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.