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Related Experiment Videos

Absolute TCD4+ counting by a minimalist dual-platform flow cytometric method.

R Chianese1, E Nebuloni, M De Paschale

  • 1Department of Hematology and Blood Transfusion, Hospital of Legnano, Milano, Italy. chianese.r@libero.it

Journal of Biological Regulators and Homeostatic Agents
|April 7, 2004
PubMed
Summary

A new flow cytometry method using total white blood cell (WBC) gating for absolute TCD4+ counting showed fair agreement with the standard lymphocyte gating method. This WBC gating approach yields systematically higher TCD4+ counts, potentially due to lower lymphocyte percentage estimations by hematological analyzers.

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Area of Science:

  • Immunology
  • Hematology
  • Clinical Chemistry

Background:

  • Accurate TCD4+ cell enumeration is crucial for monitoring HIV infection and guiding treatment.
  • Current flow cytometry methods rely on lymphocyte gating, which can be influenced by variations in lymphocyte percentages.
  • Developing a simplified and robust TCD4+ counting strategy is essential for resource-limited settings.

Purpose of the Study:

  • To compare the performance of an absolute TCD4+ counting method using total white blood cell (WBC) gating against the standard lymphocyte (Ly) gating method.
  • To evaluate the feasibility of a flow cytometric (FCM) minimalist strategy for TCD4+ enumeration.

Main Methods:

  • Analysis of 132 routine peripheral blood samples from HIV-infected patients using flow cytometry.

Related Experiment Videos

  • Comparison of two gating strategies: standard Ly gating (CD45++SSClow) and total WBC gating (CD45/CD4 biparametric scatter).
  • Calculation of absolute TCD4+ counts using both lymphocyte and WBC counts from a hematological analyzer (HA).

    • Main Results:

    • A fair agreement was observed between the TCD4+ counts obtained by the total WBC gate and the standard Ly gate (r2 = 0.9847).
    • The total WBC gate method resulted in systematically higher TCD4+ counts compared to the standard method (mean difference = -56.22).
    • Lymphocyte counts also showed good agreement between methods (r2 = 0.9764), but with a larger mean difference (-362.93).

    Conclusions:

    • The total WBC gating method offers a viable alternative for TCD4+ enumeration with fair agreement to the standard method.
    • The observed higher TCD4+ counts with WBC gating may be linked to systematic underestimation of lymphocyte percentages by hematological analyzers.
    • This FCM minimalist strategy warrants further investigation for its clinical utility in TCD4+ counting.