Immunizing compositions comprising HIV-1 proviral constructs with an inactive p6 gag TSG101 UEV binding domain capable of producing budding-defective viral particles that remain tethered to the cell surface
The present invention provides methods for preventing and treating HIV infection and AIDS by introducing cells displaying HIV late-domain phenotype into a patient, or by administering to a patient nucleic acids, polypeptides or small organic compounds to cause the formation of cells displaying HIV late-domain phenotype in the body of the patient.
1. An immunizing composition comprising:
a mutant human immunodeficiency virus 1 (HIV1) proviral construct comprising a deletion or inactivation of the Tsg101 UEV binding domain (P(T/S)AP motif (SEQ ID NO:1 or SEQ ID NO:2) and a pharmaceutically acceptable adjuvant, wherein the expression of said construct in a host cell results in the assembly of HIV1 virus particles that are incapable of budding from the surface of said host cell, and remain attached to the surface of said host cell.
2. The composition of claim 1 , wherein said adjuvant is capable of stimulating a cytotoxic T lymphocyte (CTL) response.
3. The composition of claim 2 , wherein said adjuvant is a cytokine protein or a nucleic acid encoding a cytokine protein.
4. The composition of claim 3 , wherein the cytokine protein is selected from the group consisting of IL-2, IL-4, IL-12, IL-18, gamma-interferon, and a fusion protein thereof.
5. The composition of claim 1 , wherein said construct encodes a mutant HIV Pr55Gag polypeptide devoid of late domain motifs.
6. The composition of claim 5 , wherein said construct is devoid of late domain motifs.
7. The composition of claim 5 , wherein said construct further encodes another HIV protein or an immunogenic fragment thereof.
8. The composition of claim 7 , wherein said HIV protein is a surface protein of HIV viral particles.
9. The composition of claim 8 , wherein said HIV protein is an envelope protein.
10. The composition of claim 1 , wherein said construct lacks the HIV Gag-p6 sequence.
11. The composition of claim 10 , wherein the adjuvant is capable of stimulating a cytotoxic T lymphocyte (CTL) response.
12. The composition of claim 11 , wherein said adjuvant is a cytokine protein or a nucleic acid encoding a cytokine protein.
13. The composition of claim 12 , wherein said cytokine protein is selected from the group consisting of IL-2, IL-4, IL-12, IL-18, gamma-interferon, and a fusion protein thereof.
14. The composition of claim 12 , wherein said adjuvant is (a) IL-2, (b) a fusion protein containing IL-2 and the Fc portion of immunoglobulin G (IgG), or (c) a nucleic acid encoding (a) or (b).
15. The composition of claim 10 , wherein said construct further encodes another HIV protein or an immunogenic fragment thereof.
16. The composition of claim 15 , wherein said HIV protein is a surface protein of HIV viral particles.
17. The composition of claim 16 , wherein said HIV protein is an envelope protein.
18. An immunizing composition comprising:
a first nucleic acid encoding a mutant human immunodeficiency virus 1 (HIV1) sufficient for viral particle assembly but devoid of late domain motifs, wherein said mutant HIV1 comprises at least the membrane binding domain, major homology region, p2 domain, nucleocapsid domain, and the p1 domain of wild-type HIV1, and wherein expression of said first nucleic acid in a host cell results in the assembly of HIV1 virus particles that are incapable of budding from the surface of said host cell, and remain attached to the surface of said host cell; and
a second nucleic acid encoding a cytokine protein capable of stimulating a cytotoxic T lymphocyte (CTL) response.
19. The composition of claim 18 , wherein said first nucleic acid is capable of directing the expression of a mutant HIV Pr55Gag polypeptide in the absence of HIV Rev protein.
20. The composition of claim 19 , wherein said first nucleic acid has one or more mutations which decrease the effect of an inhibitory/instability sequence that is present in the corresponding nucleotide sequence of the native HIV GAG nucleic acid.
21. The composition of claim 18 , wherein said cytokine protein is selected from the group consisting of IL-2, IL-4, IL-12, IL-18, gamma-interferon, and a fusion protein thereof.
22. The composition of claim 18 , further comprising an adjuvant selected from IL-2 or a fusion protein containing IL-2 and the Fc portion of immunoglobulin G (IgG).
23. The composition of claim 18 , further comprising a third nucleic acid encoding another HIV protein.
24. An expression vector comprising the first and second nucleic acids of claim 18 .
25. The composition of claim 18 , wherein said first nucleic acid encodes a mutant HIV1 having mutations in the P(T/S)AP sequence motif (SEQ ID NO:1 or SEQ ID NO:2) of the GAGp6 domain that abolish binding to Tsg101.
26. The composition of claim 18 , wherein said first nucleic acid encodes a mutant HIV1 from which the P(T/S)AP sequence motif (SEQ ID NO:1 or SEQ ID NO:2) of the GAGp6 domain has been deleted.
27. The composition of claim 18 , wherein said first nucleic acid encodes a mutant HIV 1 devoid of late domain motifs.