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cwq921: ½ð±Ò+50, ¡ï¡ï¡ï¡ï¡ï×î¼Ñ´ð°¸, °ïʵÑéÊҵģ¬Ð»Ð»À²£¡ 2014-12-15 12:45:43
cwq921: ½ð±Ò+50, ¡ï¡ï¡ï¡ï¡ï×î¼Ñ´ð°¸, °ïʵÑéÊҵģ¬Ð»Ð»À²£¡ 2014-12-15 12:45:43
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Abstract: To investigate the effect of expression of SIRP¦Á-Fc (SF), a fusion protein encoded by adenovirus Ad35-SF, on T cell function, Jurkat cells were infected with adenovirus Ad35-SF for 48 h. Western blotting analysis showed that the expressed fusion protein SF was about 48 kD, which was consistent with the calculated value. ELISA assay showed that the concentration of the fusion protein in the culture medium was 19.1 ¦Ìg/ml. CD47-positive cells decreased dramatically from 97.06% to 15.32% when Jurkat cells were infected with Ad35-SF [multiplicity of infection (MOI)=1] for 8h, and further decreased to 2.94% and 1.73%, respectively, if MOI was increased to 5 and 10. The proliferation of Jurkat cells 48 h after Ad35-SF infection increased by 29% and increased the killing effect of of the infected Jrukat cells to hepatocellular carcinoma cell line Hep3B by 25.6%, as compared with the controls. The results of this study showed that the adenovirus Ad35-SF we made can efficiently infect Jurkat cells and express functional secretory protein SF, thereby blocking the cell surface marker CD47. In addition, the infected Jurkat cells will continue to secrete SF protein and block other uninfected Jurkat cells, which can significantly improve the proliferation of Jurkat cells and their cytotoxicity to hepatocellular carcinoma cells. ×¢£º 1¡¢Ã»ÓÐÍêÈ«°´ÕÕÄãµÄÖÐÎÄÒ»¶ÔÒ»·Ò루¾¡¿ÉÄܰ´ÄãµÄÔ»°£©£¬¶øÊǸù¾ÝÓ¢Îıí´ïϰ¹ß°´ÕÕÄãÒª±í´ïµÄÄÚÈÝ×öÁËÊʵ±µ÷Õû¡£ 2¡¢ÄãµÄÖÐÎÄÓÐÐí¶àµØ·½ÐèÒªµ÷Õû£¬ÈçµÚÒ»¾äºÍµÚ¶þ¾äÖ®¼äÓÐÒ»ÖÖÄ¿µÄ¹ØÏµ£¬¹Ê¶ø²»ÒËÓþäºÅ¡£µ¹ÊǺóÃæÒ»Á¬´®µÄ¶ººÅÐèÒªÖØÐ¶Ͼ䡣ÔÙÈ硱ϸ°ûÉÏÇåÖеıí´ïÁ¿Îª19.1¦Ìg/ml¡°£¬ÎÒÃ÷°×ÄãÔÚ˵ʲô£¬µ«Ñϸñ˵À´Ëùνϸ°ûÉÏÇåÓ¦¸ÃÊÇÖ¸ÀëÐĺóµÄÅàÑø»ù£¨Ïà¶ÔÓÚÌõ¼þÅàÑø»ùconditional medium£©£¬¹Ê¶øÎÒÔÚ·ÒëʱÓÃÁËculture medium¶ø·Çcell supernatants£»ÁíÍ⣬ÄãÈ·¶¨µ°°×Ũ¶ÈÊÇ19.1 ¦Ìg/ml¶ø²»ÊÇ19.1 ng/mlÂ𣿠ϣÍûÓаïÖú£¬¾¡Ôç±ÏÒµ£¡ |
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