Compositions and methods for self-adjuvanting vaccines against microbes and tumors

Patent metadata

Field	Value
Publication number	US-10238734-B2
Application number	US-201113635885-A
Country	US
Kind code	B2
Filing date	Mar 22, 2011
Priority date	Mar 23, 2010
Publication date	Mar 26, 2019
Grant date	Mar 26, 2019

Abstract

The present invention is drawn to compositions and methods to enhance an immune response in order to prevent or treat infections or hyperproliferative diseases such as cancer. More particularly, the composition is an immunostimulatory intracellular signaling peptide fused directly or indirectly to a peptide that leads to multimerization into complexes of three or more units, where the intracelluar signaling peptide must be present in a complex of three or more units in order to stimulate an immune response. Inserting this fusion construct into viruses like HIV-1 or introducing it into dendritic cells or tumor cells is predicted to lead to a positive therapeutic effect in humans, non-human mammals, birds, and fish.

First claim

What is claimed is:

1. An isolated protein comprising a multimerizing domain from the N-terminal fragment of Latent Membrane Protein 1 (LMP1) operatively joined to a cytoplasmic signaling domain of a heterologous receptor such that the protein assembles into a complex of three or more protein moieties; wherein the LMP1 multimerizing domain is translated from nucleic acids that do not contain introns.

2. The protein of claim 1 , wherein the cytoplasmic signaling domain comprises a cytoplasmic fragment of a member of Tumor Necrosis Factor Receptor Superfamily (TNFRSF).

3. The protein of claim 1 , wherein the cytoplasmic signaling domain comprises a cytoplasmic fragment selected from the group consisting of CD40, CD27, Fas, Lymphotoxin beta receptor (LTBR), nerve growth factor receptor (NGFR), Tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), Tumor necrosis factor receptor superfamily member 1B (TNFRSF1B), Tumor necrosis factor receptor superfamily member 4 (TNFRSF4), Tumor necrosis factor receptor superfamily member 8 (TNFRSF8), Tumor necrosis factor receptor superfamily member 9 (TNFRSF9), Tumor necrosis factor receptor superfamily member 10A (TNFRSF10A), Tumor necrosis factor receptor superfamily member 10B (TNFRSF10B), Tumor necrosis factor receptor superfamily member 10D (TNFRSF10D), Tumor necrosis factor receptor superfamily member 11A (TNFRSF11A), Tumor necrosis factor receptor superfamily member 12A (TNFRSF12A), Tumor necrosis factor receptor superfamily member 13B (TNFRSF13B), Tumor necrosis factor receptor superfamily member 13C (TNFRSF13C), Tumor necrosis factor receptor superfamily member 14 (TNFRSF14), Tumor necrosis factor receptor superfamily member 17 (TNFRSF17), Tumor necrosis factor receptor superfamily member 18 (TNFRSF 18), Tumor necrosis factor receptor superfamily member 19 (TNFRSF19), Tumor necrosis factor receptor superfamily member 21 (TNFRSF21), and Tumor necrosis factor receptor superfamily member 25 (TNFRSF25).

4. The protein of claim 1 , wherein the cytoplasmic signaling domain comprises CD40.

5. The protein of claim 1 , wherein the multimerizing domain comprises at least four transmembrane regions.

6. The protein of claim 1 , comprising a Latent Membrane Protein 1 (LMP 1)-CD40 fusion protein.

7. An isolated virus, microbe, or host cell, comprising:
(i) a first expression cassette for expressing a protein, comprising a multimerizing domain from the N-terminal fragment of Latent Membrane Protein 1 (LMP1); operatively joined to a cytoplasmic signaling domain of a receptor such that the protein assembles into a complex of three or more protein moieties, and wherein, when the protein is Latent Membrane Protein-1 (LMP1), then the nucleic acid encoding LMP1 does not contain introns; and
(ii) a second expression cassette for expressing a protein antigen, with the proviso that the protein antigen is not an Epstein-Barr virus (EBV) protein.

8. The isolated virus, microbe, or host cell of claim 7 , whereby the protein comprises cell stimulatory activity from the cytoplasmic signaling domain.

9. The isolated virus, microbe, or host cell of claim 7 , wherein the protein comprises:
a multimerizing domain comprising an N-terminus fragment of latent membrane protein 1 (LMP1); and
a cytoplasmic signaling domain comprising a cytoplasmic fragment of a member of Tumor necrosis factor receptor superfamily (TNFRSF).

10. The isolated virus, microbe, or host cell of claim 9 , wherein the protein comprises a LMP1-CD40 fusion protein.

11. The isolated virus of claim 7 , wherein the virus expresses the protein of claim 8 (i) after infection or transduction of the virus into cells.

12. The isolated virus or microbe of claim 7 , wherein the virus or microbe expresses the protein of claim 7 (i) after infection or transduction of the virus or microbe into cells, wherein the virus or microbe is selected from the group consisting of Human immunodeficiency virus-1 (HIV-1), Simian immunodeficiency virus (SIV), influenza virus, parainfluenza virus, dengue virus, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Cytomegalovirus (CMV), adenovirus, adeno-associated virus, Simian virus 40 (SV 40), Modified Vaccinia Ankara (MVA), Vesicular stomatitis virus (VSV), arenaviruses, bunyaviruses, flaviviruses, West Nile virus, Japanese Encephalitis virus, Venezuelan equine encephalitis virus, Eastern equine encephalitis virus, Western equine encephalitis, herpesviruses, measles virus, rhabdoviruses, Listeria, Salmonella , and combinations thereof.

13. The isolated virus, microbe, or host cell of claim 7 , wherein the antigen comprises a tumor antigen, viral antigen, or microbial antigen.

14. The isolated host cell of claim 7 , wherein the host cell is a microorganism.

15. An immunogenic composition, comprising:
(i) a delivery system comprising a microorganism other than Epstein-Barr virus (EBV);
(ii) an antigen comprising a tumor antigen, viral antigen, or microbial antigen, wherein the antigen is endogenous to the microorganism used to deliver the immunogenic composition or not normally expressed by the microorganism used to deliver the immunogenic composition, and wherein the antigen is not an EBV protein;
(iii) a protein encoded by the expression cassette of claim 7 (i) and
(iv) wherein the immunogenic composition induces a dendritic cell, macrophage, T cell lymphocyte, or anti-cancer response greater than that induced by the same composition lacking the expression cassette of claim 7 (i).

16. The immunogenic composition of claim 15 , wherein the protein encoded by the expression cassette of claim 7 (i) is comprised of a multimerizing domain comprising an N-terminus fragment of latent membrane protein 1 (LMP1) and a cytoplasmic signaling domain comprising a cytoplasmic fragment of a member of the Tumor necrosis factor receptor superfamily (TNFRSF).

17. The immunogenic composition of claim 15 , wherein the protein encoded by the expression cassette of claim 7 (i) comprises a LMP1-CD40 fusion protein.

18. The immunogenic composition of claim 15 , wherein the delivery system comprises a DNA virus, RNA virus, or prokaryotic organism.

19. The immunogenic composition of claim 15 , wherein the delivery system comprises a virus or microbe selected from the group consisting of Human immunodeficiency virus-I (HIV-1), Simian immunodeficiency virus (SIV), influenza virus, parainfluenza virus, dengue virus, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Cytomegalovirus (CMV), adenovirus, adeno-associated virus, Simian virus 40 (SV 40), Vaccinia virus, Modified Vaccinia Ankara (MVA), Vesicular stomatitis virus (VSV), arenaviruses, bunyaviruses, flaviviruses, West Nile virus, Japanese encephalitis virus, Venezuelan encephalitis virus, Eastern equine encephalitis virus, herpesviruses, measles virus, rhabdovirus, Listeria, Salmonella , and combinations thereof.

20. A method for stimulating an immune response in a subject comprising: administering to a cell an effective amount of a polynucleotide comprising a first expression cassette for expressing the protein of claim 1 , and/or a second expression cassette for expressing an antigen, thereby stimulating an immune response.

21. The method of claim 20 , wherein at least two nucleic acid sequences are administered, where a first nucleic acid sequence comprises the first expression cassette for expressing the protein of claim 1 , and a second nucleic acid sequence comprises the second expression cassette for expressing an antigen, and the two nuc

Assignees

• Kornbluth Richard Syd
• Stone Geoffrey William
• Univ California

Inventors

• Kornbluth Richard Syd
• Stone Geoffrey William

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• C12N2740/15011

  Lentivirus, not HIV, e.g. FIV, SIV · CPC title

• A61K2039/5256

  expressing foreign proteins · CPC title