LI Jing, YE Zheng-xu, YANG Shou-jing, LI Kai-nan, GAO Juan and LIU Yan-fang. Hsp70-NP Recombinant Protein Enhances HTNV NP Specific Cellular Immune Response in vivo[J]. Virologica Sinica, 2006, 21(4): 339-342.
Citation:
LI Jing, YE Zheng-xu, YANG Shou-jing, LI Kai-nan, GAO Juan, LIU Yan-fang.
Hsp70-NP Recombinant Protein Enhances HTNV NP Specific Cellular Immune Response in vivo .VIROLOGICA SINICA, 2006, 21(4)
: 339-342.
Hsp70-NP融合蛋白增强诱导HTNV NP特异性CTL的实验研究
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李静
1
,
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叶正旭
2
,
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杨守京
1*,,
,
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李铠男
1
,
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高娟
3
,
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刘彦仿
1
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摘要
本文采用纯化蛋白Hsp70-NP,NP,Hsp70分别免疫C57/BL6小鼠,取各组小鼠脾淋巴细胞进行淋巴细胞增殖试验和细胞毒试验。此外,为了获得细胞毒实验的靶细胞,本文还采用脂质体介导质粒pcDNA3.1/S转染黑色素瘤细胞B16,通过G418筛选稳定克隆,并用RT-PCR,Western blots 以及免疫荧光染色证实N蛋白在胞浆中表达。淋巴细胞增殖实验表明,Hsp70-NP,NP组小鼠脾淋巴细胞均能够对体外抗原刺激产生增殖反应,而Hsp70-NP组的增殖指数明显高于NP免疫组。细胞毒实验结果表明,LDH的释放具有效应细胞依赖性,Hsp70-NP,NP免疫组脾淋巴细胞均可以特异性杀伤靶细胞B16-N,而Hsp70-NP免疫组的杀伤率显著高于NP免疫组。实验结果显示,Hsp70可以增强NP诱导产生特异性CTL的能力。本研究结果为进一步设计基于NP的合成肽疫苗或基因疫苗提供了重要实验依据。
Hsp70-NP Recombinant Protein Enhances HTNV NP Specific Cellular Immune Response in vivo
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1.
1.Department of Pathology
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2.
Department of Orthopaedics
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3.
Department of Gastroenterology, Xi Jing hospital, the Fourth Military Medical University, Xi’an 710032, China
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Corresponding author:
YANG Shou-jing,
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Abstract
To examine whether the recombinant protein Hsp70-NP could induce the N-specific CTLs, we immunized C57/BL6 mice with purified protein Hsp70-NP, NP or Hsp70. The lymphoproliferative responses and CTL activities of spleen cells from vaccinated mice were determined. Accordingly, in order to get the target cells for cytotoxic assay, we tranfected pcDNA3.1/S into mice melanoma cells B16 with Lipofectamine2000, and selected stable transfectants with G418. RT-PCR, Western blots and immunofluorescence were used to identify the B16-N. The lymphoproliferative responses of spleen cells of mice vaccinated with Hsp70-NP, NP clearly showed proliferation in the presence of the N protein. But the stimulation indexes of splenocytes to NP of Hsp70-NP group were significantly higher than that of NP group. The CTL results showed that the cytotoxic effect of lymphocyte of the Hsp70-NP group was higher than that of the NP gorup. The results showed that Hsp70 could enhance the potency of NP to elicit HTNV NP specific cellular immune responses in vivo.
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References
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Proportional views
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