Infection of peripheral blood mononuclear cells and cell lines by cell-free human T-cell lymphoma/leukemia virus type I.
Previous studies of in vitro infection by human T-cell lymphoma/leukemia virus type I (HTLV-I) have required cocultivation of target cells with HTLV-I cell lines or vesicular stomatitis virus pseudotypes containing HTLV-I envelope proteins. We report here the development of a cell-free infection assay for HTLV-I. Target cells were incubated with purified, DNase-treated HTLV-I virions for 4 h at 37 degrees C. Target cell DNA was then analyzed for the presence of newly synthesized HTLV-I proviral DNA by the highly sensitive polymerase chain reaction. Using this assay system, we have been able to consistently detect in vitro infection of a variety of cellular targets by different HTLV-I isolates. Optimal infection required the presence of 10 micrograms of DEAE-dextran per ml. The assay was dose dependent with respect to virus input. In general, the amount of proviral DNA detected correlated with the level of HTLV-I receptors present on the surface of the target cells, as measured by fluorochrome-labelled HTLV-I binding. Finally, the specificity of the assay was confirmed by demonstrating that the cell line, L1q, a somatic cell hybrid containing human chromosome 17q, to which the gene for the HTLV-I receptor has been mapped, was susceptible to infection by HTLV-I, while the parental mouse cell line from which it was derived, LMTK-, which lacks human chromosome 17q, was not.
Fan
N
N
Department of Microbiology and Immunology, SUNY Health Science Center, Syracuse.
Gavalchin
J
J
Paul
B
B
Wells
K H
KH
Lane
M J
MJ
Poiesz
B J
BJ
D000328:Adult; D002460:Cell Line; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D015490:HTLV-I Infections / Q000209:etiology*; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014773:Virology / Q000379:methods
D004279:DNA, Viral; D014764:Viral Proteins