| 350 | 0 | 43 |
| 下载次数 | 被引频次 | 阅读次数 |
本研究旨在通过生物信息学方法，结合对人体的肿瘤组织切片进行免疫组织化学检测，分析免疫调节分子BTN2A2 (butyrophilin subfamily 2 member A2)结构及其在肿瘤组织中的表达，为研究BTN2A2分子的免疫调节功能提供理论和实验支撑。序列分析表明，BTN2A2具有IgV样结构域，与B7家族分子具有序列同源性和结构相似性。系统发育树分析表明，BTN2A2和已知B7家族分子之间具有亲缘性。通过I-TASSER成功构建BTN2A2蛋白三维晶体结构模型，hBTN2A2配体为Cys,配体结合位点相应氨基酸残基分别为Arg-54、His-56、Ser-111、Val-112和Ala-113。蛋白质-蛋白质相互作用网络(protein-protein interaction, PPI)方法表明，存在10个与BTN2A2相关的分子，其中CD4和CD8与T细胞免疫相关。ZDOCK分析表明，BTN2A2确实可以与T细胞表面CD4和CD8分子结合。ClusterProfiler分析表明，BTN2A2和相关分子主要集中在原发性免疫缺陷、抗原加工和呈递、T细胞受体信号通路等免疫调节方面。流式细胞术(flow cytometry, FCM)分析进一步表明，BTN2A2蛋白与T细胞表面CD4和CD8分子间存在相互作用，且BTN2A2蛋白可抑制CD4+和CD8~+T细胞增殖和活化(P<0.001)。免疫组织化学染色显示，hBTN2A2在人肺癌、肝癌、肾癌和食管癌组织中的表达高于正常组织，而在卵巢癌组织未见表达(P<0.05);PrognoScan数据库和Kaplan-Meier曲线进行肺癌患者生存分析表明，hBTN2A2高表达的患者预后良好(P<0.001);TIMER数据库分析表明，肺癌中hBTN2A2表达水平与免疫细胞浸润呈正相关关系；GEPIA数据库分析显示，肺癌中hBTN2A2表达主要与CD8+ T细胞、CD3+ T细胞、树突状细胞(dendritic cell, DC)、辅助性T细胞(helper T cell, Th)1等免疫细胞浸润有关(P<0.001)。研究结果为免疫调节分子BTN2A2结构和功能性表达的深入研究提供了理论基础。
Abstract：The study aims to analyze the structure of the immunomodulatory molecule BTN2 A2 and its expre-ssion in different tumor tissues through bioinformatics methods, immunohistochemistry techniques and human tumor tissue sections, which provides theoretical and experimental support for studying the immunomodulatory function of BTN2 A2 molecules. Sequence analysis showed that BTN2 A2 had an IgV-like domain, and had sequence homology and structural similarity with B7 family molecules. Phylogenetic tree analysis showed that BTN2 A2 and the species of known B7 family molecules had affinity. The three-dimensional crystal structure model of BTN2 A2 protein was successfully constructed by I-TASSER. The hBTN2 A2 ligand was Cys, and the corresponding amino acid residues of the ligand binding site were Arg-54, His-56, Ser-111, Val-112 and Ala-113, respectively. The protein-protein interaction(PPI) network screened out 10 molecules related to BTN2 A2, among which the CD4 and CD8 were related to T cell immunity. ZDOCK molecular docking analysis showed that BTN2 A2 could indeed bind to CD4 and CD8 molecules on the surface of T cells. ClusterProfiler analysis showed that BTN2 A2 and related molecules mainly focused on primary immunodeficiency, antigen processing and presentation, T cell receptor signaling pathway and other immune regulation. Flow cytometry(FCM) analysis further showed that there was an interaction between BTN2 A2 protein and CD4 and CD8 molecules on the surface of T cells, and BTN2 A2 protein could inhibit the proliferation and activation of CD4+ and CD8+ T cells(P<0.001). Immunohistochemical staining showed that the expression of hBTN2 A2 in human lung, liver, kidney, and esophageal cancer tissues was higher than that in normal tissues, but no expression in ovarian cancer tissues(P<0.05); PrognoScan database and Kaplan-Meier curve for lung cancer patient survival analysis showed that lung cancer prognosis of patients with high expression of hBTN2 A2 in lung cancer was good; TIMER and database analysis showed that the expression of hBTN2 A2 in lung cancer was positively correlated with immune cell infiltration; GEPIA database analysis showed that hBTN2 A2 expression in lung cancer was mainly related to the infiltration of CD8+ T, CD3+ T cells, dendritic cells(DC), helper T cells(Th) 1 and other immune cells(P<0.001). The research results provide a theoretical basis for the in-depth study of the structure and functional expression of the immunomodulatory molecule BTN2 A2.
Ammann J.U.,Cooke A.,and Trowsdale J.,2013,Butyrophilin Btn2a2 inhibits TCR activation and phosphatidylinositol 3-kinase/Akt pathway signaling and induces Foxp3 expression in T lymphocytes,J.Immunol.,190(10):5030-5036.
Arnett H.A.,and Viney J.L.,2014,Immune modulation by butyrophilins,Nat.Rev.Immunol.,14(8):559-569.
Barber D.L.,Wherry E.J.,Masopust D.,Zhu B.G.,Allison J.P.,Sharpe A.H.,Freeman G.J.,and Ahmed R.,2006,Restoring function in exhausted CD8 T cells during chronic viral infection,Nature,439(7077):682-687.
Borst J.,Ahrends T.,Baba?a N.,Melief C.J.M.,and Kastenmüller W.,2018,CD4+ T cell help in cancer immunology and immunotherapy,Nat.Rev.Immunol.,18(10):635-647.
Bullock T.N.,and Yagita H.,2005,Induction of CD70 on dendritic cells through CD40 or TLR stimulation contributes to the development of CD8+ T cell responses in the absence of CD4+ T cells.J.Immunol.,174(2):710-717.
Doering T.A.,Crawford A.,Angelosanto J.M.,Paley M.A.,Ziegler C.G.,and Wherry E.J.,2012,Network analysis reveals centrally connected genes and pathways involved in CD8+ T cell exhaustion versus memory,Immunity,37(6):1130-1144.
Freeman G.J.,Gray G.S.,Gimmi C.D.,Lombard D.B.,Zhou L.J.,White M.,Fingeroth J.D.,Gribben J.G.,and Nadler L.M.,1991,Structure,expression,and T cell costimulatory activity of the murine homologue of the human B lymphocyte activation antigen B7,J.Exp.Med.,174(3):625-631.
Gavitt A.W.,Kirk D.J.,and David M.J.,2016,Lung cancer staging and prognosis,Cancer Treatment and Research,170:47-75.
Ghosh S.,Sarkar M.,Ghosh T.,Guha I.,Bhuniya A.,Biswas J.,Mallick A.,Bose A.,and Baral R.,2016,Absence of CD4(+) T cell help generates corrupt CD8(+) effector T cells in sarcoma-bearing Swiss mice treated with NLGP vaccine,Immunol.Lett.,175:31-39.
Huang W.S.,and August A.,2015,The signaling symphony:T cell receptor tunes cytokine-mediated T cell di-fferentiation,J.Leukoc.Biol.,97(3):477-485.
Kakimi K.,Karasaki T.,Matsushita H.,and Sugie T.,2017,Advances in personalized cancer immunotherapy,Breast Cancer,24(1):16-24.
Kythreotou A.,Siddique A.,Mauri F.A.,Bower M.,and Pinato D.J.,2018,Pd-L1,J.Clin.Pathol.,71(3):189-194.
Liu H.Z.,Wang Y.S.,Zeng Q.H.,Zeng Y.Q.,Liang C.L.,Qiu F.F.,Nie H.,and Dai Z.H.,2017,Suppression of allograft rejection by CD8+CD122+PD-1+ tregs is dictated by their Fas ligand-initiated killing of effector T cells versus Fas-mediated own apoptosis,Oncotarget,8(15):24187-24195.
Ma W.S.,Liang J.M.,Mo J.J.,Zhang S.Y.,Hu N.D.,Tian D.B.,and Chen Z.S.,2021,Butyrophilin-like 9 expression is associated with outcome in lung adenocarcinoma,BMC Cancer,21(1):1096.
Munn D.H.,2018,The host protecting the tumor from the host-targeting PD-L1 expressed by host cells,J.Clin.Invest.,128(2):570-572.
Ni L.,and Dong C.,2017,New B7 family checkpoints in human cancers,Mol.Cancer Ther.,16(7):1203-1211.
Palucka K.,and Banchereau J.,2012,Cancer immunotherapy via dendritic cells,Nat.Rev.Cancer,12:265-277.
Rhodes D.A.,Reith W.,and Trowsdale J.,2016,Regulation of immunity by butyrophilins,Annu.Rev.Immunol.,34:151-172.
Robainas M.,Otano R.,Bueno S.,and Ait-Oudhia S.,2017,Understanding the role of PD-L1/PD1 pathway blockade and autophagy in cancer therapy,Onco.Targets Ther.,10:1803-1807.
Sarkar B.,Kulharia M.,and Mantha A.K.,2017,Understanding human thiol dioxygenase enzymes:structure to function,and biology to pathology,Int.J.Exp.Pathol.,98(2):52-66.
Smith I.A.,Knezevic B.R.,Ammann J.U.,Rhodes D.A.,Aw D.,Palmer D.B.,Mather I.H.,and Trowsdale J.,2010,BTN1A1,the mammary gland butyrophilin,and BTN2A2 are both inhibitors of T cell activation,J.Immunol.,184(7):3514-3525.
Topalian S.L.,Taube J.M.,Anders R.A.,and Pardoll D.M.,2016,Mechanism-driven biomarkers to guide immune checkpoint blockade in cancer therapy,Nat.Rev.Cancer,16(5):275-287.
Wherry E.J.,and Kurachi M.,2015,Molecular and cellular insights into T cell exhaustion,Nat.Rev.Immunol.,15(8):486-499.
Zhang M.,Wang C.L.,and Cao Y.D.,2019,The relationship between tumor-infiltrating lymphocytes and the treatment and prognosis of breast cancer,Genomics and Applied Biology,38(11):5357-5362.(张铭，王成龙，曹友德，2019,肿瘤浸润淋巴细胞与乳腺癌治疗及预后的关系，基因组学与应用生物学，38(11):5357-5362.)
基本信息:
DOI：10.13417/j.gab.041.001550
中图分类号:R734.2
引用信息:
[1]杨文江,何雪萍,黄佑娇,等.免疫调节分子 BTN2A2结构及其在肺癌表达的分析[J].基因组学与应用生物学,2022,41(07):1550-1566.DOI:10.13417/j.gab.041.001550.
基金信息:
国家自然科学基金资助项目(NSFC 81660033); 贵州省普通高校科技拔尖人才支持计划项目(黔教合KY字[2017]071); 贵州省科技计划项目(黔科合支撑[2020]4Y230号); 贵州省国际科技合作基地项目(黔科合平台人才[2020]4103号)共同资助
2022-03-18
2022-03-18
2022-03-18