M. Lucchiari et al., HUMAN IMMUNE-RESPONSE TO HIV-1-NEF .1. CD45RO(-) T-LYMPHOCYTES OF NONINFECTED DONORS CONTAIN CYTOTOXIC T-LYMPHOCYTE PRECURSORS AT HIGH-FREQUENCY, International immunology, 6(11), 1994, pp. 1739-1749
The immune response of peripheral blood lymphocytes (PBL) of non-expos
ed human individuals to the Nef protein of HIV-1 was studied. Nef is a
regulatory protein of HIV which is immediately expressed after infect
ion and which seems to be important in the pathogenicity of HIV. Nef m
ay therefore serve as a potential target for effective immunity agains
t HIV infection. Epstein-Barr (EBV)-transformed lymphoblastoid a cell
lines (LCL) were established from four healthy young seronegative adul
ts and transfected with the Nef gene. These cells served as stimulator
cells for autologous PBL in vitro and as target cells for CTL. CTL re
sponses were readily generated against Nef-transfected LCL, consisting
of Nef-specific and putative EBV-specific CTL. Nef-specific CTL were
generated exclusively from CD8(+) cells and were MHC class I restricte
d. Since a vigorous Nef-specific CTL response in non-infected individu
als was unexpected, CTL precursor frequencies were determined by limit
ing dilution analyses in non-fractionated PBL and in pal separated int
o the CD45RO(-) (naive) and CD45RO(+) (memory) T cell populations. As
expected, the putative EBV-specific CTL precursors were predominantly
found in the CD45RO(+) subset at frequencies typical for memory T cell
s. Nef-specific CTL precursors, in contrast, were found predominantly
in the CD45RO(-) population, at even higher frequencies of -1/1000-1/3
000. Nef may thus display either an unusually high number of immunogen
ic peptides or a limited number of peptides presented in a very effici
ent way, so that many T cells including low affinity cells, would be t
riggered.