NOVEL PD1 ISOFORMS, AND USES THEREOF FOR POTENTIATING IMMUNE RESPONSES

• US 20140302070A1
• Filed: 04/07/2014
• Published: 10/09/2014
• Est. Priority Date: 04/05/2013
• Status: Active Grant

• Current Assignee: Versitech Limited (Government of Hong Kong Special Administrative Region)
• Original Assignee: University of Hong Kong (Government of Hong Kong Special Administrative Region)
• Inventors: Zhou, Jingying, Chen, Zhiwei, Cheng, Lin, Cheung, Allen Ka Loon
• Granted Patent: US 10,047,137 B2
• Time in Patent Office: Days
• US Class Current: 424/185.1
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/55516 Proteins; Peptides
• CPC Class Codes: A61K 39/0008 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: C07K 14/70503 Immunoglobulin superfamily
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C12N 2740/16234 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Abstract

In one embodiment, the present invention provides a new isoform of human PD1 (Δ42PD1) that contains a 42-nucleotide in-frame deletion located at exon 2 domain. Δ42PD1 does not engage PD-L1/PD-L2, and can induce the production of pro-inflammatory cytokines In one embodiment, Δ42PD1 can be used as an intramolecular adjuvant to develop a fusion DNA vaccine for enhancing antigen-specific CD8⁺ T cell immunity and for prevention of pathogenic infection and/or cancer. In one embodiment, soluble Δ42PD1 protein could be a therapeutic target for autoimmune diseases. In other embodiments, proteins or peptides or nucleic acids encoding proteins or peptides containing Δ42PD1 could be used as immunogens for developing antibodies binding specifically to Δ42PD1. In yet another embodiment, neutralizing antibodies could block sΔ42PD1 function and accordingly could be used as treatment for autoimmune disorders.

Claims

1. An isolated PD1 protein isoform having a deletion of 14 amino acids at positions corresponding to amino acids 26-39 of the wild-type PD1 protein having an amino acid sequence of SEQ ID NO:

2, or a fragment of the PD1 protein isoform, wherein the fragment has at least 30 amino acids and wherein the fragment has said deletion of 14 amino acids at positions corresponding to amino acids 26-39 of the wild-type PD1 protein having an amino acid sequence of SEQ ID NO;

2.

2. The PD1 protein isoform of claim 1, which does not bind to PDLL or PDL2.

3. The PD1 protein isoform of claim 1, wherein the 14 amino acids deleted from the wild-type PD1 protein are DSPDRPWNPPTFFP (SEQ ID NO:

3).

4. The PD1 protein isoform of claim 1, comprising SEQ ID NO:

1 or a fragment thereof comprising at least 30 consecutive amino acids of SEQ ID NO;

1.

7. An antibody that binds specifically to the PD1 protein isoform of claim 1.

8. The antibody of claim 7, where the antibody cannot bind to wild type PD1.

20. An immunogen that generates an antibody of claim 7.

5. A nucleic acid molecule encoding:

a PD1 protein isoform having a deletion of 14 amino acids at positions corresponding to amino acids 26-39 of the wild-type PD1 protein having an amino acid sequence of SEQ ID NO;

2;

or

a fragment of the PD1 protein isoform wherein the fragment has at least 30 amino acids, wherein the fragment has said deletion of 14 amino acids at positions corresponding to amino acids 26-39 of the wild-type PD1 protein having an amino acid sequence of SEQ ID NO;

2.

6. The nucleic acid molecule of claim 5, wherein the nucleic acid molecule is a cDNA molecule.

11. A fusion nucleic acid molecule comprising a nucleic acid molecule of claim 5 and a nucleic acid molecule encoding an antigen or peptide.

12. A vaccine composition comprising a nucleic acid molecule of claim 5.

13. A vaccine composition comprising a fusion nucleic acid molecule of claim 11.

14. A method of inducing the production of TNF-α

, IL-1, or IL-6, comprising administering to an immune cell a nucleic acid molecule of claim 5.

15. A method of inducing the production of TNF-α

, IL-1, or IL-6, comprising administering to an immune cell a fusion nucleic acid molecule of claim 11.

17. A method of preventing pathogenic infection or cancer, comprising administering to a subject in need of such prevention a nucleic acid molecule of claim 5.

18. A method of preventing pathogenic infection or cancer, comprising administering to a subject in need of such prevention a fusion nucleic acid molecule of claim 11.

9. A fusion protein comprising:

a PD1 protein isoform having a deletion of 14 amino acids at positions corresponding to amino acids 26-39 of the wild-type PD1 protein having an amino acid sequence of SEQ ID NO;

2;

or

a fragment of the PD1 protein isoform wherein the fragment has at least 30 amino acids, wherein the fragment has said deletion of 14 amino acids at positions corresponding to amino acids 26-39 of the wild-type PD1 protein having an amino acid sequence of SEQ ID NO;

2.

10. The fusion protein of claim 9, further comprising an antigen protein or peptide.

16. A method of inducing the production of TNF-α

, IL-1, or IL-6, comprising administering to an immune cell a fusion protein of claim 9.

19. A method of preventing pathogenic infection or cancer, comprising administering to a subject in need of such prevention a fusion protein of claim 9.