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Surface phenotypes of human T cell lines generated after WGA- and PHA-stimulation
J K Siwicki1, H Skurzak, J A Steffen
1Department of Immunology, Maria Skłodowska-Curie Memorial Cancer Center, Warsaw, Poland.
Archivum Immunologiae Et Therapiae Experimentalis
|March 25, 1998
Summary
Human T lymphocytes can be cultured long-term using IL-2 and stimulation. These cultures yield diverse T cell receptor alpha beta (TcR αβ) cells, showing long-term growth potential isn't limited to one subset.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Human T lymphocytes from blood or spleens can be propagated in long-term culture with Interleukin-2 (IL-2).
- Previous methods involved stimulation with wheat germ agglutinin (WGA) or phytohemagglutinin (PHA) and repeated restimulation.
Purpose of the Study:
- To characterize the cell types and phenotypes within long-term cultured human T lymphoblast lines.
- To investigate the potential for selective outgrowth of specific T cell subsets during extended culture.
Main Methods:
- Generation of lymphoblast lines from normal human T lymphocytes using IL-2 and WGA or PHA stimulation.
- Analysis of cell surface phenotypes, including CD3, T cell receptor alpha beta (TcR αβ), CD4, and CD8 expression.
- Monitoring cell populations over extended culture periods (over 100 population doublings).
Main Results:
- Lymphoblast lines predominantly expressed the CD3 molecule and the T cell receptor alpha beta (TcR αβ) heterodimer.
- PHA-initiated lines showed a prevalence of CD4-CD8+ cells.
- WGA-initiated lines exhibited varied proportions of CD4+CD8-, CD4-CD8+, and CD4-CD8- cells.
- Some lines demonstrated selective outgrowth of TcR αβ CD4+CD8- or CD4-CD8- cells over time.
Conclusions:
- Long-term growth potential of human normal T lymphoblasts is not restricted to a single subset.
- Surface phenotypes acquired during T cell differentiation can be maintained through extensive in vitro culture (over 100 population doublings).