免疫原性HER2突变体蛋白对小鼠T细胞亚群分化的影响
Effects of immunogenic HER2 on the differentiation of T-cell in mouse
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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。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.
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[1] Gruenewald J,Tsao ML,Perera R,et al.Immunochemical termination of self-tolerance[J].Proc Natl Acad Sci U S A,2008,105(32):11276-11280. [2] Hardy LL, Wick DA, Webb JR. Conversion of tyrosine to the inflammation-associated analog 3′-nitrotyrosine at either TCR-or MHC-contact positions can profoundly affect recognition of the MHC class I-restricted epitope of lymphocytic choriomeningitis virus glycoprotein 33 by CD8 T cells[J].J Immunol,2008,180(9):5956-5962. [3] Xie J,Schultz PG.An expanding genetic code[J].Methods,2005,36(3):227-238. [4] Kessel C,Nandakumar KS,Peters FB,et al.A single functional group substitution in C5a breaks B cell and T cell tolerance and protects against experimental arthritis[J].Arthritis Rheumatol,2014,66(3):610-621. [5] Hallam TJ, Wold E, Wahl A, et al. Antibody conjugates with unnatural amino acids[J].Mol Pharm,2015,12(6):1848-1862. [6] Grunewald J,Hunt GS,Dong L,et al.Mechanistic studies of the immunochemical termination of self-tolerance with unnatural amino acids[J].Proc Natl Acad Sci U S A,2009,106(11):4337-4342. [7] Gauba V, Grunewald J, Gorney V, et al. Loss of CD4 T-cell-dependent tolerance to proteins with modified amino acids[J].Proc Natl Acad Sci U S A,2011,108(31):12821-12826. [8] Zhang TJ,Tian H,Gao XD.Advances in research on antitumor activities of HER2-based peptide vaccines[J].Prog Pharm Sci(药学进展),2013,37(10):516-521. [9] Zhang R,Tian H,Gao XD,et al.Application of next generation genome editing technology in gene therapy and biopharmaceuticals[J].J China Pharm Univ(中国药科大学学报),2014,45(4):504-510. [10] Yao DN,Xu XW,Yu HB,et al.Screening system for amber suppressor tRNA based on inducible high-copynumber plasmid[J].J China Pharm Univ(中国药科大学学报),2013,44(2):174-178. [11] Chen X,Ma W,Zhang T,et al.Phenotypic Tfh development promoted by CXCR5-controlled re-localization and IL-6 from radiation-resistant cells[J].Protein Cell,2015,6(11):825-832. [12] Moser B.CXCR5,the defining marker for follicular B helper T(T-FH)cells[J].Front Immunol,2015,6:296. [13] Mcguire HM,Vogelzang A,Warren J,et al.IL-21 and IL-4 collaborate to shape T-dependent antibody responses[J].J Immunol,2015,195(11):5123-5135. [14] Choi YS,Yang JA,Yusuf I,et al.Bcl6 expressing follicular helper CD4 T cells are fate committed early and have the capacity to form memory[J].J Immunol,2013,190(8):4014-4026. [15] Liu X,Yan X,Zhong B,et al.Bcl6 expression specifies the T follicular helper cell program in vivo[J].J Exp Med,2012,209(10):1841-1852. [16] Ozaki K,Spolski R,Ettinger R,et al.Regulation of B cell differentiation and plasma cell generation by IL-21,a novel inducer of Blimp-1 and Bcl-61[J].J Immunol,2004,173(9):5361-5371. [17] Crotty S. A brief history of T cell help to B cells[J].Nat Rev Immunol,2015,15(3):185-189. [18] Toellner KM.Cognate interactions:extrafollicular IL-4 drives germinal-center reactions,a new role for an old cytokine[J].Eur J Immunol,2014,44(7):1917-1920. [19] Zhang Y,Garcia-Ibanez L,Toellner KM.Regulation of germinal center B-cell differentiation[J].Immunol Rev,2016,270(1):8-19. [20] Dan JM,Arlehamn CSL,Weiskopf D,et al.A cytokine-independent approach to identify antigen-specific human germinal center T follicular helper cells and rare antigen-specific CD4(+)T cells in blood[J].J Immunol,2016,197(3):983-993. -
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