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Related Experiment Videos

A sensitive and efficient induction system for murine IgE. Single cell analysis at the clonal level.

F Ledermann1, C Schlienger, K Wagner

  • 1Research Department, CIBA-GEIGY, Ltd., Basel, Switzerland.

Journal of Immunological Methods
|August 9, 1991
PubMed
Summary

A new culture system using CB5.1 feeder cells enhances immunoglobulin E (IgE) expression analysis in single B cells. This method improves the detection of IgE-secreting B cells, crucial for understanding allergic responses.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Analyzing immunoglobulin E (IgE) expression in single B cells is challenging.
  • Existing feeder cell systems have limitations in supporting IgE class switching.

Purpose of the Study:

  • To develop and characterize a novel culture system for analyzing IgE expression by single murine B cells.
  • To evaluate the efficacy of the CB5.1 stroma cell line as a feeder layer for IgE class switching.

Main Methods:

  • Utilized a CB5.1 stroma cell line as a feeder layer for lipopolysaccharide (LPS)-stimulated B cells.
  • Employed interleukin-4 (IL-4) to induce IgE class switching.
  • Developed a highly specific sandwich-enzyme-linked immunosorbent assay (ELISA) for IgE detection.

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Main Results:

  • CB5.1 feeder cells significantly enhanced the frequency of IgE-secreting B cells (2.5-fold increase) compared to thymocytes.
  • Optimal IgE switching occurred at lower IL-4 concentrations than IgG1 switching, differing from thymocyte cultures.
  • The system allowed for the detection of IgE production at the single-cell level within B cell clones.

Conclusions:

  • The CB5.1 cell line provides a superior and defined feeder system for studying IgE class switching in B cells.
  • This novel system facilitates the analysis of IgE-producing cells at the single-cell level.
  • The findings offer a valuable tool for immunological research, particularly in allergy and B cell differentiation studies.