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Related Experiment Videos

Quantitative analysis of lymphocyte CD11a using standardized flow cytometry

M Pallis1, A Robins, R Powell

  • 1Department of Immunology, University Hospital, Nottingham, UK.

Scandinavian Journal of Immunology
|December 1, 1993
PubMed
Summary
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This study refines lymphocyte CD11a density measurement using FACScan and standardized antibodies. Results show age-related CD11a expression and a reliable cutoff for CD8+ cells, enabling detection of abnormal immune cell groups.

Area of Science:

  • Immunology
  • Cell Biology
  • Flow Cytometry

Background:

  • CD11a is the alpha-subunit of leukocyte function-associated antigen-1 (LFA-1), a key molecule in leukocyte adhesion and costimulation.
  • Accurate measurement of cell surface molecule density is crucial for understanding immune cell function and identifying disease states.

Purpose of the Study:

  • To refine and validate a method for measuring lymphocyte CD11a density using FACScan.
  • To establish age-related reference ranges for CD11a expression on CD4+ and CD8+ lymphocytes.
  • To assess the method's precision and statistical power for detecting variations in CD11a expression.

Main Methods:

  • Utilized a FACScan flow cytometer with standardized fluorescent beads and a fluorescein-conjugated anti-CD11a antibody.
  • Measured CD11a density on peripheral blood CD4+ and CD8+ lymphocytes from 60 healthy individuals.

Related Experiment Videos

  • Established a cutoff point for CD11a expression on CD8+ cells to differentiate between low- and high-expressing subsets.

    • Main Results:

    • Demonstrated a linear correlation between age and mean CD11a density on both CD4+ (r=0.47) and CD8+ (r=0.71) lymphocytes.
    • Identified a reliable cutoff at 4.3 x 10^3 bound antibody molecules for CD8+ cells.
    • The method showed high statistical power (>99%) and precision (<4% difference) for detecting variations in CD11a expression.

    Conclusions:

    • The refined FACScan method provides a precise and powerful tool for quantifying lymphocyte CD11a density.
    • Age-dependent variations in CD11a expression are significant and should be considered in comparative studies.
    • The method is suitable for detecting abnormal CD11a expression in patient groups, as suggested by preliminary findings in infectious mononucleosis.