Long lived T cells for treating HIV infection
A method of treating an HIV infected subject includes administering the subject an enriched CCR5 and/or CXCR4 gene edited CD4+ T cell population characterized by intermediate cells surface co-expression of CD45A and CD450 (RA int RO int ).
1 . A method of generating an enriched population of long-lived CD4 T cells that are devoid of a functional CCR5 and/or CXCR4 HIV co-receptor; the method comprising:
isolating CD4 T cells from a biological sample of a subject;
separating and/or enriching isolated CD4 T cells into cells having a CD45RA int CD45RO int phenotype and an increased expression of TCF7 compared to the isolated CD4 T cells prior to separation and/or enrichment, wherein enriching includes culturing the isolated CD4 T cells in the presence of IL-7 and IL-15 effective to promote expansion and/or formation of an enriched population of CD4 T cells having the CD45RA int CD45RO int phenotype, wherein the concentration of IL-7 and IL-15 is less than 10 ng/ml, and wherein at least 50% of the separated and/or enriched population of CD4 T cells have a CD45RA int CD45RO int CD95+CD127+CD27+ phenotype; and
modifying the CD4 T cells such that the CD4 T cells are devoid of a functional CCR5 and/or CXCR4 HIV co-receptor before separating and/or enriching the isolated CD4 T cells into cells having the CD45RA int CD45RO int phenotype and/or modifying the separated and/or enriched CD4 T cells having the CD45RA int CD45RO int phenotype such that the CD4 T cells having the CD45RA int CD45RO int phenotype are devoid of a functional CCR5 and/or CXCR4 HIV co-receptor.
2 . The method of claim 1 , wherein the biological sample comprises isolated peripheral blood mononuclear cells from the subject.
3 . The method of claim 1 , wherein the separated and/or enriched CD4 T cells express at least one of CD95, CD127, or CD27.
4 . The method of claim 1 , wherein the separated and/or enriched CD4 T cells intermediately express 4-1BB.
5 . The method of claim 1 , wherein the separated and/or enriched CD4 T cells express at least one of IL17RA, CD5, IL2RG, IGF2R, SLC38A1, IL7R, SLC44A2, SLC2A3, CD96, CD44, CD6, CCR4, IL4R, or SLC12A7.
6 . The method of claim 1 , wherein the separated and/or enriched CD4 T cells have a CD45RA int CD45RO int CD95+CD127+CD27+TCF7+ phenotype.
7 . The method of claim 1 , wherein the separated and/or enriched CD4 T cells have a
CD45RA int CD45RO int CD95+CD127+CD27+TCF7+CD44+ SCL38A1+IL2RG+CD6+CD5+ phenotype.
8 . The method of claim 1 , further comprising activating the isolated CD4 T cells with an anti-CD3 antibody and/or an anti-CD28 antibody.
9 . The method of claim 1 , further comprising culturing the enriched population of CD4 T cells having the CD45RA int CD45RO int phenotype in a culture medium comprising TGFβ and IL1β to maintain the CD45RA int CD45RO int phenotype.
10 . The method of claim 1 , wherein the CD4 T cells devoid of the functional CCR5 and/or CXCR4 HIV co-receptor are modified by inactivating a gene encoding CCR5 and/or CXCR4.
11 . The method of claim 10 , wherein the isolated T cells are genetically modified by at least one of transduction, transfection, and/or electroporation.
12 . An isolated enriched CD4+ T cell population, the T cell population characterized by intermediate cells surface co-expression of CD45A and CD450 (RA int RO int ) and expression of CD95+, CD127+, TCF7+ and CD27+, wherein the cells are modified such that they are devoid of a functional CCR5 and/or CXCR4 HIV co-receptor, wherein at least 50% of the enriched population of CD4+ T cells have a CD45RA int CD45RO int CD95+CD127+CD27+ phenotype.
13 . The population of claim 12 , upon administration to a subject infected with HIV being capable of promoting at least one of a sustained increase in absolute CD4 cell numbers, restoring HIV specific T cell immunity, or substantial decaying in HIV reservoir in the subject.
14 . A method of treating an HIV infected subject, the method comprising administering the subject the enriched T cell population of claim 12 .
15 . The method of claim 14 , where the subject has undergone and/or continues to undergo antiretroviral therapy.
16 . The method of claim 14 , wherein the enriched T cell population is administered to the subject at an amount effective to promote at least one of a sustained increase in absolute CD4 cell numbers, restoration of HIV specific T cell immunity, or a substantial decay in HIV reservoir in the subject.