Immunology Letters, 41 (1994) 13-17
0165 - 2478 / 94 / $ 7.00 © 1994 Elsevier Science B.V. All rights reserved
IMLET 02116
Discrepancy between direct and antibody-dependent cytotoxic
activities of human LAK cells
Michael P. Potapnev a'*, Tatyana S. Garbuzenco a, Natalia V. Goncharova a, Vladimir D.
Zobnin a, Oleg V. Shadrin b and Svetlana N. Bykovskaya b
~Institute of Hematology and Blood Transfusion, Minsk 223059, Belarus; bCancer Research Centre, Moscow 115478, Russia
(Received 20 January 1994; accepted 16 February 1994)
1. Summary
Human lymphokine-activated killer (LAK) cells
display cytotoxic activity against natural killer
(NK)-resistant tumor cells in an antibody-indepen-
dent and -dependent manner. We compared LAK
cell-mediated antibody-independent cytotoxicity
(LAK activity) and antibody-dependent cellular cyto-
toxicity (ADCC) against untreated and antibody-
coated Raji cells, respectively. Human lymphocytes
showed drastically increased LAK activity after sti-
mulation with interleukin-2 (IL-2) for 3 or 7 days
when compared to non-activated cells. The level of
ADCC was reduced for 3-day-generated LAK cells
and augmented for 7-day-generated LAK cells as
compared to non-activated cultured lymphocytes.
Phenotypical analysis revealed IL-2-induced up-regu-
lation of the proportion of CDIIb + (but not
CD16 +) lymphocyte subpopulation in 7-day-gener-
ated LAK cells. The data imply that human LAK
cells exhibit antibody-dependent and -independent
cytotoxic activities via distinct effector pathways at
different stages of generation. These stages may be
associated with changes in adhesion molecule
(CD 1l b/CD 18) expression on the surface of IL-2-ac-
tivated lymphocytes.
2. Introduction
Lymphokine-activated killer (LAK) cells have
Key words: Lymphokine-activated killer cell; Phenotyping; Direct
cytotoxicity; Antibody-dependent cellular cytotoxicity; (Human)
*Corresponding author: Michael P.Potapnev, Institute of Hematol-
ogy and Blood Transfusion, Dolginovsky tr. 160, 223059, Minsk,
Belarus.
been shown to exert strong anti-tumor activity in
vivo and in vitro [1,2]. Generation of LAK cells in
culture of human peripheral blood lymphocytes acti-
vated with interleukin 2 (IL-2) for short (2-3 days) or
long (7-10 days) term may induce accordingly natur-
al killer (NK)-like or T-cell-like LAK cells as esti-
mated on the basis of phenotypical marker differ-
ences [3,4]. Nevertheless, direct cytotoxic activity of
LAK cells has been shown not to depend upon their
NK/T-cell-related surface markers [5]. LAK cells also
exert indirect, antibody-dependent cellular cytotoxi-
city (ADCC) after induction with IL-2 in vivo and
in vitro [6]. It is known that both resting and acti-
vated lymphocytes display ADCC and that FcTR-
bearing NK-like cells are the principal effector lym-
phocytes [7,8].
We have undertaken a comparative study of the
direct cytotoxic (LAK) and indirect (ADCC) activ-
ities of short- and long-term generated LAK cells.
We found that after IL-2 activation for 3 days or 7
days human lymphocytes exhibit a different profile of
LAK and ADCC activities and peculiarities in CD16
and CD1 lb/CD18 surface molecules expression may
be associated with these differences.
3. Materials and Methods
3.1. Cells
Mononuclear cells were isolated from fresh hepar-
inized donor blood by a standard Ficoll/Hypaque
centrifugation procedure. Purified lymphocytes were
obtained from mononuclear cells treated with carbo-
nyl iron (4.5-5.2 /tm, Sigma) to delete phagocytic
cells (monocytes). Immunofluorescent analysis of
purified lymphocytes indicated < 0.1% contamina-
SSDI 0165-2478(94)00032-M