The levels of CD3[epsilon], LCK, VAV1, and ZAP70 gene expression were evaluated by quantitative real-time PCR (Fig.
In order to determine the activation of T cell receptor signaling pathway, we studied the correlation among CD3[epsilon], LCK, VAV1, and ZAP70 genes in HTLV-1-infected individuals.
Therefore, we performed the correlation between FOXP3 gene expression and the CD3[epsilon], LCK, VAV1, and ZAP70 genes in HTLV-1-infected individual.
CD3[epsilon], LCK, VAV1 are correlated to PVL and Tax expression Since CD3[epsilon], LCK, VAV1, and ZAP70 genes were upregulated in HAM/TSP compared to HAC group; we suggested that these genes could be correlated to the PVL and Tax expression.
More recently, in 2010, investigators proposed a molecular-scoring system based on a combination of tests for the expression of the ZAP70, LPL (lipoprotein lipase), CLLU1 (chronic lymphocytic leukemia upregulated 1), MIR29C (microRNA 29c), and MIR223 (microRNA 223) genes that seemed to predict patient outcomes (6).
4] Human genes: TP53, tumor protein p53; ZAP70, zeta-chain (TCR) associated protein kinase 70kDa; LPL, lipoprotein lipase; CLLU1, chronic lymphocytic leukemia up-regulated 1; MIR29C, microRNA 29c; MIR223, microRNA 223.
We describe the validation of a new standardized quantitative real-time reverse transcription-PCR (qPCR) analysis for measuring ZAP70 mRNA in purified B lymphocytes and its power as both a surrogate for IGHV mutational status and a prognostic marker for survival and treatment-free time in CLL.
We measured the expression of cytoplasmic ZAP70 protein by FC with the Fix and Perm Permeabilization Kit (ImTec Diagnostics), a ZAP70 phycoerythrin-conjugated antibody (clone 1E7.
A calibrator sample (cDNA from the Namalwa cell line, a human B-lymphoid leukemia cell line that expresses ZAP70 at a low level; ATCC) was included as a control in each experiment.
We analyzed intracellular ZAP70 expression with flow cytometry according to Crespo et al.
We analyzed ZAP70 and LPL expression by measuring mRNA with qPCR on an ABI Prism 7700 Sequence Detector (Applied Biosystems) or a Bio-Rad IQ5 Real Time Detection System (Bio-Rad).
ROC curve analyses were performed with MedCalc statistical software (MedCalc Software) to determine the ZAP70 and LPL expression cutoff values that best distinguished between mutated and unmutated cases.