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Anti-tumor effects of murine T-lymphocytes harboring p185HER2 specific chimeric T-cell receptor gene
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Abstract:
Objective： To investigate the anti-tumor effects of murine T-lymphocytes harboring p185HER2 specific chimeric T-cell receptor gene.Methods: The p185HER2 specific chimeric T-cell receptor N29γ or N29ζ were introduced into retroviral vector pRET6, and the viral producer cell line was established using a Ping-Pong method. Murine spleen T-lymphocytes were transfected using an optimized protocol incorporating activation with immobilized anti-CD3/anti-CD28, followed by infection in the presence of Retronectin. Transduced murine T-lymphocytes were co-cultured with tumor cell lines overexpressing (SK-OV-3) or underexpressing (MCF-7) p185HER2 for assaying antigen specific cytokine release and CTL. Results: Endonuclease digestion showed the constructed vector was corrected. The titer of retroviral supernatant collected was 1.2×106 and the retroviral transduction efficiency reached over 50% with our optimized method. Both N29γ and N29ζ chimeric T-cell receptor transduced T-lymphocytes demonstrated HER2-specific antigen response as determined by release of interferon γ and cellular cytotoxicity assays. Conclusion: Our results suggest that murine T-lymphocytes harboring chimeric T-cell receptor gene had obvious antitumor effect in vitro through releasing cytokines and CTL effect.
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Project Supported:
NIH grant Ro1CA92488
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