Rapid whole blood flow cytometric test to detect ICOS deficiency in patients with common variable immunodeficiency

Int Arch Allergy Immunol. 2006;140(4):342-4. doi: 10.1159/000093770. Epub 2006 Jun 2.

Abstract

Background/aims: Common variable immunodeficiency (CVID) is the most common primary immunodeficiency. With respect to underlying defects it comprises a heterogeneous group of deficiencies. For some patients, distinct phenotypical abnormalities have been described, e.g. partial CD40L deficiency or complete ICOS deficiency. For the diagnosis of CD40L deficiency, a rapid whole blood flow cytometric method has been described several years ago. We aimed to determine if the same method can be used to diagnose ICOS deficiency.

Methods: Whole blood from 8 healthy volunteers was stimulated for 4 and 20 h with phorbol 12-myristate 13-acetate (PMA) and ionomycin. Induction of ICOS expression was analyzed on CD8-CD3+ lymphocytes using three-color flow cytometry. Blood from a patient with diagnosed ICOS deficiency was also analyzed.

Results: Whole-blood stimulation with PMA and ionomycin for 20 h resulted in a significant induction of ICOS expression on CD8-CD3+ lymphocytes in healthy volunteers. Four-hour incubation also demonstrated ICOS upregulation but to a much lower extent. In CD8-CD3+ lymphocytes from an ICOS-deficient patient, no ICOS expression could be induced following 20 h of stimulation.

Conclusion: ICOS expression can be analyzed using a rapid whole blood flow cytometric test.

MeSH terms

  • Antigens, CD / blood
  • Antigens, Differentiation, T-Lymphocyte / blood*
  • Antigens, Differentiation, T-Lymphocyte / metabolism
  • CD3 Complex / blood
  • CD8-Positive T-Lymphocytes / chemistry
  • CD8-Positive T-Lymphocytes / drug effects
  • CD8-Positive T-Lymphocytes / metabolism
  • Common Variable Immunodeficiency / blood*
  • Common Variable Immunodeficiency / pathology
  • Flow Cytometry / methods*
  • Humans
  • Inducible T-Cell Co-Stimulator Protein
  • Ionomycin / pharmacology
  • Lectins, C-Type
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • Antigens, CD
  • Antigens, Differentiation, T-Lymphocyte
  • CD3 Complex
  • CD69 antigen
  • ICOS protein, human
  • Inducible T-Cell Co-Stimulator Protein
  • Lectins, C-Type
  • Ionomycin
  • Tetradecanoylphorbol Acetate