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Efficient transformation of previously activated and dividing T lymphocytes
by human T cell leukemia-lymphoma virus
S Merl, B Kloster, J Moore, C Hubbell, R Tomar, F Davey, D Kalinowski, A Planas, G Ehrlich and D Clark
Modifying previously reported techniques, we attempted to increase the
efficiency of human T cell leukemia-lymphoma virus (HTLV) transformation of
human T lymphocytes. Lethally irradiated donor cells (DCs) were cultured
with target mononuclear cells (TMCs). DCs included ten HTLV+ T cell lines
with varying degrees of virus expression or seven cell lines that do not
express HTLV. TMCs were prepared from 20 cord and 16 adult peripheral blood
samples, including eight patients with acquired immunodeficiency syndrome
(AIDS). TMCs were either added directly to the DCs or were first stimulated
with phytohemagglutinin (PHA) (5 micrograms/mL) and grown in T cell growth
factor (TCGF) prior to exposure to DCs. The presence of integrated HTLV
proviral DNA in the transformed cells was determined by dot blot
hybridization, utilizing a cloned probe to the HTLV-I genome. HTLV
production by transformed TMCs was assessed for HTLV p19, reverse
transcriptase, and virus particles. No transformation occurred with T cell
donor lines that do not express HTLV. Low virus expressor DCs could only,
with rare exception, transform preactivated TMCs. High-titer
virus-producing DCs could transform activated and nonactivated cord blood
cells and activated adult TMCs. Only MT-2 could routinely transform
nonactivated normal adult and activated AIDS TMCs. HUT 102 B2 could
transform only one activated AIDS sample, the cells of which initially
expressed HTLV-like proteins and virions. Transformed cell lines contained
subsets of mature T lymphocytes with variable HTLV expression. Prior
activation and culture of the T lymphocytes increases the probability and
rate of transformation by HTLV, allowing for biologic detection of low
HTLV- producing cells and for in vitro expansion of T lymphocyte subsets
from selected patients.
Volume 64,
Issue 5,
pp. 967-974,
11/01/1984
Copyright © 1984 by The American Society of Hematology
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