title: Characterization of novel Bruton's tyrosine kinase gene mutations in Central European patients with agammaglobulinemia.
abstract: X-linked agammaglobulinemia (XLA) is an immunodeficiency disorder caused by mutations in the gene coding for Bruton's tyrosine kinase (BTK). In this study we investigated 10 male patients with XLA-compatible phenotype (agammaglobulinemia and undetectable B cells in peripheral blood) from 9 unrelated Central European families. We identified seven different mutations, six of which were novel. One previously described point mutation caused a premature stop codon (p.C464X), two point mutations resulted in amino acid exchanges (p.W588R; p.G419E), and two point mutations affected splice sites (c.305-1G>A; c.391+1G>A). We further detected one deletion (c.1921_1927del CGTCCCA) and one large duplication. The duplication resulted from Alu element-induced unequal homologous recombination, which was only detectable by extended analysis of cDNA, while direct sequencing of genomic DNA gave a false negative result. Western blot analysis revealed that the patients with the p.W588R and the p.G419E amino acid substitutions, respectively, produced full length BTK, but in clearly diminished amounts. The patient with the 7bp deletion expressed low amounts of protein which might represent truncated BTK. All other genomic alterations resulted in complete loss of BTK protein. In two patients from unrelated families BTK protein expression was normal and no Btk gene mutation was detected. The results of this study further substantiate the importance of using elaborate molecular analysis with different detection techniques to obtain an explicit molecular diagnosis in patients with suspected XLA.
authors.affiliation: Immunology Outpatient Clinic, Schwarzspanierstr. 15/1/9, 1090 Vienna, Austria.
authors.forename: Doris
authors.initials: D
authors.lastname: Kristufek
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Aspalter
authors.forename: Martha Marianne
authors.initials: MM
authors.lastname: Eibl
authors.forename: Hermann Maximilian
authors.initials: HM
authors.lastname: Wolf
mesh_terms: D000328:Adult; D000077329:Agammaglobulinaemia Tyrosine Kinase; D000361:Agammaglobulinemia / Q000175:diagnosis* / Q000235:genetics; D019943:Amino Acid Substitution; D001483:Base Sequence; D026481:Blotting, Far-Western; D002648:Child; D006801:Humans; D018655:Lymphocyte Count; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D011505:Protein-Tyrosine Kinases / Q000032:analysis* / Q000235:genetics*
chemical_list: D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human