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A deletion in the proximal untranslated pX region of human T-cell leukemia
virus type II decreases viral replication but not infectivity in vivo
GL Cockerell, J Rovnak, PL Green and IS Chen
Department of Pathology, College of Veterinary Medicine and Biomedical
Sciences, Colorado State University, Fort Collins 80523-1671, USA.
The function of untranslated (UT) nucleotide sequences in the proximal
portion of the pX region of the human T-cell leukemia virus (HTLV) family
of retroviruses remains enigmatic. Previous studies have shown that these
sequences are not necessary for the expression of viral proteins or for the
induction, transmission, or maintenance of the transformed cell type in
vitro. To determine the effect of the UT region in vivo, separate groups of
rabbits were inoculated with lethally irradiated, stable clones of the
human B-lymphoblastoid cell line, 729, transfected with either a
full-length wild-type HTLV-II clone (pH6neo) or a mutant clone containing a
324-bp deletion in the proximal UT portion of pX (pH6neo delta
UT[6661-6984]), or nontransfected 729 cells. All rabbits inoculated with
either wild-type or pX-deleted HTLV-II developed a similar profile and
titer of serum antibodies against HTLV-II antigens, as determined by
Western immunoblots, by 4 weeks postinoculation (PI). Antibody titers, as
determined by enzyme immunoassay, were similar between the two groups of
rabbits and increased over the 18-week period of study. All rabbits were
killed at 18 weeks PI, and spleen, peripheral blood lymphocytes (PBMC),
bone marrow, and mesenteric lymph node were assayed for HTLV-II tax/rex
sequences by quantitative polymerase chain reaction. Virus was detected in
all tissues tested from all rabbits inoculated with 729pH6neo cells
containing wild-type HTLV-II, which contained between 1.4 and 0.3 mean
copies of provirus per cell. In contrast, the distribution and number of
provirus copies were more limited in rabbits inoculated with 729pH6neo
delta UT(6661-6984) cells containing UT- deleted HTLV-II; in most tissues,
there was a fivefold to sevenfold reduction in mean provirus copies per
cell as compared with rabbits inoculated with wild-type HTLV-II. All
rabbits inoculated with control 729 cells remained negative for HTLV-II
infection, as determined by the same techniques. It was concluded that UT
sequences in the proximal portion of HTLV-II are not necessary for
infection but confer increased replicative capacity in vivo.
Volume 87,
Issue 3,
pp. 1030-1035,
02/01/1996
Copyright © 1996 by The American Society of Hematology
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