Brunner Cornelia, Kreth Hans Wolfgang, Ochs Hans D, Schuster Volker
Children's Hospital, University of Würzburg, Germany.
J Clin Immunol. 2002 Jul;22(4):244-51. doi: 10.1023/a:1016097010274.
X-linked agammaglobulinemia (XLA) is caused by mutations in the gene encoding the cytoplasmic Bruton's tyrosine kinase (Btk). Btk has been shown to play an essential role in the development of B1 (CD5+) and conventional circulating mature B cells (B2) in mouse and man. It has been shown in earlier studies that Btk is involved in both the BCR- and CD40-mediated signaling pathways. In this study, we analyzed the responsiveness of Epstein-Barr virus (EBV) transformed B cells from nine XLA patients to CD40 stimulation, particularly the CD40 induced activation of c-Jun N-terminal kinase (JNK). In eight XLA patients the JNK activation was unimpaired and in one case INK could not be activated by anti-CD40 stimulation. Btk protein expression was detectable by Western blotting in six cases, in one case Btk expression was drastically reduced, and in three cases no Btk expression could be observed. Btk kinase activity was found in three cases and it was reduced in one and not detectable in five cases. Furthermore, in one female patient with an agammaglobulinemia, Btk expression and function as well as JNK activation by CD40 stimulation was unimpaired. Our findings demonstrate that INK activation via the CD40 signaling pathway is intact in EBV-transformed B cells of most if not all XLA patients, independent of the mutation and its effect on Btk expression and kinase activity. We suggest that Btk is not necessary for the activation of INK upon CD40 stimulation, at least in the B cell subpopulation we had studied. We cannot exclude that these B cells belong to a "leaky" B-cell subpopulation in which the CD40 signaling pathway has become independent of Btk function.
X连锁无丙种球蛋白血症(XLA)是由编码胞质布鲁顿酪氨酸激酶(Btk)的基因突变引起的。已证明Btk在小鼠和人类的B1(CD5 +)和传统循环成熟B细胞(B2)的发育中起重要作用。早期研究表明,Btk参与BCR和CD40介导的信号通路。在本研究中,我们分析了9名XLA患者的爱泼斯坦-巴尔病毒(EBV)转化B细胞对CD40刺激的反应性,特别是CD40诱导的c-Jun N末端激酶(JNK)激活。在8名XLA患者中,JNK激活未受损,在1例中,INK不能被抗CD40刺激激活。通过蛋白质印迹法在6例中可检测到Btk蛋白表达,在1例中Btk表达大幅降低,在3例中未观察到Btk表达。在3例中发现了Btk激酶活性,在1例中降低,在5例中未检测到。此外,在1例患有无丙种球蛋白血症的女性患者中,Btk表达和功能以及CD40刺激引起的JNK激活未受损。我们的研究结果表明,在大多数(如果不是全部)XLA患者的EBV转化B细胞中,通过CD40信号通路激活INK是完整的,与突变及其对Btk表达和激酶活性的影响无关。我们认为,至少在我们研究的B细胞亚群中,Btk对于CD40刺激后INK的激活不是必需的。我们不能排除这些B细胞属于“渗漏”B细胞亚群,其中CD40信号通路已变得独立于Btk功能。