Transmission functions of fusion protein CTP-HBcAg18-27-Tapasin in the cells

Abstract

Abstract: AIM: To observe the function of transduction of fusion protein CTP-HBcAg18-27-Tapasin in antigen presenting cells.METHODS: Bone marrow derived DCs isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin-4 for 5 days followed by LPS added to induce DCs maturation. Then, different dosages of CTP-HBcAg18-27-Tapasin and RPMI-1640 were added to cell culture medium. Subsequently, the distribution and localization of intracellular immunofluorescence were observed by confocal microscopy with fluorescence intensity assaying quantitatively. Futhermore, the transduction efficiency of CTP-HBcAg18-27-Tapasin was evaluated by detecting the differences of expression of Tapasin using western blot in different group.RESULTS: DCs were cultured and identified successfully; Immunofluorescence confirmed the CTP-HBcAg18-27-Tapasin could penetrate the membrane into the cytoplasm of DCs but not into the nucleus. Fluorescence intensity of 50 μg/L CTP-HBcAg18-27-Tapasin group were higher than 10 μg/L CTP-HBcAg18-27-Tapasin group and blank groups. Western blot also confirmed that CTP-HBcAg18-27-Tapasin can penetrate the cell membrane into the DCs, and the data showed the difference was statistical.CONCLUSION: CTP-HBcAg18-27-Tapasin has the ability of penetrating and locating in the cytoplasm of DCs.

Key words: CTP, HBcAg18-27, Tapasin, Transduction, Dendritic cells

CLC Number:

R965.2

LIU Hong-hong, CHEN Xiao-hua, ZHOU Li-qin, LIU Xue-ni, YU Yong-sheng, ZANG Guo-qing, TANG Zheng-hao. Transmission functions of fusion protein CTP-HBcAg18-27-Tapasin in the cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(1): 25-29.

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