HIV pre-immunization and immunotherapy
The present invention relates generally to immunization and immunotherapy for the treatment or prevention of HIV. In particular, the methods include in vivo and/or ex vivo enrichment of HIV-specific CD4+ T cells.
1. A method of treating cells, the method comprising:
(a) positively selecting HIV-specific CD4+ T cells from PBMC isolated from a subject infected with HIV;
(b) contacting the CD4+ T cells with a therapeutically effective amount of a stimulatory agent, wherein the contacting is carried out ex vivo,
wherein the stimulatory agent comprises a peptide, a mixture of peptides, or a vaccine;
(c) transducing the CD4+ T cells ex vivo with a viral delivery system encoding at least one genetic element, wherein the at least one genetic element comprises (i) a sequence having at least 90% sequence identity with SEQ ID NO: 6, (ii) a sequence having at least 90% sequence identity with SEQ ID NO: 7, and (iii) a sequence having at least 90% sequence identity with SEQ ID NO: 97, wherein the at least one genetic element does not comprise any small RNA targeting HIV sequences other than Vif and Tat; and
(d) culturing the transduced CD4+ T cells for at least 1 day.
2. The method of claim 1 , further comprising infusing the transduced CD4+ T cells into a subject.
3. The method of claim 1 , wherein the stimulatory agent comprises a peptide.
4. The method of claim 3 , wherein the peptide comprises a gag peptide.
5. The method of claim 1 , wherein the stimulatory agent comprises a vaccine.
6. The method of claim 5 , wherein the vaccine comprises a HIV vaccine.
7. The method of claim 6 , wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof.
8. The method of claim 1 , wherein the at least one genetic element comprises at least one microRNA.
9. The method of claim 8 , wherein the at least one microRNA comprises a microRNA cluster.
10. The method of claim 1 , wherein the at least one genetic element comprises SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 97.
11. A method of treating HIV infection in a subject, the method comprising:
(a) immunizing the subject with an effective amount of a first stimulatory agent;
(b) purifying peripheral blood mononuclear cells (PBMC) isolated from the subject;
(c) positively selecting HIV-specific CD4+ T cells from the PBMC;
(d) contacting the HIV-specific CD4+ T-cells ex vivo with a therapeutically effective amount of a second stimulatory agent,
wherein the second stimulatory agent comprises a peptide, a mixture of peptides, or a vaccine;
(e) transducing the HIV-specific CD4+ T-cells ex vivo with a viral delivery system encoding at least one genetic element,
wherein the at least one genetic element comprises (i) a sequence having at least 90% sequence identity with SEQ ID NO: 6, (ii) a sequence having at least 90% sequence identity with SEQ ID NO: 7, and (iii) a sequence having at least 90% sequence identity with SEQ ID NO: 97, wherein the at least one genetic element does not comprise any small RNA targeting HIV sequences other than Vif and Tat; and
(f) culturing the transduced HIV-specific CD4+ T-cells for at least 1 day; and
(q) infusing the transduced HIV-specific CD4+ T-cells into the subject.
12. The method of claim 11 , wherein the first and second stimulatory agents are the same.
13. The method of claim 11 , wherein at least one of the first and second stimulatory agents comprises a HIV vaccine.
14. The method of claim 13 , wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof.
15. The method of claim 11 , wherein the viral delivery system comprises a lentiviral particle.
16. The method of claim 11 , wherein the at least one genetic element comprises SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 97.
17. A method of treating HIV infection in a subject, the method comprising:
(a) immunizing the subject with an effective amount of an HIV vaccine;
(b) purifying peripheral blood mononuclear cells (PBMC) isolated from the subject;
(c) positively selecting HIV-specific CD4+ T cells from the PBMC;
(d) contacting the HIV-specific CD4+ T cells ex vivo with a therapeutically effective amount of a second stimulatory agent,
wherein the second stimulatory agent comprises a peptide, a mixture of peptides, or a vaccine;
(e) transducing the HIV-specific CD4+ T cells ex vivo with a viral delivery system encoding at least one genetic element,
wherein the at least one genetic element comprises (i) a sequence having at least 90% sequence identity with SEQ ID NO: 6, (ii) a sequence having at least 90% sequence identity with SEQ ID NO: 7, and (iii) a sequence having at least 90% sequence identity with SEQ ID NO: 97, wherein the at least one genetic element does not comprise any small RNA targeting HIV sequences other than Vif and Tat; and
(f) culturing the transduced HIV-specific CD4+ T cells for at least 1 day; and
(q) infusing the transduced HIV-specific CD4+ T-cells into the subject.
18. The method of claim 17 , wherein the at least one genetic element comprises SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 97.