the disorder is a genetic disease involving a liver enzyme.

20. The method of claim 16, wherein the genetic disease is Crigler-Najjar Syndrome Type I or ornithine transcarbamoylase deficiency.

21. The method of claim 16, wherein the disorder is a genetic disease involving a protein secreted by the liver.

22. The method of claim 16, wherein the genetic disease is hereditary emphysema or hemophilia.

23. The method of claim 16, wherein the disorder is acute liver failure.

24. The method of claim 16, wherein the disorder is chronic liver failure.

25. The method of claim 16, wherein the disorder is hepatitis A, hepatitis B, or non-A, non-B hepatitis.

26. The method of claim 16, wherein the disorder is familial hypercholesterolemia.

27. The method of claim 16, wherein the step of decreasing portal blood pressure comprises using a transjugular intra-hepatic porto-systemic shunt to decrease portal blood pressure.

28. The method of claim 16, further comprising administering an immunosuppressive agent to the subject.

29. The method of claim 16, wherein the porcine hepatocytes are free from at least one organism selected from the group consisting of parasites, bacteria, mycoplasma, and viruses.