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Ricin-resistant human T-cell hybridomas producing interferon gamma
Abstract:
Ricin-resistant variants of the SH9 T-cell line were selected after growth of this line in medium containing toxic amounts of ricin, a lectin derived from Ricinus communis. The ricin-resistant SH9 lines, SH9.R0 and SH9.R1, were demonstrated to be deficient in cell surface ricin-binding sites, but otherwise had the cellular phenotype of SH9 cells. Ricin-resistant T-cell hybridomas were prepared by fusion of SH9.R0 and SH9.R1 with activated T-lymphocytes. The presence of ricin in the selection medium rapidly killed unfused T-lymphocytes and prevented cell transformation by human T-cell leukaemia virus type 1 (HTLV-1) which is shed by the SH9.R0 and SH9.R1 cells. This ensured that the cells growing out were indeed hybridomas. Ricin-resistant T-cell hybridomas were characterised and also shown to lack cell surface receptors for ricin. Analysis of T-cell surface markers indicated that the T-cell hybridomas could be the result of fusions between SH9.R1 cells and T-helper lymphocytes or T-suppressor lymphocytes. All of the T-cell hybridomas prepared in this study spontaneously produced interferon gamma (IFN gamma).
Insights
Researchers developed ricin-resistant T-cell hybridomas by selecting for resistance to ricin, a plant toxin. These hybridomas lack ricin-binding sites and produce interferon gamma, offering a tool for immunological studies.
Area of Science:
- Immunology
- Cell Biology
- Toxicology
Background:
- SH9 T-cell line was used to generate ricin-resistant variants.
- Ricin is a toxic lectin from Ricinus communis.
- Cell surface ricin-binding sites are crucial for ricin toxicity.
Purpose of the Study:
- To create and characterize ricin-resistant T-cell hybridomas.
- To investigate the phenotype of ricin-resistant T-cells.
- To utilize these hybridomas for immunological research.
Main Methods:
- Selection of ricin-resistant SH9 T-cell lines (SH9.R0, SH9.R1).
- Fusion of ricin-resistant lines with activated T-lymphocytes.
- Characterization of resulting T-cell hybridomas, including surface marker analysis and interferon gamma production.
Main Results:
- Ricin-resistant SH9 lines (SH9.R0, SH9.R1) lacked cell surface ricin-binding sites.
- Ricin-resistant T-cell hybridomas were successfully generated.
- These hybridomas lacked ricin receptors and produced interferon gamma.
- Analysis suggested fusion with T-helper or T-suppressor lymphocytes.
Conclusions:
- Ricin resistance in SH9 cells is associated with a deficiency in ricin-binding sites.
- Ricin-resistant T-cell hybridomas can be generated and are useful research tools.
- These hybridomas spontaneously produce interferon gamma, indicating potential for immune modulation studies.