Methods and compositions for protection against lentiviral infections
View Patent ↗The present invention provides methods and compositions for optimally co-expressing in a primate subject a tyrosylprotein sulfotransferase (TPST) and a lentiviral gp120-binding molecule to provide potent and long term protection against lentiviral infections.
1. A method of preventing a lentiviral viral infection in a subject at risk of developing or suffering from the lentiviral infection, comprising administering to the subject a prophylactically effective amount of:
(a) a first polynucleotide sequence expressing a tyrosylprotein sulfotransferase (TPST), and
(b) a second polynucleotide sequence expressing a fusion polypeptide comprising:
(i) a CCR5 mimetic comprising SEQ ID NO: 21 that binds a gp120 protein of a primate lentivirus,
(ii) a CD4 polypeptide having at least 90% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 4, and
(iii) an immunoglobulin Fc domain;
wherein the molar ratio of the first polynucleotide sequence to the second polynucleotide sequence, or the molar ratio of the TPST expressed from the first polynucleotide sequence to the fusion polypeptide expressed from the second polynucleotide sequence, is at least 1:2; thereby preventing the viral infection in the subject.
2. The method of claim 1 , wherein the first polynucleotide sequence and the second polynucleotide sequence are provided in a therapeutic composition.
3. The method of claim 1 , wherein the molar ratio is at least 1:1.
4. The method of claim 1 , wherein the first polynucleotide sequence and the second polynucleotide sequence are each present in an expression vector.
5. The method of claim 4 , wherein each of the expression vectors is independently an adeno-associated virus (AAV) vector, an adenovirus vector, a retrovirus vector, a lentivirus vector, or a herpesvirus vector.
6. The method of claim 1 , wherein the first polynucleotide sequence and the second polynucleotide sequence are both present in one expression vector.
7. The method of claim 6 , wherein the expression vector is an adeno-associated virus (AAV) vector, an adenovirus vector, a retrovirus vector, a lentivirus vector, or a herpesvirus vector.
8. The method of claim 1 , wherein the TPST is tyrosylprotein sulfotransferase 2 (TPST2) or an enzymatically active fragment thereof.
9. The method of claim 1 , wherein the fusion polypeptide binds to a coreceptor binding site on the gp120 protein.
10. The method of claim 1 , wherein the fusion polypeptide comprises an amino acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID Nos: 9-11 and 16-19.
11. The method of claim 10 , wherein the fusion polypeptide is eCD4-Ig having an amino acid sequence shown in SEQ ID NO: 11.
12. The method of claim 6 , wherein the expression vector comprises a polynucleotide sequence that expresses a first polypeptide and a second polypeptide at a molar ratio of at least 1:2, wherein the first polypeptide is a tyrosine-protein sulfotransferase (TPST) 1 or TPST2, and the second polypeptide is a fusion protein comprising (i) a CCR5 mimetic comprising SEQ ID NO: 21 that binds a gp120 protein of a primate lentivirus, (ii) a CD4 polypeptide having at least 90% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 4, and (iii) an immunoglobulin Fc domain.
13. The method of claim 6 , wherein the expression vector further comprises one or more expression regulatory elements to control the molar ratio, wherein the regulatory elements are selected from the group consisting of an enhancer element, an intron, an internal ribosome entry site (IRES), and a woodchuck response element (WPRE).
14. The method of claim 1 , wherein the subject is a human, and the wherein the lentiviral infection is HIV-1 infection.
15. The method of claim 1 , wherein the TPST is TPST2, and the fusion polypeptide comprises eCD4-Ig (SEQ ID NO:11), eCD4-Ig mim2 (SEQ ID NO: 16), eCD4-Ig Q40A (SEQ ID NO:17), or eCD4-Ig Q40A,mim2 (SEQ ID NO:18).