Human umbilical cord-drived mesenchymal stem cells as vehicles of CD20 specific-TRAIL fusion protein against non-Hodgkin’ s lymphoma / 中国免疫学杂志

ABSTRACT

Objective:To study the therapeutic effect of a novel double-target system,in which human umbilical cord-derived MSCs were used as vehicles to deliver fusion protein scFvCD20sTRAIL to non-Hodgkin ’ s lymphoma. Methods: The traditional methods in molecular biology were used to construct lentivirus expression vectors pLenR. scFvCD20 sTRAIL and contrast vectors. Human umbilical cord-derived MSCs ( HUMSCs ) were labeled with the copGFP by transducing with pseudo viral particles which had been packaged in 293T cells with four plasmid-lentivirus packaging system. Fusion protein scFvCD20sTRAIL were secreted from MSC. scFvCD20sTRAIL after that HUMSCs were infected by pseudo viral particles. CCK8 assay was applied to detect the antigen-restricted cell death induced by scFvCD20sTRAIL in CD20-positive BJAB and Raji cells as well as CD20-negtive Jurkat cells and human normal peripheral blood mononuclear cells (PBMCs). To evaluate the therapeutic effect of MSC. scFvCD20sTRAIL in vivo,ge-netically modified HUMSCs were intravenously injected into tumor-bearing mice with BJAB cells. The volume of tumor was measured every three days, and the inhibition ratio of tumor was calculated according to tumor volume. Results: Lentivirus expression vectors pLenR. scFvCD20sTRAIL, pLenR. ISZsTRAIL, pLenR. scFvCD20 and pLenR. CopGFP were successfully constructed and these constructs could be expressed stably in HUMSCs by lentivirus transduction. scFvCD20sTRAIL fusion protein produced a potent inhibition of cell proliferation in CD20-positive BJAB cells,moderate inhibition of the growth of Raji cells,and weak inhibition in CD20-negtive Jurkat cells when compared with ISZ-sTRAIL treatment,and it had no effect on normal human peripheral blood mononuclear cells (PBMCs). The MSC. scFvCD20sTRAIL treatment significantly inhibited the tumor growth when compared with those treated with MSC. ISZ-sTRAIL. Conclusion: A double-target therapeutic system is well established, in which HUMSCs migrated to tumor site, secreted a novel fusion protein scFvCD20sTRAIL,and thus locally concentrated scFvCD20sTRAIL extended antigen-restricted anti-tumor activity. The engineered HUMSCs secreting scFvCD20sTRAIL showed potent effect on inhibiting tumor growth in BJAB lymphoma malignancy,which may play an essential role in the clinical research .