Methods of producing cells resistant to HIV infection
The present invention relates generally to immunotherapy for preventing HIV infection in HIV-negative individuals. In particular, the methods include methods for making cells resistant to HIV and administering an immune therapy to an HIV-negative subject to prevent an HIV infection.
1. A method of producing cells that are resistant to HIV infection, the method comprising:
(a) contacting peripheral blood mononuclear cells (PBMC) isolated from a subject that is HIV-negative with a therapeutically effective amount of a stimulatory agent, wherein the contacting is carried out ex vivo;
(b) transducing the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein the at least one genetic element comprises a sequence encoding:
a microRNA capable of inhibiting production of chemokine receptor CCR5 comprising a sequence having at least 80% identity with SEQ ID NO: 97;
a microRNA capable of inhibiting expression of HIV Vif comprising a sequence having at least 80% identity with SEQ ID NO: 6; and
a microRNA capable of inhibiting HIV Tat comprising a sequence having at least 80% identity with SEQ ID NO: 7, wherein the genetic element does not encode for a microRNA capable of inhibiting an HIV gene other than Vif and Tat, and
(c) culturing the transduced PBMC for at least 1 day.
2. The method of claim 1 , further comprising infusing the transduced PBMC into a subject.
3. The method of claim 1 , wherein the stimulatory agent comprises a gag peptide.
4. The method of claim 1 , wherein the stimulatory agent comprises an HIV vaccine.
5. The method of claim 4 , wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof.
6. The method of claim 1 , wherein the viral delivery system comprises a lentiviral particle.
7. The method of claim 1 , wherein the HIV RNA sequence comprises an HIV Vif sequence, an HIV Tat sequence, or a variant thereof.
8. The method of claim 1 , wherein the genetic element comprises each of: (i) a sequence comprising SEQ ID NO: 97; (ii) a sequence comprising SEQ ID NO: 6; and (iii) a sequence comprising SEQ ID NO: 7.
9. A method of inhibiting HIV infection in a HIV-negative subject, the method comprising:
(a) immunizing the subject with an effective amount of a first stimulatory agent;
(b) removing leukocytes from the subject and purifying peripheral blood mononuclear cells (PBMC);
(c) contacting the PBMC ex vivo with a therapeutically effective amount of a second stimulatory agent;
(d) transducing the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein the at least one genetic element comprises a sequence encoding:
a microRNA capable of inhibiting production of chemokine receptor CCR5 comprising a sequence having at least 80% identity with SEQ ID NO: 97;
a microRNA capable of inhibiting expression of HIV Vif comprising a sequence having at least 80% identity with SEQ ID NO: 6; and
a microRNA capable of inhibiting HIV Tat comprising a sequence having at least 80% identity with SEQ ID NO: 7, wherein the genetic element does not encode for a microRNA capable of inhibiting an HIV gene other than Vif and Tat;
(e) culturing the transduced PBMC for at least 1 day; and
(f) infusing the transduced PBMC into the subject.
10. The method of claim 9 , wherein at least one of the first and second stimulatory agents comprises a gag peptide.
11. The method of claim 9 , wherein at least one of the first and second stimulatory agents comprises an HIV vaccine.
12. The method of claim 11 , wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof.