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单链抗体scFv-M6-1B9对OKT3诱导的T细胞增殖具有强效抑制作用,并能抑制HeLa细胞中CD147细胞表面表达。

Potent inhibition of OKT3-induced T cell proliferation and suppression of CD147 cell surface expression in HeLa cells by scFv-M6-1B9.

作者信息

Intasai Nutjeera, Tragoolpua Khajornsak, Pingmuang Prakitnavin, Khunkaewla Panida, Moonsom Seangdeun, Kasinrerk Watchara, Lieber André, Tayapiwatana Chatchai

机构信息

Division ofClinicalMicroscopy,DepartmentofMedicalTechnology,FacultyofAssociatedMedicalSciences, Chiang MaiUniversity,ChiangMai50200,Thailand.

出版信息

Immunobiology. 2008;214(6):410-21. doi: 10.1016/j.imbio.2008.12.006.

Abstract

CD147, a multifunctional type I transmembrane glycoprotein, has been implicated in various physiological and pathological processes. It is involved in signal transduction pathways and also plays a crucial role in the invasive and metastatic activity of malignant tumor cells. Diminished expression of this molecule has been shown to be beneficial in suppression of tumor progression. In a previous study, we generated and characterized a recombinant antibody fragment, scFv, which reacted specifically to CD147. In the present study, we further investigated the biological properties, function and the effect of generated scFv on CD147 expression. The in vitro study showed that soluble scFv-M6-1B9 produced from E. coli HB2151 bound to CD147 surface molecule and inhibited OKT3-induced T cell proliferation. Furthermore, soluble lysate of scFv-M6-1B9 from 293A cells, transduced with a scFv-M6-1B9 expressing adenovirus vector, recognized both recombinant and native CD147. These results indicate that scFv-M6-1B9 binds with high efficiency and specificity. Importantly, scFv-M6-1B9 intrabody reduced the expression of CD147 on the cell surface of HeLa cells suggesting that scFv-M6-1B9 is biologically active. In conclusion, our present study demonstrated that scFv-M6-1B9 has a great potential to target both the intracellular and the extracellular CD147. The generated scFv-M6-1B9 may be an effective agent to clarify the cellular function of CD147 and may aid in efforts to develop a novel treatment in various human carcinomas.

摘要

CD147是一种多功能的I型跨膜糖蛋白,参与了多种生理和病理过程。它涉及信号转导途径,并且在恶性肿瘤细胞的侵袭和转移活动中也起着关键作用。已表明该分子表达的降低有利于抑制肿瘤进展。在先前的研究中,我们制备并鉴定了一种重组抗体片段scFv,它能特异性地与CD147反应。在本研究中,我们进一步研究了所产生的scFv的生物学特性、功能以及对CD147表达的影响。体外研究表明,由大肠杆菌HB2151产生的可溶性scFv-M6-1B9与CD147表面分子结合,并抑制OKT3诱导的T细胞增殖。此外,用表达scFv-M6-1B9的腺病毒载体转导的293A细胞中scFv-M6-1B9的可溶性裂解物能识别重组CD147和天然CD147。这些结果表明scFv-M6-1B9具有高效和特异性的结合能力。重要的是,scFv-M6-1B9胞内抗体降低了HeLa细胞表面CD147的表达,这表明scFv-M6-1B9具有生物学活性。总之,我们目前的研究表明,scFv-M6-1B9在靶向细胞内和细胞外CD147方面具有巨大潜力。所产生的scFv-M6-1B9可能是阐明CD147细胞功能的有效试剂,并可能有助于开发针对各种人类癌症的新疗法。

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