Clinical significance of serum NGF level and CD271 expression in patients with acute B lymphoid leukemia / 中国实验血液学杂志

The aim of this study was to detect the nerve growth factor (NGF) level in serum and NGF low affinity acceptor CD271 expression on bone marrow leukemic cells in acute B lymphoid leukemia (B-ALL) patients and to analyze their clinical significance. The NGF level in serum and CD271 expression on leukemic cells in bone marrow were detected by enzyme linked immunosorbent assay and flow cytometry in B-ALL patients respectively. The results indicated that compared with control group, the NGF level in serum of patient group significantly increased (t = 4.191, p < 0.05), but CD271 expression on leukemic cells in bone marrow decreased significantly (t = 4.898, p < 0.05). The complete remission (CR) rate of 25 B-ALL patients was 64% (16/25) after one course of CVAD chemotherapy. There were statistically significant differences of NGF level and CD271 expression in non-remission (NR) group and control group (t = 3.976, p < 0.05 vs t = 5.052, p < 0.05), but there were no statistically difference of NGF level and CD271 expression in CR group (t = 1.102, p > 0.05 vs t = 1.150, p > 0.05) as compared with control group. The CD271 expression before and after chemotherapy between CR and NR groups showed statistically significant differences (t = 3.889, p < 0.05; t = 3.751, p < 0.05 and t = 4.678, p < 0.05 respectively), but NGF level before and after chemotherapy showed no statistical difference between these 2 groups (t = 0.476, p > 0.05). 50% (8/16) patients relapsed during following up, and of their NGF level [(168.00 ± 61.66) pg/ml] and CD271 expression [(52.29 ± 13.00)%] showed the significantly differences, compared with those in control group (t = 5.284, p < 0.05 vs. t = 6.073, p < 0.05), but the NGF level [(81.13 ± 25.32) pg/ml] and CD271 expression [(78.45 ± 7.12)%] of other 8 patients showed no statistical difference as compared with control group (t = 1.228, p > 0.05 vs t = 1.144, p > 0.05). Compared with low NGF level and CD271 low expression groups, the survival time of B-ALL patients with high NGF level and CD271 expression was not changed significantly (p = 0.750 vs p = 0.170). It is concluded that the increased NGF level in serum and decreased CD271 expression on bone marrow leukemic cells in B-ALL patients are related with leukemia development and may be the useful indexes to evaluate curative effect and prognosis.

Construction of lentiviral vector carrying mouse RORγt and expression of RORγt in 293FT cells / 中国实验血液学杂志

This study was aimed to construct a lentiviral vector carrying mouse RORγt and glp gene, and to detect the expression of RORγt in the 293FT cells. The RORγt fragment was amplified by RT-PCR from mouse thymus and cloned into PCR 2.1 vector. The RORγt DNA fragment was prepared by digestion and inserted into MigR1 plasmid, then the RORγt-IRES-GFP was directionally linked with lentiviral transfer plasmid pTK208 to generate a lentiviral vector pXZ9-RORγt. The recombinant lentivirus were produced by co-transfected three plasmids into 293FT packing cells using lipofectamine 2000. After transfection, the lentiviral supernatant was collected and concentrated via ultracentrifugation. The 293FT cells were infected by the concentrated lentivirus, GFP expression was examined under a fluorescent microscope and the expression of RORγt protein was detected by Western blot. The results showed that the RORγt fragment was amplified from cDNA of mouse thymus and recombinant lentiviral vector pXZ9-RORγt was constructed successfully. High titer lentivirus were prepared after one round ultracentrifugation. RORγt expression could be detected in 293FT cells after virus infection. It is concluded that the lentiviral vector pXZ9- RORγt containing mouse RORγt-IRES-GFP is successfully constructed; RORγt can express in 293FT cells via lentiviral vector transduction, which provides an optional tool for further research on the mechanism of RORγt controlling Th17 cell differentiation.