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...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。展开更多
Introduction:Butyrophilins(BTNs)belong to the immunoglobulin superfamily;they play crucial roles in immune regulation,especially inγδ T cell activation.While their expression has been studied in solid tumours,their ...Introduction:Butyrophilins(BTNs)belong to the immunoglobulin superfamily;they play crucial roles in immune regulation,especially inγδ T cell activation.While their expression has been studied in solid tumours,their involvement in hematologic malignancies remains poorly understood.Objectives:We hypothesised that BTNs are dysregulated in chronic lymphocytic leukaemia(CLL),contributing toγδT cell dysfunction and potentially influencing disease progression.Methods:In this study,we analyzed publicly available microarray and RNA-seq datasets to investigate the expression patterns of BTN genes in CLL.Results:Our findings reveal significant dysregulation of BTN gene expression in CLL,with BTN2A1,BTN3A1,BTN3A2,and BTN3A3 being markedly downregulated in peripheral blood mononuclear cells(PBMCs)and bone marrow samples from CLL patients compared to healthy volunteers,while BTN1A1 was upregulated.Furthermore,BTN2A2 was selectively downregulated in neoplastic B cells,whereas BTN3A1 was upregulated in T cells from CLL patients compared to healthy volunteers.Notably,lower BTN expression was associated with an unmutated IGVH status and male sex.Kaplan-Meier survival analysis demonstrated that higher expression of BTN2A1,BTN3A1,BTN3A2,and BTN3A3 correlated with a significantly longer overall survival.Conclusions:Given the established role of BTN2A1 and BTN3A1 in the phosphoantigen-mediated activation of Vδ2γδ T cells,their downregulation may contribute toγδ T cell dysfunction in CLL.These results highlight the potential prognostic value of BTN gene expression in CLL and underscore the need for further studies exploring its mechanistic role in disease progression and immune evasion.展开更多
文摘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。
基金funded by the National Science Center in Poland,Preludium grant number 2019/35/N/NZ6/02973.
文摘Introduction:Butyrophilins(BTNs)belong to the immunoglobulin superfamily;they play crucial roles in immune regulation,especially inγδ T cell activation.While their expression has been studied in solid tumours,their involvement in hematologic malignancies remains poorly understood.Objectives:We hypothesised that BTNs are dysregulated in chronic lymphocytic leukaemia(CLL),contributing toγδT cell dysfunction and potentially influencing disease progression.Methods:In this study,we analyzed publicly available microarray and RNA-seq datasets to investigate the expression patterns of BTN genes in CLL.Results:Our findings reveal significant dysregulation of BTN gene expression in CLL,with BTN2A1,BTN3A1,BTN3A2,and BTN3A3 being markedly downregulated in peripheral blood mononuclear cells(PBMCs)and bone marrow samples from CLL patients compared to healthy volunteers,while BTN1A1 was upregulated.Furthermore,BTN2A2 was selectively downregulated in neoplastic B cells,whereas BTN3A1 was upregulated in T cells from CLL patients compared to healthy volunteers.Notably,lower BTN expression was associated with an unmutated IGVH status and male sex.Kaplan-Meier survival analysis demonstrated that higher expression of BTN2A1,BTN3A1,BTN3A2,and BTN3A3 correlated with a significantly longer overall survival.Conclusions:Given the established role of BTN2A1 and BTN3A1 in the phosphoantigen-mediated activation of Vδ2γδ T cells,their downregulation may contribute toγδ T cell dysfunction in CLL.These results highlight the potential prognostic value of BTN gene expression in CLL and underscore the need for further studies exploring its mechanistic role in disease progression and immune evasion.
