免疫原性HER2突变体蛋白对小鼠T细胞亚群分化的影响

宁宏宇; 江涛; 张瑞; 何羽; 姚文兵; 田浤

doi:10.11665/j.issn.1000-5048.20180115

免疫原性HER2突变体蛋白对小鼠T细胞亚群分化的影响

宁宏宇^1,2,
江涛^1,2,
张瑞^1,2,
何羽^1,2,
姚文兵^1,2,
田浤^1,2

1.
中国药科大学江苏省生物药物成药性研究重点实验室
2.
中国药科大学生命科学与技术学院

基金项目: 国家自然科学基金资助项目(No.81273426,No.81673343)

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- 刊出日期: 2018-02-24

Effects of immunogenic HER2 on the differentiation of T-cell in mouse

NING Hongyu^1,2,
JIANG Tao^1,2,
ZHANG Rui^1,2,
HE Yu^1,2,
YAO Wenbing^1,2,
TIAN Hong^1,2

摘要

摘要: 为了考察蛋白质中定点引入对硝基苯丙氨酸后对T细胞分化的影响，以人表皮生长因子受体2(HER2)胞外区为模型分子，利用实验室前期构建的工程菌表达了定点引入对硝基苯丙氨酸的HER2突变体。用含对硝基苯丙氨酸的HER2突变体免疫小鼠，分析其对小鼠T细胞亚群分化的影响。研究结果显示，对硝基苯丙氨酸的引入显著增强了HER2的免疫原性，免疫5周后，突变体免疫组的小鼠体内抗HER2抗体滴度明显高于PBS组和原型免疫组的小鼠；流式细胞术检测结果显示，小鼠淋巴结内Tfh细胞比例从4%上升至8%，而Treg细胞比例从5%下降至1%；QT-PCR分析结果显示，Tfh细胞关键转录因子Bcl-6的mRNA水平提高了2倍，而Treg细胞关键转录因子FOXP3的mRNA水平明显降低；ELISA法检测血清中Tfh细胞效应因子IL-21的含量从4 pg/mL上升至28 pg/mL。上述研究结果证实，含有对硝基苯丙氨酸的HER2突变体可以通过激活Tfh细胞分化而引起高效免疫应答。
- 免疫原性氨基酸 /
- HER2 /
- 滤泡辅助性T细胞 /
- 调节型T细胞 /
- 细胞分化
Abstract: In order to investigate the effects of p-nitrophenylalanine on the differentiation of T lymphocyte, human epidermal growth factor receptor 2(HER2)mutants were constructed by incorporating p-nitrophenylalanine to the extracellular domain of HER2. Mice were immunized with HER2 mutant to analyze the effects of p-nitrophenylalanine on the differentiation of T cell subsets. The results showed that the immunogenicity of HER2 protein was significantly enhanced by incorporation of immunogenic amino acids. HER2 mutants induced production of more anti-WT-HER2 antibody and greater titer of antibody than PBS and wild-type HER2 in the fifth week. The differentiation frequency of Th1 and Th2 cells showed no significant difference, but the differentiation frequency of Tfh cell was found to increase from 4% to 8%, while the differentiation frequency of Treg cell was found to decrease from 5% to 1% through flowcytometry analysis. The gene expression of Bcl-6 and FOXP3 were determined by quantitative real-time PCR. The results showed that the expression of Bcl-6 mRNA increased about 2 fold, while the expression of FOXP3 mRNA could be decreased to about 50% in HER2 mutant-immunized group. The secretion level of IL-21 in serum was also detected to increase from 4 to 28 pg/mL in HER2 mutant-immunized group. The above results confirm that HER2 mutants containing p-nitrophenylalanine can cause an efficient immune response by activating Tfh cell differentiation.
- immunogenic amino acid /
- HER2 /
- follicular helper T cell /
- regulatory T cell /
- cellular dfferentiation

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参考文献(20)

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免疫原性HER2突变体蛋白对小鼠T细胞亚群分化的影响

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出版历程

Effects of immunogenic HER2 on the differentiation of T-cell in mouse

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出版历程

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