Anti-CD3 + interleukin-2 stimulation of marrow and blood: comparison of
proliferation and cytotoxicity
PM Anderson, AC Ochoa, NK Ramsay, D Hasz and D Weisdorf
University of Minnesota Bone Marrow Transplant Program, Minneapolis 55455.
The proliferation and in vitro cytolytic activity of interleukin-2 (IL-
2)-activated and anti-CD3 + IL-2-stimulated marrow mononuclear cells (MMC)
and peripheral blood mononuclear cells (PBMC) were studied. Samples from 8
normal donors, 15 patients with acute lymphoblastic leukemia (ALL), and 7
patients with non-Hodgkin's lymphoma (NHL) in remission were cultured in
IL-2 (100 U/mL) or IL-2 (100 U/mL) plus anti- CD3 (10 ng/mL). MMC as well
as PBMC samples demonstrated significant synergy between IL-2 and anti-CD3
in the promotion of proliferation as measured by 3H thymidine incorporation
on day 5 (P less than .001) or fold increase in cell number on day 14.
Cryopreserved marrow specimens had equally rapid proliferation as fresh MMC
when cultured in the presence of anti-CD3 + IL-2. Anti-CD3 concentrations
of 3, 11, 33, and 100 ng/mL augmented proliferation similarly in the
presence of IL-2 (0.1 to 100 U/mL). Mean fold increases in cell number of
both marrow- and blood-derived cultures after 14 days were significantly
higher for anti-CD3 + IL-2-stimulated cultures compared with cultures
stimulated with IL-2 only (50- to 200-fold increase in cell number; P =
.01). Comparison of remission MMC and PBMC from ALL and NHL patients with
normal controls showed equivalent growth rates of activated cultures at 7,
14, and 21 days. Marrow purging with immunotoxin anti-CD19 pokeweed
antiviral protein plus 4HC had no significant effect on proliferation of
anti-CD3 + IL-2-stimulated MMC cultures in patients with ALL. Cytolytic
activity of IL-2- and IL-2 + anti-CD3-activated PBMC and MMC cultures was
assessed in 51Cr release assays using K562 (natural killer
([NK]-sensitive), Daudi (Burkitt's lymphoma-, NK-resistant), and Nalm-6
(ALL-, lymphokine-activated killer [LAK]-resistant) cell lines and
cryopreserved ALL blasts. Cytolytic activity on a per-cell basis (percent
cytotoxicity at an effector:target ratio of 30:1) was similar in
IL-2-activated PBMC- and MMC-derived cultures from ALL patients. MMC
activated with anti-CD3 plus IL-2 killed Daudi significantly less well than
IL-2-activated cultures on days 12 and 19 (P = .03); no significant
differences were observed in lysis of LAK-resistant Nalm-6 or cryopreserved
ALL blast targets. Dose response of anti-CD3 augmentation of Daudi and
Nalm-6 killing was different in IL-2- and IL- 2 + anti-CD3-stimulated
cultures.(ABSTRACT TRUNCATED AT 400 WORDS)
Volume 80,
Issue 7,
pp. 1846-1853,
10/01/1992
Copyright © 1992 by The American Society of Hematology
