title: Development of a rapid whole blood flow cytometry procedure for the diagnosis of X-linked hyper-IgM syndrome patients and carriers.
abstract: 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.
authors.affiliation: Children's Memorial Hospital, Northwestern University, Chicago, Illinois 60614, USA. mogorman@nwu.edu
authors.forename: M R
authors.initials: MR
authors.lastname: O'Gorman
authors.forename: D
authors.initials: D
authors.lastname: Zaas
authors.forename: M
authors.initials: M
authors.lastname: Paniagua
authors.forename: V
authors.initials: V
authors.lastname: Corrochano
authors.forename: P R
authors.initials: PR
authors.lastname: Scholl
authors.forename: L M
authors.initials: LM
authors.lastname: Pachman
mesh_terms: D000328:Adult; D017252:CD3 Complex / Q000032:analysis; D019013:CD40 Antigens / Q000096:biosynthesis; D023201:CD40 Ligand; D002273:Carcinogens / Q000494:pharmacology; D002353:Carrier State / Q000175:diagnosis*; D005260:Female; D005434:Flow Cytometry; D008040:Genetic Linkage; D006801:Humans; D006942:Hypergammaglobulinemia / Q000175:diagnosis* / Q000235:genetics*; D007075:Immunoglobulin M / Q000097:blood; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008024:Ligands; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000097:blood / Q000502:physiology; D013577:Syndrome; D013601:T-Lymphocytes / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015854:Up-Regulation; D014960:X Chromosome / Q000235:genetics*
chemical_list: D017252:CD3 Complex; D019013:CD40 Antigens; D002273:Carcinogens; D007075:Immunoglobulin M; D007476:Ionophores; D008024:Ligands; D008562:Membrane Glycoproteins; D023201:CD40 Ligand; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate