Modified antisense oligodeoxynucleotides against the splice acceptor site of tat do not inhibit in vitro hematopoietic colony growth in HIV-positive patients.
The hematopoietic failure in the majority of patients with progressive HIV infection is further aggravated by virustatic agents like azidothymidine. As an alternative therapeutic attempt, three derivatives of an antisense oligodeoxynucleotide (ODN) against the splice acceptor site of the tat gene have been shown to inhibit HIV replication in vitro. This study was aimed at examining whether these agents are toxic to the hematopoietic progenitor cells. To this end, bone marrow cells from HIV-positive and healthy persons were depleted from adherent cells to eliminate fibroblasts. In further experiments, the cells were additionally enriched for CD34-positive hematopoietic progenitor cells or were depleted from delta TCS-1-positive T lymphocytes. At concentrations of 1.25-10 microM, the three antisense ODN did not inhibit any erythrocyte or granulocyte-monocyte colony growth from CD34-positive cells, either from the HIV-positive or from the HIV-negative cohort. In contrast to azidothymidine, which served as inhibitory control, a significant increase of colony growth was seen after depletion of fibroblasts, of delta TCS-1-positive cells, or without cell separation. In conclusion, the three oligodeoxynucleotides do not exert any hematotoxic effect but do increase colony formation from low-density bone marrow cells in vitro and could therefore be useful in future clinical studies.
Geissler
R G
RG
Department of Hematology, University of Frankfurt/Main, Germany.
Muth
J
J
Maurer
A
A
Mentzel
U
U
Mag
M
M
Engels
J W
JW
Hoelzer
D
D
Ganser
A
A
D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34; D001483:Base Sequence; D001853:Bone Marrow / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D015672:Erythroid Precursor Cells / Q000473:pathology; D015756:Genes, tat; D006098:Granulocytes / Q000473:pathology; D006679:HIV Seropositivity / Q000473:pathology*; D015497:HIV-1 / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000473:pathology*; D006801:Humans; D008969:Molecular Sequence Data; D009000:Monocytes / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D012326:RNA Splicing; D013601:T-Lymphocytes / Q000502:physiology
D015703:Antigens, CD; D018952:Antigens, CD34; D016376:Oligonucleotides, Antisense