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Human T hybridoma-derived immunoglobulin-binding factors
International Archives of Allergy and Applied Immunology
|January 1, 1987
Summary
Researchers identified novel immunoglobulin-binding factors from a unique human T hybridoma cell line. These factors inhibit immunoglobulin E synthesis and may play a role in immune regulation.
Area of Science:
- Immunology
- Cell Biology
Background:
- A human T hybridoma cell line was developed lacking low-affinity immunoglobulin E (IgE) surface receptors.
- This cell line spontaneously produces immunoglobulin-binding factors.
Purpose of the Study:
- To characterize the immunoglobulin-binding factors produced by the T hybridoma.
- To investigate the biological activities of these factors, particularly their role in IgE and IgG synthesis.
Main Methods:
- Purification of binding factors using concanavalin A affinity chromatography, IgE immunoaffinity column, and SDS-PAGE.
- Western blot analysis to assess glycosylation and immunoglobulin-binding capabilities.
- Assay of biological activity on U266 B cell line and B cells from atopic donors.
Main Results:
- Purification yielded five major protein bands (13, 15, 30, 60, <150 kDa), all variably glycosylated and capable of binding IgE.
- A 13 kDa peptide inhibited IgE synthesis in a B cell line and in B cells from atopic donors.
- Semipurified factors inhibited Pokeweed mitogen-induced IgG synthesis while stimulating IgE synthesis.
Conclusions:
- The identified factors exhibit broad immunoglobulin-binding properties and modulate B cell Ig synthesis.
- Glycosylation is not essential for immunoglobulin binding but may influence biological activity.
- The exact molecular nature of the different protein entities remains to be elucidated.