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CD80-streptavidin-decorated CNE2 cells enhance cytotoxicity of T cells
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Abstract:
Objective:To prepare CD80-streptavidin (CD80-SA) fusion protein and immobilize it on the surface of nasopharyngeal carcinoma CNE2 cells (CD80-SA-CNE2 cells), so as to investigate the effect of CD80-SA-CNE2 cells on cytotoxicity of T cells. Methods: pET21a-CD80-SA-6His expression plasmid was transformed into E. coli BL21 (DE3). The CD80-SA fusion protein was induced by IPTG, purified by Ni-NTA affinity chromatography and refolded by dialysis. The immobilization rate of CD80-SA on CNE2 cell surface was analyzed by flow cytometry, and the effect of CD80-SA-CNE2 cells on cytotoxicity of T cells was detected by LDH assay. Results: CD80-SA fusion protein was successfully prepared and purified. CD80 was lowly expressed on CNE2 cells (\[2.233±0.176\]%). CD80-SA fusion protein was effectively immobilized on the surface of biotinylated-CNE2 cells, with the immobilization rate being 73%. Moreover, CD80-SA-CNE2 cells effectively induced cytotoxicity of T cells; the cytotoxic rates of T cells were (37±3.12)%, (51±263)% and (58±2.47)% at the E∶T ratios of 1∶1, 1∶20 and 1∶40, respectively, which were significantly higher than those of control CNE2 cells (all P<0.01). Conclusion: CD80-SA fusion protein can be effectively immobilized on the surface of biotinylated-CNE2 cells, enhancing the cyctotoxicty of T cells against CNE2 cells.
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Project Supported:
Project supported by the National High Technology Research and Development Program (863 Program) of China（No. 2006AA02Z4C4）, and the Science and Technology Research Program of Baiyun District of Guangzhou City (No. 2009-SZ-40)
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