Human immunodeficiency virus infection of human placental cord blood CD34+AC133+ stem cells and their progeny.
The AC133 is a novel antigen selectively expressed on primitive CD34bright stem cells and is a valuable marker for the selection of long-term culture-initiating cells (LTC-ICs) and severe combined immunodeficiency (SCID)-repopulating cells. Human placental cord blood (HPCB) is a rich source of CD34+AC133+ cells. Since AC133 antibody is likely to be used as an alternative to CD34 for the selection of stem cells in transplant and gene therapy situations, we examined the susceptibility of HPCB-isolated CD34+AC133+ stem cells to infection with free and cell-associated HIV-1 in vitro. Freshly isolated HPCB CD34+AC133+ stem cells were not susceptible to HIV-1 infection as determined by PCR and reverse transcriptase assays. Inoculation with HIV-1 did not affect the viability and clonogenic ability of HPCB CD34+AC133+ cells. Although the highly purified HPCB CD34+AC133+ stem cells contained mRNA for CD4 and CXCR4 receptors, CD4 and CXCR4 proteins were not expressed on these cells. Similarly, CCR5 protein, the major macrophage-tropic HIV-1 coreceptor, was not expressed in freshly isolated HPCB CD34+AC133+ stem cells, although the transcript for CCR5 was identified in these cells. Expression of CD4, CXCR4, and CCR5 receptor proteins on the progeny derived from HPCB CD34+AC133+ stem cells was detected and correlated with susceptibility to HIV-1 infection in vitro. These findings suggest that freshly isolated HPCB CD34+AC133+ stem cells are not susceptible to HIV-1 infection and may not be a viral reservoir. These data have important implications for the use of AC133 antibody as a means of enriching for primitive hematopoietic stem cells from placental cord blood and in the design of stem cell or progenitor cell-based gene therapeutic strategies for perinatal HIV-1 infection.
Hariharan
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D
Department of Pediatrics, University of Pennsylvania Medical School, Philadelphia 19104, USA.
Li
Y
Y
Campbell
D E
DE
Douglas
S D
SD
Starr
S E
SE
Ho
W
W
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000378:metabolism*; D015704:CD4 Antigens / Q000235:genetics / Q000378:metabolism; D003114:Colony-Forming Units Assay; D004279:DNA, Viral / Q000032:analysis; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism*; D054303:HIV Reverse Transcriptase / Q000032:analysis; D015497:HIV-1 / Q000302:isolation & purification; D006801:Humans; D010455:Peptides / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000032:analysis; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000276:immunology* / Q000821:virology*; D013997:Time Factors
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D015704:CD4 Antigens; D004279:DNA, Viral; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D012333:RNA, Messenger; D019713:Receptors, CCR5; D019718:Receptors, CXCR4; D054303:HIV Reverse Transcriptase