# US 2021/0330783 A1 · Hiv pre-immunization and immunotherapy > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0330783 A1 - Document ID: US20210330783A1 - Status: Published application - Kind: A1 - Application number: 17/175,278 - Filed: 2021-02-12 - Published: 2021-10-28 - Earliest claimed: 2016-07-08 - Assignees: American Gene Technologies International Inc. - Applicants: American Gene Technologies International Inc. - Inventors: Charles David Pauza; Haishan Li; Tyler Lahusen; Jeff Galvin - Web page: https://patentgrep.com/patent/US20210330783A1 ## CPC - A61K 39/21: Retroviridae, e.g. equine infectious anemia virus - A61K 31/7105: Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links - A61K 35/14: Blood; Artificial blood (perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42) - A61K 39/12: Viral antigens - A61K 39/39: characterised by the immunostimulating additives, e.g. chemical adjuvants - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/46: Viral antigens - A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca - A61P 31/18: for HIV - C07K 14/7158: {for chemokines} - C12N 5/0634: {Cells from the blood or the immune system} - C12N 5/0636: {T lymphocytes} - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids} - C12N 15/1132: {against retroviridae, e.g. HIV} - C12N 15/1138: {against receptors or cell surface proteins} - C12N 15/86: Viral vectors - A61K 2035/124: {the cells being hematopoietic, bone marrow derived or blood cells} - A61K 2039/545: {characterised by the dose, timing or administration schedule} - A61K 2039/55561: {CpG containing adjuvants; Oligonucleotide containing adjuvants} - A61K 2239/38: characterised by the dose, timing or administration schedule - C12N 2310/122: Hairpin - C12N 2310/14: interfering nucleic acids [NA] - C12N 2310/141: MicroRNAs, miRNAs - C12N 2320/32: Special delivery means, e.g. tissue-specific - C12N 2330/51: Specially adapted vectors - C12N 2510/00: Genetically modified cells - C12N 2710/24143: viral genome or elements thereof as genetic vector - C12N 2740/15021: Viruses as such, e.g. new isolates, mutants or their genomic sequences - C12N 2740/15034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2740/15043: viral genome or elements thereof as genetic vector - C12N 2740/15052: relating to complementing cells and packaging systems for producing virus or viral particles - C12N 2740/16022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2740/16034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2740/16043: viral genome or elements thereof as genetic vector - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract 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. ## Claims ### Claim 1 A lentiviral vector comprising at least one encoded genetic element, wherein the at least one encoded genetic element comprises each of: (i) a sequence comprising at least 80% sequence identity with SEQ ID NO: 2 and (ii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 108; or two of: (iii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 97, (iv) a sequence comprising at least 80% sequence identity with SEQ ID NO: 6, and (v) a sequence comprising at least 80% sequence identity with SEQ ID NO: 7. ### Claim 2 (depends on 2) The lentiviral vector of claim 2, wherein the at least one encoded genetic element comprises: each of: SEQ ID NO: 2 and SEQ ID NO: 108 or two of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7. ### Claim 3 (depends on 1) A lentiviral vector system for expressing a lentiviral particle, the system comprising: a lentiviral vector according to claim 1; an envelope plasmid for expressing an envelope protein optimized for infecting a cell; and at least one helper plasmid for expressing gag, pol, and rev genes, wherein the lentiviral vector, the envelope plasmid, and the at least one helper plasmid are capable of transfecting a packaging cell line, and wherein, upon such transfection, the packaging cell line is capable of producing a lentiviral particle capable of inhibiting production of chemokine receptor CCR5 or targeting an HIV RNA sequence. ### Claim 4 (depends on 3) The lentiviral vector system of claim 3, wherein the at least one helper plasmid comprises a first plasmid for expressing the gag and pol genes, and a second plasmid for expressing the rev gene. ### Claim 5 (depends on 1) A lentiviral particle capable of infecting a cell, the lentiviral particle comprising an envelope protein optimized for infecting a cell, and a lentiviral vector according to claim 1. ### Claim 6 (depends on 5) The lentiviral particle of claim 5, wherein the envelope protein is optimized for infecting a T cell. ### Claim 7 (depends on 6) The lentiviral particle of claim 6, wherein the envelope protein is optimized for infecting a CD4+ T cell. ### Claim 8 (depends on 5) A modified cell comprising a CD4+ T cell, wherein the CD4+ T cell has been transfected with a lentiviral particle according to claim 5. ### Claim 9 (depends on 8) The modified cell of claim 8, wherein the CD4+ T cell is capable of recognizing an HIV antigen. ### Claim 10 (depends on 9) The modified cell of claim 9, wherein the HIV antigen comprises a gag antigen. ### Claim 11 (depends on 8) The modified cell of claim 8, wherein the CD4+ T cell expresses a decreased level of CCR5 following infection with the lentiviral particle. ### Claim 12 A method of treating cells infected with HIV, the method comprising: (a) obtaining, or having obtained, peripheral blood mononuclear cells (PBMC) from a subject infected with HIV, wherein the subject has been previously immunized with a therapeutically effective amount of a first stimulatory agent; (b) contacting, or having contacted, the PBMC with a therapeutically effective amount of a second stimulatory agent, wherein the contacting is carried out ex vivo; (c) transducing, or having transduced, the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein at least one encoded genetic element comprises: each of: (i) a sequence comprising at least 80% sequence identity with SEQ ID NO: 2 and (ii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 108; or two of: (iii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 97, (iv) a sequence comprising at least 80% sequence identity with SEQ ID NO: 6, and (v) a sequence comprising at least 80% sequence identity with SEQ ID NO: 7; and (d) culturing, or having cultured, the transduced PBMC for at least 1 day. ### Claim 13 (depends on 12) The method of claim 12, wherein the transduced PBMC are cultured from about 1 to about 35 days. ### Claim 14 (depends on 12) The method of claim 12, further comprising infusing, or having infused, the transduced PBMC into a subject. ### Claim 15 (depends on 14) The method of claim 14, wherein the subject is a human. ### Claim 16 (depends on 12) The method of claim 12, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a peptide. ### Claim 17 (depends on 16) The method of claim 16, wherein the peptide comprises a gag peptide. ### Claim 18 (depends on 12) The method of claim 12, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a vaccine. ### Claim 19 (depends on 18) The method of claim 18, wherein the vaccine comprises an HIV vaccine. ### Claim 20 (depends on 19) The method of claim 19, wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof. ### Claim 21 (depends on 12) The method of claim 12, wherein the first stimulatory agent and the second stimulatory agent are the same. ### Claim 22 (depends on 12) The method of claim 12, wherein the viral delivery system comprises a lentiviral particle. ### Claim 23 (depends on 12) The method of claim 12, wherein the at least one genetic element, when expressed, is capable of targeting an HIV RNA sequence. ### Claim 24 (depends on 23) The method of claim 23, wherein, when the at least one encoded genetic element comprises a sequence having at least 80% sequence identity with SEQ ID NO: 97, the at least one genetic element, when expressed, is also capable of inhibiting production of chemokine receptor CCR5. ### Claim 25 (depends on 12) The method of claim 12, wherein the at least one encoded genetic element comprises: each of: SEQ ID NO: 2 and SEQ ID NO: 108 or two of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7. ### Claim 26 A method of treating HIV infection in a subject, the method comprising: (a) removing, or having removed, leukocytes from the subject, wherein the subject has been previously immunized with a therapeutically effective amount of a first stimulatory agent; (b) purifying, or having purified, peripheral blood mononuclear cells (PBMC) ex vivo from the leukocytes; (c) contacting, or having contacted, the PBMC ex vivo with a therapeutically effective amount of a second stimulatory agent; (d) transducing, of having transduced, the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein the at least one genetic element comprises: each of: (i) a sequence comprising at least 80% sequence identity with SEQ ID NO: 2 and (ii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 108; or two of: (iii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 97, (iv) a sequence comprising at least 80% sequence identity with SEQ ID NO: 6, and (v) a sequence comprising at least 80% sequence identity with SEQ ID NO: 7; and (e) culturing, or having cultured, the transduced PBMC for at least 1 day. ### Claim 27 (depends on 26) The method of claim 26, wherein the transduced PBMC are cultured from about 1 to about 35 days. ### Claim 28 (depends on 26) The method of claim 26, further comprising infusing, or having infused, the transduced PBMC into the subject. ### Claim 29 (depends on 28) The method of claim 28, wherein the subject is a human. ### Claim 30 (depends on 26) The method of claim 26, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a peptide. ### Claim 31 (depends on 30) The method of claim 30, wherein the peptide comprises a gag peptide. ### Claim 32 (depends on 26) The method of claim 26, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a vaccine. ### Claim 33 (depends on 32) The method of claim 32, wherein the vaccine comprises an HIV vaccine. ### Claim 34 (depends on 33) The method of claim 33, wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof. ### Claim 35 (depends on 26) The method of claim 26, wherein the first stimulatory agent and the second stimulatory agent are the same. ### Claim 36 (depends on 26) The method of claim 26, wherein the viral delivery system comprises a lentiviral particle. ### Claim 37 (depends on 26) The method of claim 26, wherein the at least one genetic element comprises at least one small RNA capable of targeting an HIV RNA sequence. ### Claim 38 (depends on 26) The method of claim 26, wherein the at least one genetic element comprises: each of: SEQ ID NO: 2, and SEQ ID NO: 108; or two of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7. ### Claim 39 A lentiviral vector comprising an encoded microRNA cluster, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 90% sequence identity with SEQ ID NO: 1, (ii) at least 90% sequence identity with SEQ ID NO: 2, and (iii) at least 90% sequence identity with SEQ ID NO: 108. ### Claim 40 (depends on 39) The lentiviral vector of claim 39, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 95% sequence identity with SEQ ID NO: 1, (ii) at least 95% sequence identity with SEQ ID NO: 2, or (iii) at least 95% sequence identity with SEQ ID NO: 108. ### Claim 41 (depends on 39) The lentiviral vector of claim 39, wherein the encoded microRNA cluster comprises a sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 108. ### Claim 42 A lentiviral particle capable of infecting a target cell, the lentiviral particle comprising: (a) an envelope protein capable of infecting the target cell; and (b) an encoded microRNA cluster, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 90% sequence identity with SEQ ID NO: 1, (ii) at least 90% sequence identity with SEQ ID NO: 2, and (iii) at least 90% sequence identity with SEQ ID NO: 108. ### Claim 43 (depends on 42) The lentiviral particle of claim 42, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 95% sequence identity with SEQ ID NO: 1, (ii) at least 95% sequence identity with SEQ ID NO: 2, or (iii) at least 95% sequence identity with SEQ ID NO: 108. ### Claim 44 (depends on 42) The lentiviral particle of claim 42, wherein the encoded microRNA cluster comprises a sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 108. ### Claim 45 (depends on 42) The lentiviral particle of claim 42, wherein the target cell is a CD4+ T cell. ### Claim 46 A modified cell comprising a primary T cell infected with a lentiviral particle, wherein the lentiviral particle comprises: (a) an envelope protein capable of infecting the target cell; and (b) an encoded microRNA cluster, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 90% sequence identity with SEQ ID NO: 1, (ii) at least 90% sequence identity with SEQ ID NO: 2, and (iii) at least 90% sequence identity with SEQ ID NO: 108. ### Claim 47 (depends on 46) The modified cell of claim 46, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 95% sequence identity with SEQ ID NO: 1, (ii) at least 95% sequence identity with SEQ ID NO: 2, or (iii) at least 95% sequence identity with SEQ ID NO: 108. ### Claim 48 (depends on 46) The modified cell of claim 46, wherein the encoded microRNA cluster comprises a sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 108. ### Claim 49 A method of treating cells infected with HIV, the method comprising: (a) contacting or having contacted peripheral blood mononuclear cells (PBMC) isolated from a subject infected with HIV with a therapeutically effective amount of an ex vivo stimulatory agent, wherein the contacting is conducted ex vivo; (b) transducing or having transduced the PBMC ex vivo with a lentiviral particle, wherein the lentiviral particle comprises: (i) an envelope protein capable of infecting the PBMC; and (ii) an encoded microRNA cluster, wherein the encoded microRNA cluster comprises a sequence comprising (a) at least 90% sequence identity with SEQ ID NO: 1, (b) at least 90% sequence identity with SEQ ID NO: 2, and (c) at least 90% sequence identity with SEQ ID NO: 108. ### Claim 50 (depends on 49) The method of claim 49, wherein the encoded microRNA cluster comprises a sequence comprising (i) at least 95% sequence identity with SEQ ID NO: 1, (ii) at least 95% sequence identity with SEQ ID NO: 2, or (iii) at least 95% sequence identity with SEQ ID NO: 108. ### Claim 51 (depends on 49) The method of claim 49, wherein the encoded microRNA cluster comprises a sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 108. ### Claim 52 (depends on 49) The method of claim 49, further comprising infusing or having infused the transduced PBMC into a subject. ### Claim 53 (depends on 49) The method of claim 49, further comprising positively selecting or having positively selected HIV-specific CD4+ T cells from the PBMC. ### Claim 54 (depends on 49) The method of claim 49, further comprising immunizing or having immunized the subject with an effective amount of an in vivo stimulatory agent, wherein the immunization occurs prior to contacting the peripheral blood mononuclear cells (PBMC) with the ex vivo stimulatory agent. ### Claim 55 (depends on 54) The method of claim 54, wherein each of the in vivo stimulatory agent and ex vivo stimulatory agent is independently selected from a peptide and a vaccine. ### Claim 56 A lentiviral vector comprising at least one encoded genetic element, wherein the at least one encoded genetic element comprises at least two of: (i) a sequence comprising at least 80% sequence identity with SEQ ID NO: 1, (ii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 2, and (iii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 108; or each of: (iv) a sequence comprising at least 80% sequence identity with SEQ ID NO: 97, (v) a sequence comprising at least 80% sequence identity with SEQ ID NO: 6, and (vi) a sequence comprising at least 80% sequence identity with SEQ ID NO: 7. ### Claim 57 (depends on 56) The lentiviral vector of claim 56, wherein the at least one encoded genetic element comprises: at least two of: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 108; or each of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7. ### Claim 58 (depends on 56) A lentiviral vector system for expressing a lentiviral particle, the system comprising: a lentiviral vector according to claim 56; an envelope plasmid for expressing an envelope protein optimized for infecting a cell; and at least one helper plasmid for expressing gag, pol, and rev genes, wherein the lentiviral vector, the envelope plasmid, and the at least one helper plasmid are capable of transfecting a packaging cell line, and wherein, upon such transfection, the packaging cell line is capable of producing a lentiviral particle capable of inhibiting production of chemokine receptor CCR5 or targeting an HIV RNA sequence. ### Claim 59 (depends on 58) The lentiviral vector system of claim 58, wherein the at least one helper plasmid comprises a first plasmid for expressing the gag and pol genes, and a second plasmid for expressing the rev gene. ### Claim 60 (depends on 56) A lentiviral particle capable of infecting a cell, the lentiviral particle comprising an envelope protein optimized for infecting a cell, and a lentiviral vector according to claim 56. ### Claim 61 (depends on 60) The lentiviral particle of claim 60, wherein the envelope protein is optimized for infecting a T cell. ### Claim 62 (depends on 61) The lentiviral particle of claim 61, wherein the envelope protein is optimized for infecting a CD4+ T cell. ### Claim 63 (depends on 60) A modified cell comprising a CD4+ T cell, wherein the CD4+ T cell has been transfected with a lentiviral particle according to claim 60. ### Claim 64 (depends on 63) The modified cell of claim 63, wherein the CD4+ T cell is capable of recognizing an HIV antigen. ### Claim 65 (depends on 64) The modified cell of claim 64, wherein the HIV antigen comprises a gag antigen. ### Claim 66 (depends on 63) The modified cell of claim 63, wherein the CD4+ T cell expresses a decreased level of CCR5 following infection with the lentiviral particle. ### Claim 67 A method of treating cells infected with HIV, the method comprising: (a) obtaining, or having obtained, peripheral blood mononuclear cells (PBMC) from a subject infected with HIV, wherein the subject has been previously immunized with a therapeutically effective amount of a first stimulatory agent; (b) contacting, or having contacted, the PBMC with a therapeutically effective amount of a second stimulatory agent, wherein the contacting is carried out ex vivo; (c) transducing, or having transduced, the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein at least one encoded genetic element comprises: at least two of: (i) a sequence comprising at least 80% sequence identity with SEQ ID NO: 1, (ii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 2, and (iii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 108; or each of: (iv) a sequence comprising at least 80% sequence identity with SEQ ID NO: 97, (v) a sequence comprising at least 80% sequence identity with SEQ ID NO: 6, and (vi) a sequence comprising at least 80% sequence identity with SEQ ID NO: 7; and (d) culturing, or having cultured, the transduced PBMC for at least 1 day. ### Claim 68 (depends on 67) The method of claim 67, wherein the transduced PBMC are cultured from about 1 to about 35 days. ### Claim 69 (depends on 67) The method of claim 67, further comprising infusing, or having infused, the transduced PBMC into a subject. ### Claim 70 (depends on 69) The method of claim 69, wherein the subject is a human. ### Claim 71 (depends on 67) The method of claim 67, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a peptide. ### Claim 72 (depends on 71) The method of claim 71, wherein the peptide comprises a gag peptide. ### Claim 73 (depends on 67) The method of claim 67, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a vaccine. ### Claim 74 (depends on 73) The method of claim 73, wherein the vaccine comprises an HIV vaccine. ### Claim 75 (depends on 74) The method of claim 74, wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof. ### Claim 76 (depends on 67) The method of claim 67, wherein the first stimulatory agent and the second stimulatory agent are the same. ### Claim 77 (depends on 67) The method of claim 67, wherein the viral delivery system comprises a lentiviral particle. ### Claim 78 (depends on 67) The method of claim 67, wherein the at least one genetic element, when expressed, is capable of targeting an HIV RNA sequence. ### Claim 79 (depends on 78) The method of claim 78, wherein, when the at least one encoded genetic element comprises a sequence having at least 80% sequence identity with SEQ ID NO: 1, or a sequence having at least 80% sequence identity with SEQ ID NO: 97, the at least one genetic element, when expressed, is also capable of inhibiting production of chemokine receptor CCR5. ### Claim 80 (depends on 67) The method of claim 67, wherein the at least one encoded genetic element comprises: at least two of: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 108; or each of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7. ### Claim 81 A method of treating HIV infection in a subject, the method comprising: (a) removing, or having removed, leukocytes from the subject, wherein the subject has been previously immunized with a therapeutically effective amount of a first stimulatory agent; (b) purifying, or having purified, peripheral blood mononuclear cells (PBMC) ex vivo from the leukocytes; (c) contacting, or having contacted, the PBMC ex vivo with a therapeutically effective amount of a second stimulatory agent; (d) transducing, of having transduced, the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein the at least one genetic element comprises: at least two of: (i) a sequence comprising at least 80% sequence identity with SEQ ID NO: 1, (ii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 2, and (iii) a sequence comprising at least 80% sequence identity with SEQ ID NO: 108; or each of: (iv) a sequence comprising at least 80% sequence identity with SEQ ID NO: 97, (v) a sequence comprising at least 80% sequence identity with SEQ ID NO: 6, and (vi) a sequence comprising at least 80% sequence identity with SEQ ID NO: 7; and (e) culturing, or having cultured, the transduced PBMC for at least 1 day. ### Claim 82 (depends on 81) The method of claim 81, wherein the transduced PBMC are cultured from about 1 to about 35 days. ### Claim 83 (depends on 81) The method of claim 81, further comprising infusing, or having infused, the transduced PBMC into the subject. ### Claim 84 (depends on 81) The method of claim 81, wherein the subject is a human. ### Claim 85 (depends on 81) The method of claim 81, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a peptide. ### Claim 86 (depends on 85) The method of claim 85, wherein the peptide comprises a gag peptide. ### Claim 87 (depends on 81) The method of claim 81, wherein at least one of the first stimulatory agent and the second stimulatory agent comprises a vaccine. ### Claim 88 (depends on 87) The method of claim 87, wherein the vaccine comprises an HIV vaccine. ### Claim 89 (depends on 88) The method of claim 88, wherein the HIV vaccine comprises a MVA/HIV62B vaccine or a variant thereof. ### Claim 90 (depends on 81) The method of claim 81, wherein the first stimulatory agent and the second stimulatory agent are the same. ### Claim 91 (depends on 81) The method of claim 81, wherein the viral delivery system comprises a lentiviral particle. ### Claim 92 (depends on 81) The method of claim 81, wherein the at least one genetic element comprises at least one small RNA capable of targeting an HIV RNA sequence. ### Claim 93 (depends on 92) The method of claim 92, wherein, when the at least one genetic element comprises a sequence having at least 80% sequence identity with SEQ ID NO: 1, or a sequence having at least 80% sequence identity with SEQ ID NO: 97, the at least one genetic element, when expressed, is also capable of inhibiting production of chemokine receptor CCR5. ### Claim 94 (depends on 81) The method of claim 81, wherein the at least one genetic element comprises: at least two of: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 108; or each of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7.