Stem cell screening and transplantation therapy for hiv infection
This invention provides methods for preventing or treating any disease arising from HIV infection, including AIDS and AIDS-related complex (ARC). The method comprises screening a plurality of donors to identify stem cells with a beneficial gene or genes and then transplanting the therapeutic stem cells into a patient. In preferred embodiments, the beneficial gene encodes a polymorphism that renders cells refractory to HIV infection. The polymorphism may be for a gene that encodes a ligand of a receptor for HIV entry, a product of the HLA complex, or a receptor for HIV entry. The Invention also provides isolated populations of stem cells with beneficial genes for resisting infections.
1 . A method for preventing or treating HIV infection, said method comprising:
a) screening a plurality of cells to identify stem cells having a beneficial gene; and
b) transplanting said stem cells into a patient, thereby preventing or treating said HIV infection.
2 . The method of claim 1 , wherein said beneficial gene is a polymorphism of a gene encoding a protein expressed by immune cells.
3 . The method of claim 1 , wherein said beneficial gene is one which reduces the ability of HIV to infect an immune cell.
4 . The method of claim 1 , wherein said beneficial gene is one which enhances the ability of an immune cell to neutralize the virus through immune reconstitution.
5 . The method of claim 2 , wherein said protein is a ligand of a receptor for HIV entry.
6 . The method of claim 5 , wherein said ligand is SDF-1 alpha and said polymorphism is SDF-1 alpha 3′A.
7 . The method of claim 5 , wherein said ligand is RANTES and said polymorphism is in the promoter region and increases expression levels.
8 . The method of claim 2 , wherein said protein is encoded by a gene in the HLA complex.
9 . The method of claim 8 , wherein said protein encoded by a gene in the HLA complex is selected from the group consisting of MHC class I molecule, MHC class II molecule, TNF, and complement.
10 . The method of claim 2 , wherein said protein is a receptor or coreceptor for HIV entry.
11 . The method of claim 10 , wherein said receptor for HIV entry is CD4.
12 . The method of claim 10 , wherein said coreceptor for HIV entry is CCR2.
13 . The method of claim 12 , wherein said polymorphism is CCR2-64I.
14 . The method of claim 10 , wherein said coreceptor for HIV entry is CCR5.
15 . The method of claim 14 , wherein said polymorphism is a 32 basepair deletion in the coding region.
16 . The method of claim 14 , wherein said polymorphism is CCR5 m303.
17 . The method of claim 14 , wherein said polymorphism is in the promoter region of CCR5.
18 . The method of claim 14 , wherein the HIV infection is caused by predominantly monocytotropic HIV isolates.
19 . The method of claim 1 , wherein said plurality of cells are obtained from the group consisting of embryos, marrow, peripheral blood, placental blood, umbilical cord blood, and adipose tissue.
20 . The method of claim 1 , further comprising in vitro or in vivo expansion of said stem cells.
21 . The method of claim 1 , wherein said method further comprises identification of the HLA genotype or phenotype of said stem cells.
22 . The method of claim 21 , wherein said identification of the HLA genotype is via a high-throughput method using allele-specific primers and HLA locus-specific capture oligonucleotides immobilized on a solid phase.
23 . The method of claim 1 , wherein said screening comprises identification of stem cells expressing the protein product of said beneficial gene.
24 . The method of claim 23 , wherein said protein product is detected or identified using an immunological assay.
25 . The method of claim 1 , wherein said screening comprises identification of stem cells with said beneficial gene.
26 . The method of claim 25 , wherein said beneficial gene is detected using a hybridization-based assay, a sequencing assay, or a functional assay.
27 . The method of claim 1 , further comprising treatment of said stem cells to express a non-native HLA protein or to inhibit expression of the native HLA protein.
28 . An isolated population of human cells comprising
viable human hematopoietic stem cells,
wherein the stem cells comprise a beneficial gene that confers resistance to HIV infection.
29 . The isolated population of human cells of claim 28 , wherein the stem cells are derived from a member of the group consisting of embryos, bone marrow, peripheral blood, placental blood, umbilical cord blood, and adipose tissue
30 . The isolated population of human cells of claim 28 , wherein the beneficial gene is a polymorphism of CCR5.
31 . The isolated population of human cells of claim 30 , wherein the beneficial gene is a polymorphism of CCR5 selected from the group consisting of CCR5 delta 32, CCR5 m303, a polymorphism in the CCR5 promoter, and CCR5R60S.
32 . The isolated population of human cells of claim 31 , wherein the beneficial gene is homozygous for the CCR5 delta 32 polymorphism.
33 . The isolated population of human cells of claim 31 , wherein the beneficial gene is heterozygous for the CCR5 delta 32 polymorphism.
34 . The isolated population of human cells of claim 28 , wherein the stem cells further comprise an HLA genotype that compatible with a recipient of the stem cells.
35 . The isolated population of human cells of claim 28 , further comprising a pharmaceutical carrier.