Lineage-specific expression of CYBB in leukocytes

(1)PU.1和HAF-1在CYB表达中的作用系统地诱导了CYBB启动子的单碱基替换，并检测了它们对PU.1和HAF-1的报告活性和结合亲和力的影响。数据表明，PU1是主要的，HAF-1是补充的。这也暗示了新的CGD，并为在CGD吞噬细胞中最大限度地表达该基因提供了理想的修饰启动子序列。(2)新的CW元件激活CyBB我们在CyBB启动子上发现了一个新的GATA结合位点。(3)IRF-4在B淋巴细胞中的作用IRF-4被认为是抑制CyBB的作用，这可能是导致GATA-1基因在B淋巴细胞中低表达的原因。(4)将GATA-1基因转染到具有异常的CYBB启动子的X-CGD患者的吞噬细胞中。为了使gp91Phox缺陷的中性粒细胞/单核细胞转染到gp91Phox正常的嗜酸性粒细胞，我们尝试将GATA-1基因转染到一名X连锁BP-CGT患者的外周血单核细胞中。脂质体和腺病毒转染法的效率较低，但非特异性地激活了该基因，提示该基因有可能被内源性激活。(5)7D5表位的鉴定。抗黄色素b5587D5表位的单抗从分子生物学角度定位于gp91Phox的胞外肽构象。该抗体现在无疑可用于X-CGD的诊断和监测CGD患者的基因转移效率。

(1) Roles of PU.1 and HAF-1 for the expression of CYBBSingle base substitutions were systematically induced to the CYBB promoter and examined their effects on reporter activities and binding affinities to PU.1 and HAF-1. Data suggested that PU.1 is principal and HAF-1 supplementary. These also implied new CGDs and supplied ideally modified promoter sequences for maximal expression of the gene in the CGD phagocytes.(2) A new cw-elemement for the activation of CYBBWe discovered a new GATA-binding site on the CYBB promoter. This site is essential for the expression of the gene promoted by PU.1 and GATA-1.(3) A role of IRF-4 in B lymphocytesIRF-4 was suggested to inhibit CYBB that may account for the low expression of gp91phox in B lymphocytes.(4) Trials for GATA-1 gene transfection to phagocytes of the X-CGD patient with an abnormal CYBB promoterFor making gp91phox-deficient neutrophils/monocytes to gp91phox-normal eosinophils, we tried to transfect GATA-1 gene to the peripheral blood monocytes of a patient with X-linked bp-53T CGD patient, Efficiencies of liposomal and adenoviral transfections were inefficient but activated the gene nonspecifically suggesting a potential of the gene to be activated endogenously.(5) Identification of the epitope for 7D5, a monoclonal antibody raised against flavocytochrome b5587D5 epitope was molecular-biologically located to the extracellular peptide conformation of gp91phox. The antibody is now undoubtedly available for diagnosis of X-CGD and monitoring the efficiency of the gene ttransfer to CGD patients.