Tolerogenic antigen-presenting cells

• US 20040072348A1
• Filed: 03/28/2003
• Published: 04/15/2004
• Est. Priority Date: 03/28/2002
• Status: Abandoned Application

• Current Assignee: Revivicor, Inc (United Therapeutics Corporation)
• Original Assignee: Revivicor, Inc (United Therapeutics Corporation)
• Inventors: Moore, Marilyn Jean, Innes, Donald Alan James, Leishman, Andrew James
• Application Number:
  US10/402,442
• Publication Number:
  US 20040072348A1
• Time in Patent Office: Days
• US Class Current: 435/372
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 39/4615 Dendritic cells
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/4622 Antigen presenting cells
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/464411 Immunoglobulin superfamily
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/064 Immunosuppressive dendritic cells

Abstract

It has been found that dendritic cells can be prepared which cannot mature. These cells can provide signal 1 to T cells but cannot provide co-stimulatory signal 2. T cells which are stimulated by the permanently immature dendritic cells therefore anergise, so the dendritic cells are tolerogenic rather than immunogenic. The cells are generally CD40^−ve, CD80^−ve and CD86^−ve, and remain so when stimulated by inflammatory mediators such as lipopolysaccharide. The cells can be prepared conveniently by the culturing adherent embryonic stem cells in the presence of GM-CSF.

Claims

1. A dendritic cell which is immature and cannot mature.

5. The cell of any one of claims 1 to 4, wherein the cell is not immortal.

6. The cell of any one of claims 1 to 4, wherein the cell has a normal karyotype.

7. The cell of any one of claims 1 to 4, wherein the cell is a human cell.

16. The cells of any one of claims 1 to 4 for use as a medicament.

17. The use of the cells of any one of claims 1 to 4 in the manufacture of a medicament for inhibiting an autoimmune reaction.

18. The use of the cells of any one of claims 1 to 4 in the manufacture of a medicament for inhibiting graft rejection in a recipient.

19. A method of inhibiting graft rejection in a recipient, wherein the cells of any one of claims 1 to 4 are administered to the recipient.

20. A method for transplanting a graft into a recipient, wherein the method also involves the administration of the cells of any one of claims 1 to 4 to the recipient.

21. The method or use of any one of claims 18 to 20, wherein the graft is heart, lung, kidney, liver, pancreas, islets of Langerhans, pancreatic β

-cells or other insulin-producing cells, cornea, bone marrow or nervous tissue.

22. The method or use of any one of claims 18 to 20, wherein the dendritic cells are histocompatible with the graft.

23. A method of inhibiting an autoimmune reaction in a patient, wherein the cells of any one of claims 1 to 4 are administered to the patient.

24. A kit comprising (a) the cells of any one of claims 1 to 4 and (b) a tissue graft for transplanting into a recipient, wherein (a) and (b) are histocompatible.

25. A composition comprising the cells of any one of claims 1 to 4 and a pharmaceutical carrier.

2. A dendritic cell which is able to present antigens to T cells, which is CD40⁻

ve CD80<sup>−

ve</sup> and CD86<sup>−

ve</sup>, and which remains CD40<sup>−

ve</sup> CD80<sup>−

ve</sup> and CD86<sup>−

ve</sup> when stimulated by inflammatory mediators.

3. A dendritic cell which can deliver signal 1 to a T cell, but which cannot provide signal 2 to the T cell, either in a resting state or when stimulated by an inflammatory mediator.

4. A tolerogenic dendritic cell differentiated in vitro from an ES cell.

9. A process for preparing a tolerogenic antigen-presenting cell from a stem cell, wherein the method includes the step of culturing the stem cell in the presence of one or more cytokine(s) which cause(s) the stem cell to differentiate into the tolerogenic cell.

8. A cell obtainable by the method of any one of claims 9 to 15.

10. The process of claim 9, wherein the stem cell is an embryonic stem cell.

11. The process of claim 9 or claim 10, wherein the stem cell is a human stem cell.

12. The process of any one of claims 9 to 11, wherein the stem cells develop into embryoid bodies before differentiation into the tolerogenic cells.

13. The process of any one of claims 9 to 12, wherein differentiation into the tolerogenic cell takes place in adherent culture.

14. The process of any one of claims 9 to 13, wherein a feeder layer is not used.

15. The process of any one of claims 9 to 14, wherein the cytokine is GM-CSF.

26. A stem cell for use in the process of any one of claims 9 to 15, wherein the stem cell has been genetically manipulated.

27. The stem cell of claim 26, wherein the stem cell has been genetically manipulated to encode a polypeptide which promotes differentiation of the stem cell into a dendritic cell.

28. The stem cell of claim 26, wherein the stem cell has been genetically manipulated to express or over-express one or more surface proteins which down-regulate immune responses.

29. The stem cell of claim 26, wherein the stem cell has been genetically manipulated not to express or to under-express surface and/or secreted proteins which promote T cell activation.

30. The stem cell of claim 26, wherein the stem cell has been genetically manipulated to include a suicide gene.

31. The stem cell of claim 26, wherein the stem cell has been genetically manipulated to include a marker suitable for lineage selection.