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Comparison of Three Different Methods for Determining Cell Proliferation in Breast Cancer Cell Lines
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Proliferative assays for B cell function.

James J Mond1, Mark Brunswick

  • 1Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.

Current Protocols in Immunology
|April 25, 2008
PubMed
Summary
This summary is machine-generated.

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This study outlines methods for measuring B cell proliferation using polyclonal stimulating agents like anti-immunoglobulin (anti-Ig) antibodies and lipopolysaccharide (LPS). These mitogens activate most B cells, simplifying the assessment of their proliferative capacity.

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • B cell proliferation is crucial for adaptive immune responses.
  • Measuring B cell proliferation is essential for understanding immune function and disease.
  • Antigen-specific B cell isolation is labor-intensive, necessitating alternative methods.

Purpose of the Study:

  • To describe procedures for measuring the proliferative capacity of purified B cells.
  • To detail methods utilizing polyclonal stimulating agents (mitogens) for B cell activation.
  • To provide protocols for assessing B cell proliferation via [3H]thymidine incorporation.

Main Methods:

  • Utilizing polyclonal stimulating agents (mitogens) such as anti-Ig antibodies (anti-IgM or anti-IgD) and lipopolysaccharide (LPS).
  • Cross-linking the B cell antigen receptor (surface immunoglobulin, sIg) with anti-Ig antibodies to induce proliferation.

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  • Measuring cell division by assessing the incorporation of [3H]thymidine into newly synthesized DNA.

    • Main Results:

    • Polyclonal stimulating agents activate the majority of B cells, facilitating robust proliferation measurements.
    • Both anti-Ig antibodies and LPS effectively stimulate B cell proliferation.
    • The [3H]thymidine incorporation assay quantifies the proliferative response.

    Conclusions:

    • The described methods provide reliable protocols for assessing B cell proliferative capacity.
    • Using polyclonal mitogens offers an efficient alternative to antigen-specific B cell proliferation assays.
    • These procedures are valuable for immunological research and diagnostics.