| OBJECTIVE:Human CD137L was demonstrated to be a type II glycoprotein. Human CD137/CD137L has been shown to amplify T-cell-mediated antitumor immunity in several mouse models.To detect the expression of serum mutational soluble CD137ligand (sCD137L) in acute myeloid leukemia (AML) patients and to explore the relationship between the expression and occurrence, development and prognosis of AML. CD137L was chosen as a costimulatory molecule in order to reinforce antitumor efficacy of human effector cells.METHODS:The sCD137L mutation was detected by reverse transcription polymerase chain reaction (RT-PCR) analysis in THP-1cells and60cases of AML with different primer. In this study, healthy person’s sCD137L cDNA amplification was used as positive control. The expression vector pETCD137L constructed by cloning the extracellular domain of CD137L into the expression vector pET30. The extracellular domain of CD137L was purified by affinity chromatography.4-1BBL expression was analyzed by SDS-PAGE and Western blot. The binding activity was examined by ELISA and immunohistochemistry.RESULTS:Two mutational sCD137Ls were detected in THP-1cells, named ml and m2mutation. The incidence of m1mutation was very low and relatively homogeneous. The incidence of m2mutation was decreasing in the following order, M2>M1=M6>M4>M5>M3. The m2mutation was closely related with AML disease progression. The data on DNA sequence confirmed that the CD137L protein was correctly constructed. The protein was recovered in high yield (up to12mg/L) and purity after His-tag purification.The purified soluble CD137L bound to A549and Jukate cells, respectivelyCONCLUSION:The change of sCD137L is related to the AML development and occurrence. The clinical testing of sCD137L is contributed to monitoring of the treatment and prognosis of AML. The extracellular domain of CD137LL has been successfully constructed and expressed in E.coli BL21(DE3), which could be useful in cancer immunotherapy. |
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