Smith C I, Baskin B, Humire-Greiff P, Zhou J N, Olsson P G, Maniar H S, Kjellén P, Lambris J D, Christensson B, Hammarström L
Center for BioTechnology, Karolinska Institute, NOVUM, Huddinge, Sweden.
J Immunol. 1994 Jan 15;152(2):557-65.
The gene mutated in the human disease, X-linked agammaglobulinemia (XLA), is related to the Src gene family of cytoplasmic protein-tyrosine kinases and is designated Btk (Bruton's agammaglobulinemia tyrosine kinase; formerly Atk/Bpk; the human gene is denoted BTK, using capital letters according to the kinase nomenclature). We have recently reported that this gene is expressed in B lymphocytes and that the specific mRNA was undetectable in T cells using Northern blotting. Further analyses of different sources of B and T lymphocytes confirmed this pattern. However, BTK transcripts were undetectable in four plasmacytoma lines. Moreover, as virtually normal amounts of BTK transcripts were found in PBMC from two patients carrying a point mutation in BTK, despite low B cell numbers, we anticipated that the gene would also be expressed in cells of other lineages. The erythroleukemia cell line K-562, the promyelocytic line HL-60 and the histiocytic lymphoma line U-937 were found to have BTK mRNA levels comparable to B cells. BTK mRNA was also detected in monocytes from healthy donors as well as in the human immature basophilic cell line KU812, in the human mast cell leukemia cell line HMC-1 and in the CD34 expressing myeloblast KG-1. A similar expression pattern was obtained when BTK protein was analyzed by immunoprecipitation and Western blotting. Using a polymerase chain reaction-based analysis, a small amount (less than 1% of the level in B cells) of BTK mRNA was identified in T lymphocytes. Our findings are compatible with a general expression of the BTK gene in hematopoietic cells, except in T lymphocytes and plasma cells, in which the transcript level is selectively down-regulated.
在人类疾病X连锁无丙种球蛋白血症(XLA)中发生突变的基因,与细胞质蛋白酪氨酸激酶的Src基因家族相关,被命名为Btk(布鲁顿无丙种球蛋白血症酪氨酸激酶;以前称为Atk/Bpk;根据激酶命名法,人类基因用大写字母表示为BTK)。我们最近报道,该基因在B淋巴细胞中表达,并且使用Northern印迹法在T细胞中未检测到特异性mRNA。对不同来源的B淋巴细胞和T淋巴细胞的进一步分析证实了这种模式。然而,在四种浆细胞瘤细胞系中未检测到BTK转录本。此外,尽管两名携带BTK点突变的患者外周血单个核细胞(PBMC)中的B细胞数量较少,但实际上发现其BTK转录本的量正常,因此我们推测该基因也会在其他谱系的细胞中表达。发现红白血病细胞系K-562、早幼粒细胞系HL-60和组织细胞淋巴瘤细胞系U-937的BTK mRNA水平与B细胞相当。在健康供体的单核细胞以及人未成熟嗜碱性细胞系KU812、人肥大细胞白血病细胞系HMC-1和表达CD34的成髓细胞KG-1中也检测到了BTK mRNA。通过免疫沉淀和Western印迹分析BTK蛋白时,获得了类似的表达模式。使用基于聚合酶链反应的分析方法,在T淋巴细胞中鉴定出少量(不到B细胞水平的1%)的BTK mRNA。我们的发现与BTK基因在造血细胞中的普遍表达一致,但在T淋巴细胞和浆细胞中,转录水平被选择性下调。