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Manipulation of human B cells to confer immortality
1SmithKline Beecham Pharmaceuticals, Department of Cellular Biochemistry and Immunology, King of Prussia, PA 19406-2799.
Human Antibodies and Hybridomas
|October 1, 1992
Summary
Electroporation effectively immortalizes human lymphocytes, creating stable cell lines for studying B cell differentiation and antibody production. This method yields valuable resources for immunological research.
Area of Science:
- Immunology
- Cell Biology
Background:
- Lymphocytes are crucial for immune responses but are difficult to immortalize for research.
- Existing methods for lymphocyte immortalization have limitations.
Purpose of the Study:
- To establish a method for immortalizing human lymphocytes at various differentiation stages using electroporation.
- To characterize the resulting immortalized B cell lines for stability and functionality.
Main Methods:
- Electroporation was used to introduce tumor-derived genomic DNA into activated human peripheral blood lymphocytes.
- Transfected cell clones were selected and analyzed for phenotype, growth rate, morphology, and antibody production.
- Karyotype analysis was performed to assess chromosomal stability.
Main Results:
- Activation of lymphocytes was essential for efficient immortalization.
- A panel of immortalized human B cell lines representing different differentiation stages was generated.
- Transfectants exhibited stable morphology, growth, and immunoglobulin production (up to 10 µg/mL).
- Chromosome analysis revealed normal karyotypes in the immortalized cells.
Conclusions:
- Electroporation is an effective technique for generating immortalized human lymphocytes.
- The resulting cell lines are stable and represent various B cell differentiation stages.
- These immortalized B cells serve as a valuable resource for studying B cell biology, differentiation, and immunoglobulin production.