Parvovirus B19-induced perturbation of human megakaryocytopoiesis in vitro.
Parvovirus B19 infection leads to transient aplastic crises in individuals with chronic hemolytic anemias or immunodeficiency states. An additional unexplained sequela of B19 infection is thrombocytopenia. Because B19 is known to have a remarkable tropism for human erythropoietic elements, and is not known to replicate in nonerythroid cells, the etiology of this thrombocytopenia is uncertain. We sought to define the pathobiology of B19-associated thrombocytopenia by examining the role of B19 on in vitro megakaryocytopoiesis. B19 infection of normal human bone marrow cells significantly suppressed megakaryocyte (MK) colony formation compared with mock-infected cells. No such inhibition was observed with a nonpathogenic human parvovirus, the adeno-associated virus 2 (AAV). The B19-MK cell interaction was also studied at the molecular level. Whereas low-density bone marrow cells containing erythroid precursor cells supported B19 DNA replication, no viral DNA replication was observed in B19-infected MK-enriched fractions as determined by the presence of viral DNA replicative intermediates on Southern blots. However, analysis of total cytoplasmic RNA isolated from B19-infected MK fractions showed a low-level expression of the B19 genome as detected by quantitative RNA dot blots as well as by Northern analysis. Furthermore, a frame-shift mutation in a recombinant AAV-B19 hybrid genome segment that encodes the viral nonstructural (NS1) protein significantly reduced the observed inhibition of MK colony formation. These studies indicate tissue-tropism of B19 beyond the erythroid progenitor cell, and lend support to the hypothesis that B19 genome expression may be toxic to cell populations that are nonpermissive for viral DNA replication.
Srivastava
A
A
Department of Medicine, Walther Oncology Center, Indiana University School of Medicine, Indianapolis 46202-5120.
Bruno
E
E
Briddell
R
R
Cooper
R
R
Srivastava
C
C
van Besien
K
K
Hoffman
R
R
D015152:Blotting, Northern; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002213:Capsid / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000235:genetics; D006410:Hematopoiesis; D006801:Humans; D008533:Megakaryocytes / Q000382:microbiology / Q000502:physiology*; D010321:Parvoviridae / Q000235:genetics / Q000502:physiology*; D013921:Thrombocytopenia / Q000209:etiology / Q000473:pathology; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D017361:Viral Nonstructural Proteins; D014779:Virus Replication / Q000187:drug effects
D004279:DNA, Viral; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins