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Forschungsdatenbank PMU-SQQUID

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.
SunderPlassmann, R; Breiteneder, H; Zimmermann, K; Strunk, D; Majdic, O; Knapp, W; Holter, W
Blood. 1996; 87(12):5179-5184
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Strunk Dirk

Abstract

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.


Useful keywords (using NLM MeSH Indexing)

Antibodies, Monoclonal/immunology

Antibodies, Monoclonal/pharmacology*

Antigens, CD28/immunology

Base Sequence

CD4-Positive T-Lymphocytes/drug effects

CD4-Positive T-Lymphocytes/immunology*

Cell Differentiation/drug effects

Cell Division/drug effects

Cells, Cultured

Clone Cells

Cytokines/immunology

Cytokines/pharmacology*

Gene Expression Regulation/drug effects

Gene Rearrangement, beta-Chain T-Cell Antigen Receptor

Humans

Interferon-gamma/biosynthesis

Interferon-gamma/pharmacology

Interleukin-1/pharmacology

Interleukin-2/biosynthesis*

Interleukin-2/genetics

Interleukin-2/pharmacology

Interleukin-4/biosynthesis

Interleukin-4/pharmacology

Interleukin-6/pharmacology

Lymphocyte Activation/drug effects*

Molecular Sequence Data

Polymerase Chain Reaction

Receptor-CD3 Complex, Antigen, T-Cell/immunology

Th1 Cells/cytology

Th2 Cells/cytology