Stromal cell lines derived from LP-BM5 murine leukemia virus-infected long-term bone marrow cultures impair hematopoiesis in vitro.
Murine acquired immunodeficiency syndrome (MAIDS) induced by defective LP-BM5 murine leukemia virus is a disease with many similarities to human AIDS. Previous studies indicated that the depressed hematopoiesis observed in LP-BM5-infected marrow cultures may be attributable to a defect of hematopoietic stroma. We report here the generation of permanent stromal cell lines from noninfected and LP-BM5-infected marrow cultures. Retrovirus infection was confirmed by polymerase chain reaction for viral genome. The ability of these cell lines to support in vitro hematopoiesis was studied. Results indicated that, when cocultured with normal or infected nonadherent mononuclear cells, noninfected cell lines efficiently supported the production of hematopoietic precursors, whereas viral-infected cell lines induced suppression of both normal and viral-infected progenitors. Expression of cytokine genes in stromal cell lines was also examined. All cell lines expressed equivalent levels of transcripts for stem cell factor and tumor necrosis factor alpha. However, infection was associated with higher levels of interleukin-4 and transforming growth factor beta 1 transcript expression. These findings suggest that infected stromal cell lines exhibit a defective hematopoietic microenvironment that produced altered cytokine expression resulting in faulty hematopoiesis. Further characterization of the defective cell lines should prove valuable for studies of the pathogenesis of murine AIDS.
Tse
K F
KF
Department of Internal Medicine, University of Kentucky Medical Center, Lexington.
Morrow
J K
JK
Hughes
N K
NK
Gallicchio
V S
VS
D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000473:pathology*; D002460:Cell Line; D002469:Cell Separation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D016207:Cytokines / Q000096:biosynthesis*; D017931:DNA Primers; D003673:Defective Viruses / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D015870:Gene Expression; D015750:Genes, gag; D006410:Hematopoiesis; D016298:Hematopoietic Cell Growth Factors / Q000096:biosynthesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000473:pathology* / Q000502:physiology; D007377:Interleukin-3 / Q000096:biosynthesis; D015847:Interleukin-4 / Q000096:biosynthesis; D009052:Leukemia Virus, Murine / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016183:Murine Acquired Immunodeficiency Syndrome / Q000382:microbiology; D009685:Nuclear Envelope / Q000382:microbiology / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D019089:Stem Cell Factor; D016212:Transforming Growth Factor beta / Q000096:biosynthesis; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D016207:Cytokines; D017931:DNA Primers; D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D019089:Stem Cell Factor; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D015847:Interleukin-4