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Disease-causing STAT3 variants can be discriminated by a functional flow cytometry test.

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A new flow cytometry test accurately distinguishes STAT3 variants (dominant negative, gain-of-function, haploinsufficiency) in patients. This method simplifies the functional assessment of STAT3 variants, aiding in diagnosing inborn errors of immunity.

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STAT3classificationclinical laboratorydiagnosis

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Area of Science:

  • Immunology
  • Genetics
  • Molecular Biology

Background:

  • STAT3 variants are linked to various inborn errors of immunity through dominant negative (DN), gain-of-function (GOF), or haploinsufficiency (HI) mechanisms.
  • Assessing the biological impact of STAT3 variants is complex, often requiring extensive experimental procedures.

Purpose of the Study:

  • To develop a single, reliable functional test for STAT3 variants in patient primary cells.
  • To simplify the characterization of disease-causing STAT3 variants.

Main Methods:

  • Recruited patients with validated STAT3-DN, STAT3-GOF, and STAT3-HI variants.
  • Evaluated the IL-10-mediated inhibition ratio of TNF-α production in peripheral blood mononuclear cells (PBMC) using flow cytometry.
  • Optimized the assay and defined thresholds using ROC-AUC analysis.

Main Results:

  • STAT3-DN and STAT3-HI variants showed significantly diminished TNF-α inhibition, while STAT3-GOF variants showed augmented inhibition compared to healthy controls.
  • The optimized assay demonstrated high accuracy in discriminating between STAT3 variant types and healthy controls (ROC-AUC up to 1).

Conclusions:

  • The IL-10 mediated, STAT3-dependent TNFα inhibition ratio assay in PBMC-derived monocytes is a sensitive and specific method.
  • This flow cytometry-based test effectively assesses and differentiates STAT3-DN, -GOF, and -HI variants for research and clinical use.