Development of a rapid whole blood flow cytometry procedure for the diagnosis of X-linked hyper-IgM syndrome patients and carriers

Clin Immunol Immunopathol. 1997 Nov;85(2):172-81. doi: 10.1006/clin.1997.4422.


The CD40 ligand expressed on activated T cells plays a pivotal role in B cell proliferation and differentiation. Mutations in the CD40 ligand gene, which alter its expression on the surface of activated T cells, are associated with the X-linked form of Hyper-IgM syndrome (XHIM). A rapid and simple, three-color whole blood flow cytometry procedure was developed for maximal expression and detection of the CD40L on the surface of in vitro activated CD4+ T cells. Approximately 90% of in vitro activated CD4+ T cells obtained from healthy controls expressed the CD40L compared to only 5% of in vitro activated CD4+ T cells obtained from the XHIM patients. The CD40L was expressed on approximately 50% of the in vitro activated CD4+ T cells obtained from the mothers of XHIM patients, consistent with a diagnosis of their carrier status. This is the first report of a whole blood procedure adapted for routine clinical use which is able to detect abnormal CD40L expression in XHIM patients and carriers.

MeSH terms

  • Adult
  • CD3 Complex / analysis
  • CD40 Antigens / biosynthesis
  • CD40 Ligand
  • Carcinogens / pharmacology
  • Carrier State / diagnosis*
  • Female
  • Flow Cytometry*
  • Genetic Linkage
  • Humans
  • Hypergammaglobulinemia / diagnosis*
  • Hypergammaglobulinemia / genetics*
  • Immunoglobulin M* / blood
  • Ionomycin / pharmacology
  • Ionophores / pharmacology
  • Ligands
  • Lymphocyte Activation
  • Membrane Glycoproteins / biosynthesis
  • Membrane Glycoproteins / blood
  • Membrane Glycoproteins / physiology
  • Syndrome
  • T-Lymphocytes / immunology
  • Tetradecanoylphorbol Acetate / pharmacology
  • Up-Regulation
  • X Chromosome / genetics*


  • CD3 Complex
  • CD40 Antigens
  • Carcinogens
  • Immunoglobulin M
  • Ionophores
  • Ligands
  • Membrane Glycoproteins
  • CD40 Ligand
  • Ionomycin
  • Tetradecanoylphorbol Acetate