Pajic Tadej, Cernelc Peter, Sesek Briski Alenka, Lejko-Zupanc Tatjana, Malesic Ivan
Department of Haematology, Division of Internal Medicine, University Medical Centre Ljubljana, Zaloska 7, SI-1000 Ljubljana, Slovenia.
Clin Biochem. 2009 Nov;42(16-17):1677-84. doi: 10.1016/j.clinbiochem.2009.08.003. Epub 2009 Aug 13.
To investigate the pattern of glutamate dehydrogenase (GLDH) activity, GLUD1 and GLUD2 expressions in peripheral blood mononuclear cells (PBMC) of untreated B-chronic lymphocytic leukemia (B-CLL) in healthy individuals (HI) and patients with infectious mononucleosis (IM).
GLDH activity was determined in a supernatant obtained from pelleted PBMC. GLUD1 and GLUD2 mRNA expression was determined using a quantitative real-time polymerase chain reaction. CD19(+) B cells from PBMC were purified by using positive selection.
The highest GLDH activity was found in PBMC of the B-CLL group followed by the HI group and IM group. The PBMC GLDH activity was higher in 60% of the B-CLL patients according to the established reference interval for our HI (2.17-5.70 microkat/g protein). The greater GLDH activity was also found in the CD19(+) cell preparation of the B-CLL patients (two of the three) but not in HI (n=3). The median value of GLUD1 expression was highest in the IM group (n=11), followed by the HI (n=14) and B-CLL groups (n=59) (median 4.69/3.78, P<0.005 and 4.69/2.91, P<0.0005, respectively). GLUD2 expression was not significantly different between groups.
The increased GLDH activity is specific for the PBMC of B-CLL patients. The GLUD1 but not the GLUD2 gene expression pattern is different between the PBMC of IM and B-CLL patients.
研究健康个体(HI)和传染性单核细胞增多症(IM)患者未经治疗的B细胞慢性淋巴细胞白血病(B-CLL)外周血单个核细胞(PBMC)中谷氨酸脱氢酶(GLDH)活性、GLUD1和GLUD2表达模式。
测定从沉淀的PBMC获得的上清液中的GLDH活性。使用定量实时聚合酶链反应测定GLUD1和GLUD2 mRNA表达。通过阳性选择从PBMC中纯化CD19(+) B细胞。
B-CLL组PBMC中GLDH活性最高,其次是HI组和IM组。根据我们HI组确定的参考区间(2.17 - 5.70微卡/克蛋白质),60%的B-CLL患者PBMC中GLDH活性更高。在B-CLL患者的CD19(+)细胞制剂中也发现了更高的GLDH活性(三例中的两例),但HI组(n = 3)中未发现。GLUD1表达的中位数在IM组(n = 11)中最高,其次是HI组(n = 14)和B-CLL组(n = 59)(中位数分别为4.69/3.78,P < 0.005和4.69/2.91,P < 0.0005)。各组间GLUD2表达无显著差异。
GLDH活性增加是B-CLL患者PBMC所特有的。IM和B-CLL患者的PBMC之间,GLUD1基因表达模式不同,而GLUD2基因表达模式无差异。