T Cell Antigen Recognition

T细胞抗原识别

• 批准号: 7929506
• 负责人: ELLIS L REINHERZ
• 金额: $ 43.62万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-09 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7929506
• 关键词: Accounting; Affect; Affinity; Antibodies; Antigen-Presenting Cells; Antigens; Autoimmune Process; Award; Binding; Binding Sites; C-terminal; CD3 Antigens; Cell surface; Complex; Development; Electron Spin Resonance Spectroscopy; Elements; Environment; Escherichia coli; Event; Fetal Thymic Organ Culture; Funding; Grant; Histocompatibility Antigens Class II; Human; Immunologic Deficiency Syndromes; Ligands; Ligation; Liposomes; MHC Class I Genes; MHC Class II Genes; MHC Interaction; Measurement; Measures; Mediating; Membrane; Methods; Modeling; Molecular; Molecular Conformation; Monoclonal Antibodies; Mutation; Nature; Peptide/MHC Complex; Peptides; Relative (related person); Role; Shapes; Side; Signal Transduction; Solutions; Spin Labels; Structure; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Techniques; base; design; dimer; insight

During the last funding interval as a merit award, this grant focused on the T cell receptor (TCR) complex composed of an αβ heterodimer and non-covalently associated CD3 signaling components. We continued crystallographic analysis with TCRs, providing the first structure of an αβ heterodimer in complex with a peptide/MHC (pMHC) class II ligand. In conjunction with a crystallographic structure of CD4 D1D2 binding to the same MHC class II molecule, a hydrophobic concavity formed by residues from membrane proximal α2 and β2 MHC class II domains was revealed. We excluded a direct TCRab-CD4 interaction, instead revealing how TCRαβ and CD4 signaling is coordinated in a "V-shape" around the antigenic peptide/pMHC class II complex. Solution structures of CD3εγ and CD3εδ ectodomain complexes were determined by NMR, uncovering for each dimer a unique side-to-side hydrophobic interface between their two Ig-like domains with parallel pairing of respective C-terminal β-strands, and suggesting how rigidified CD3 elements participate in TCR-based signal transduction. To now characterize the structural basis for early signal transduction events via this TCRαβ complex, four aims are proposed involving the CD3εγ heterodimer and associated TCR β chain. First, NMR-based methods will be used to determine the precise binding site on CD3εγ of activating and non-activating anti-CD3ε mAbs, given equivalent binding affinities. Second, CD3εγ-associated TCRβ chain recognition function during thymic development and T cell activation accounting for Vβ repertoire bias will be assessed. Third, the structure and function of the transmembrane (TM) segments of CD3ε and γ will be determined and their interaction with that of TCR β ascertained. Fourth, electron paramagnetic resonance (EPR) methods will be exploited to define the orientation and disposition of TM elements and how pMHC or anti-CD3ε mAbs affect TM depth, orientation and structural conformation. Distance measurements between the β chain and CD3ε and CD3γ ectodomains will also be examined before and after pMHC or anti- CD3 mAb ligation. That orthological rather than vertical force via CD3 components activates T cells suggests a dynamic mechanosensor TCR model; pMHC pulls on the TCR from the opposing antigen-presenting cell surface such that the αβ heterodimer then presses on the CD3 ectodomains to initiate signaling.

在作为优异奖的最后一个资助期间，这笔资助重点关注由 αβ 异二聚体和非共价相关的 CD3 信号成分组成的 T 细胞受体 (TCR) 复合物，我们继续对 TCR 进行晶体学分析，提供了 αβ 的第一个结构。与肽/MHC (pMHC) II 类配体复合的异二聚体与结合到相同 MHC II 类分子的 CD4 D1D2 的晶体结构相结合，形成疏水凹面。我们排除了直接的TCRab-CD4相互作用，而是揭示了TCRαβ和CD4信号如何在抗原肽/pMHC II类复合物周围以“V形”协调。通过 NMR 测定了 CD3εγ 和 CD3εδ 胞外域复合物的溶液结构，揭示了每个二聚体的两个 Ig 样结构域之间具有平行的独特的侧对侧疏水界面。各个 C 端 β 链的配对，并表明刚性 CD3 元件如何参与基于 TCR 的信号转导现在为了表征通过该 TCRαβ 复合物进行早期信号转导事件的结构基础，提出了涉及 CD3εγ 异二聚体和相关联的四个目标。首先，基于 NMR 的方法将用于确定激活和非激活抗 CD3ε mAb 在 CD3εγ 上的精确结合位点，前提是具有相同的结合亲和力。其次，将评估胸腺发育过程中 CD3εγ 相关的 TCRβ 链识别功能和导致 Vβ 库偏差的 T 细胞激活。第三，将确定 CD3ε 和 γ 跨膜 (TM) 片段的结构和功能以及它们之间的相互作用。第四，将利用电子顺磁共振（EPR）方法来确定TM元件的方向和配置以及pMHC或抗CD3ε单克隆抗体如何影响。 TM 深度、方向和结构构象还将在 pMHC 或抗 CD3 mAb 连接之前和之后检查 β 链与 CD3ε 和 CD3γ 胞外域之间的距离测量，通过 CD3 成分激活 T 细胞的直向力而不是垂直力。 TCR 模型；pMHC 从相反的抗原呈递细胞表面拉动 TCR，从而使 αβ 异二聚体压迫 CD3 胞外域发起信令。

项目成果

Plxnd1 expression in thymocytes regulates their intrathymic migration while that in thymic endothelium impacts medullary topology.

胸腺细胞中的 Plxnd1 表达调节其胸腺内迁移，而胸腺内皮中的 Plxnd1 表达则影响髓质拓扑。

• 发表时间: 2013
• 作者: Choi, Young I;Duke;Tan, Jing;Gui, Jingang;Singh, Manvendra K;Epstein, Jonathan A;Reinherz, Ellis L
• 通讯作者: Reinherz, Ellis L

Secondary immune amplification following live poliovirus immunization in humans.

人类活脊髓灰质炎病毒免疫后的二次免疫放大。

• 发表时间: 1987-09
• 作者: Hafler, D A;Fox, D A;Benjamin, D;Blue, M L;Weiner, H L
• 通讯作者: Weiner, H L

Surface molecules involved in self-recognition and T cell activation in the autologous mixed lymphocyte reaction.

自体混合淋巴细胞反应中参与自我识别和 T 细胞激活的表面分子。

• 发表时间: 1984-09
• 作者: Romain, P L;Schlossman, S F;Reinherz, E L
• 通讯作者: Reinherz, E L

Ta1, a novel 105 KD human T cell activation antigen defined by a monoclonal antibody.

Ta1，一种由单克隆抗体定义的新型 105 KD 人类 T 细胞激活抗原。

• 发表时间: 1984-09
• 作者: Fox, D A;Hussey, R E;Fitzgerald, K A;Acuto, O;Poole, C;Palley, L;Daley, J F;Schlossman, S F;Reinherz, E L
• 通讯作者: Reinherz, E L

Functional analysis of immunoreceptor tyrosine-based activation motif (ITAM)-mediated signal transduction: the two YxxL segments within a single CD3zeta-ITAM are functionally distinct.

基于免疫受体酪氨酸的激活基序 (ITAM) 介导的信号转导的功能分析：单个 CD3zeta-ITAM 内的两个 YxxL 片段在功能上不同。

• 发表时间: 1997-08
• 影响因子: 5.4
• 作者: Sunder;Lialios, F;Madsen, M;Koyasu, S;Reinherz, E L
• 通讯作者: Reinherz, E L