Dendritic cell vaccines for asparaginyl-β-hydroxylase expressing tumors

• US 9,687,537 B2
• Filed: 07/23/2010
• Issued: 06/27/2017
• Est. Priority Date: 07/24/2009
• Status: Active Grant

• Current Assignee: Rhode Island Hospital
• Original Assignee: Rhode Island Hospital
• Inventors: Shimoda, Masafumi, Wands, Jack R.
• Primary Examiner(s): CANELLA, KAREN A
• Application Number:
  US12/842,494
• Publication Number:
  US 20110076290A1
• Time in Patent Office: 2,531 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 39/4615 Dendritic cells
• CPC Class Codes: A61K 39/4622 Antigen presenting cells
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/24 Interferons [IFN]
• CPC Class Codes: C12N 2501/52 CD40, CD40-ligand (CD154)
• CPC Class Codes: C12N 5/0639 Dendritic cells, e.g. Langherhans cells in the epidermis
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)

Abstract

A vaccine containing AAH-loaded mature dendritic cells for treatment of AAH-expressing tumors in mammalian subjects. A method of producing primed dendritic cells is carried out by contacting isolated dendritic cells with an antigen such as AAH. Following the antigen-contacting step, the dendritic cells are contacted with a combination of cytokines such as GM-CSF and IFN-γ.

Claims

1. A method of reducing growth of an aspartyl (asparaginyl)-β

-hydroxylase (AAH)-expressing tumor in a subject, comprising providing an isolated mature AAH-loaded dendritic cell and administering to said subject said isolated mature AAH-loaded dendritic cell, wherein growth of said AAH-expressing tumor is reduced after administration of said dendritic cell, and wherein said dendritic cell is primed by contact with an AAH followed by contact with a combination of cytokines comprising GM-CSF (Granulocyte-macrophage colony stimulating factor), CD40L (Cluster of Differentiation 40 ligand), and IFN-γ

(interferon gamma) ex vivo, prior to said administering to said subject.

2. The method of claim 1, wherein said tumor is selected from the group consisting of liver, gastrointestinal, pancreas, breast, prostate, cervix, ovary, fallopian tube, larynx, lung, thyroid, gall bladder, kidney, bladder, and brain cancers.

3. The method of claim 1, wherein said dendritic cells are contacted with said combination of cytokines for at least 10 hours.

4. The method of claim 1, wherein said dendritic cells are contacted with said combination of cytokines for at least 40 hours.

5. The method of claim 1, wherein said subject is a human, canine, feline, equine, bovine, or porcine subject.

6. The method of claim 1, wherein said AAH-expressing tumor is a liver cancer.

7. The method of claim 6, wherein said liver cancer comprises hepatocellular carcinoma or cholangiocarcinoma.

8. A method of producing primed dendritic cells, comprising contacting isolated dendritic cells with an antigen comprising AAH (aspartyl (asparaginyl)-β

-hydroxylase), and following said antigen-contacting step, contacting said dendritic cells with a combination of cytokines, said combination comprising GM-CSF (Granulocyte-macrophage colony stimulating factor), CD40L (Cluster of Differentiation 40 ligand), and IFN-γ

(interferon gamma).

9. The method of claim 8, wherein said combination further comprises IL-4 (Interleukin-4).

10. The method of claim 8, wherein said antigen is in a soluble form.

11. The method of claim 8, wherein said dendritic cells are obtained from a subject by leukapheresis.

12. The method of claim 11, wherein said subject is suffering from an AAH-bearing tumor.

13. A vaccine composition comprising primed dendritic cells produced by the method of claim 8.

14. A method of inhibiting tumor growth in a mammal, comprising identifying a subject suffering from an AAH (aspartyl (asparaginyl)-β

-hydroxylase)-bearing tumor, and administering to said subject autologous dendritic cells, wherein said autologous dendritic cells are primed according to the method of claim 8.

15. A method of preventing metastasis of an AAH (aspartyl (asparaginyl)-β

-hydroxylase)-bearing tumor, comprising identifying a subject suffering from an AAH-bearing tumor, and administering to said subject autologous dendritic cells, wherein said autologous dendritic cells are primed according to the method of claim 8.

16. A method of producing primed dendritic cells, comprising contacting isolated dendritic cells with an antigen comprising AAH (aspartyl (asparaginyl)-β

-hydroxylase) and AFP (α

-fetoprotein), and following said antigen-contacting step, contacting said dendritic cells with a combination of cytokines, said combination comprising GM-CSF (Granulocyte-macrophage colony stimulating factor) and IFN-γ

(interferon gamma).

17. A method of producing primed dendritic cells, comprising contacting isolated dendritic cells with an antigen comprising AAH (aspartyl (asparaginyl)-β

-hydroxylase), and following said antigen-contacting step, contacting said dendritic cells with a combination of cytokines, said combination comprising GM-CSF (Granulocyte-macrophage colony stimulating factor), CD40L (Cluster of Differentiation 40 ligand) and IFN-γ

(interferon gamma), and wherein said antigen is bound to a biodegradable solid support.

18. A method of producing primed dendritic cells, comprising contacting isolated dendritic cells with an antigen comprising AAH (aspartyl (asparaginyl)-β

-hydroxylase), and following said antigen-contacting step, contacting said dendritic cells with a combination of cytokines, said combination comprising GM-CSF (Granulocyte-macrophage colony stimulating factor), CD40L (Cluster of Differentiation 40 ligand) and IFN-γ

(interferon gamma), and wherein said antigen is bound to a solid support, wherein said solid support comprises a polystyrene bead.

19. A vaccine containing AAH-loaded mature dendritic cells for treatment of AAH-expressing tumors in a mammalian subject, wherein said dendritic cells are primed by contact with an AAH followed by contact with a combination of cytokines comprising GM-CSF (Granulocyte-macrophage colony stimulating factor), CD40L (Cluster of Differentiation 40 ligand) and IFN-γ

(interferon gamma).

20. A method of reducing growth of an aspartyl (asparaginyl)-β

-hydroxylase (AAH)-expressing tumor in a subject, comprising administering to said subject an isolated mature AAH-loaded dendritic cell, wherein growth of said AAH-expressing tumor is reduced after administration of said dendritic cell, and wherein said dendritic cell is activated with a combination of cytokines comprising GM-CSF and IFN-γ

ex viva, prior to said administering to said subject, wherein said dendritic cell is further loaded with AFP.

21. A method of reducing growth of an aspartyl (asparaginyl)-β

-hydroxylase (AAH)-expressing AAH-expressing liver tumor in a subject, comprising providing an isolated mature AAH-loaded dendritic cell and administering to said subject said isolated mature AAH-loaded dendritic cell, wherein growth of said AAH-expressing tumor is reduced after administration of said dendritic cell, and wherein said dendritic cell is primed by contact with an AAH and a combination of cytokines comprising GM-CSF (Granulocyte-macrophage colony stimulating factor), CD40L (Cluster of Differentiation 40 ligand), interleukin-4 (IL-4) and IFN-γ

(interferon gamma) ex viva, prior to said administering to said subject.

22. The method of claim 21, wherein said AAH-expressing liver tumor comprises hepatocellular carcinoma or cholangiocarcinoma.