Single human T cells stimulated in the absence of feeder cells transcribe
interleukin-2 and undergo long-term clonal growth in response to defined
monoclonal antibodies and cytokine stimulation
R Sunder-Plassmann, H Breiteneder, K Zimmermann, D Strunk, O Majdic, W Knapp and W Holter
Institute of Immunology, University of Vienna, Austria.
The two-signal model of T-cell activation postulates that T lymphocytes
require at least two distinct signals for activation. This model has been
established with bulk cultures of T cells in which T-cell-T-cell
interaction can occur, possibly delivering further unrecognized
costimulatory signals. The signal requirements of single T cells for the
induction of clonal cell growth or the transcription of cytokines would
best be studied in a cell cloning system in the absence of feeder cells;
however, such an experimental system has not been reported so far. In this
study, we report the long-term cloning of human resting peripheral blood
CD4+CD45RO- T cells under feeder cell-free conditions in response to CD3
and CD28 stimulation in the presence of exogenous interleukin-2 (IL-2).
Cloning efficiency ranged from 40% to 60% depending on the presence of
additional cytokines IL-1 and IL-6. Single- call polymerase chain reaction
showed that transcription of IL-2 occurred in cells stimulated through CD3
plus CD28 alone. T cells grown in response to CD3 plus CD28 plus IL-2
stimulation produced both IL-4 and interferon-gamma (IFN-gamma) on
restimulation (Th0 cells) and could be functionally differentiated into
Th1- or Th2-type cells by the addition of IFN-gamma or IL-4, respectively,
during cell cloning. These data show on the single-cell level a two-signal
model of T- cell activation for the transcription of IL-2. In addition,
these experiments show that IFN-gamma and IL-4 exert their T-cell-
differentiating effects directly on the T cell without any further need for
antigen-presenting cells. Together, our experiments show the feasability of
a defined long-term clonal cell culture system to study the growth and
differentiation of human T lymphocytes.
Volume 87,
Issue 12,
pp. 5179-5184,
06/15/1996
Copyright © 1996 by The American Society of Hematology
