Bs. Weeks et al., NEUROTOXICITY OF THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 TAT TRANSACTIVATOR TO PC12 CELLS REQUIRES THE TAT AMINO-ACID-49-58 BASIC DOMAIN, Journal of neuroscience research, 42(1), 1995, pp. 34-40
The acquired immunodeficiency syndrome (AIDS) frequently involves the
central nervous system (CNS) and manifests as dementia due to encephal
itis or diffuse neurodegeneration. Human immunodeficiency virus type 1
(HIV-1) proteins, potentially transported into the CNS by mononuclear
inflammatory cells, have been implicated in the etiology of this HIV-
1 associated neurological dysfunction, Here we investigate the neuroto
xicity of the essential HIV-1 regulator protein Tat in vivo after micr
oinfusion into the rat brain and in vitro using PC12, NG108-15, and GT
17 neuronal cell lines, Infusion of either chemically synthesized Tat
(Tat86) or recombinant Tat (rTat) into the striatal gray matter in Spr
ague-Dawley rats resulted in postural deviation ipsilateral to the inf
usion, a clinical presentation in rats associated with complete striat
al dysfunction, Histologic examination 3 days after infusion revealed
massive necrosis in the area of the distribution of the infusion, Infu
sion of heat denatured rTat, peptide Tat49-58, or peptide Tat57-86 did
not result in clinically or histologically detectable brain damage, A
fter 3 days incubation in vitro, the lethal dose for half (LD50) of PC
12 cells due to rTat was 5 mu g/ml, The LD50 for Tat86 under the same
conditions was 10 mu g/ml. Tat49-58 and Tat57-86 peptides were not tox
ic in vitro even at 10-fold higher doses, At 5 mu g/ml, rTat was toxic
to 100% of GT17 cells after 24 hr, At 5 mu g/ml, Tat86 was toxic to 9
0% of the NG108-15 cells after 7 days of treatment, Prior experiments
have shown Tat49-58 is specifically recognized by a cell surface prote
in that mediates Tat uptake. Here we show that Tat toxicity is inhibit
ed by cotreatment with excess Tat49-58, suggesting Tat neurotoxicity r
equires binding to its surface ligand, Chloroquine, which increases nu
clear accumulation of Tat, enhances Tat toxicity to PC12 cells, sugges
ting that Tat internalization is a required step in the mechanism of i
ts toxicity. (C) 1995 Wiley-Liss, Inc.