The Mechanism of Co-Receptor Regulation of B-cell Activation
The Mechanism of Co-Receptor Regulation of B-cell Activation
批准号:
8745432
负责人:
Susan Pierce
金额:
$62.92万
依托单位国家:
美国
项目类别:
财政年份:
--
资助国家:
美国
项目状态:
未结题
起止时间:
至
关键词:
AffinityAntibodiesAntibody FormationAntigen-Antibody ComplexAntigensAttentionAutoimmune DiseasesAutoimmunityB-Cell ActivationB-Cell NeoplasmB-LymphocytesBindingCD19 geneCoinCollaborationsComplementComplement 3d ReceptorsComplexCoupledCytoplasmic TailDimerizationEnvironmentEventFc ReceptorFluorescence Resonance Energy TransferHumanImageImaging technologyImmune responseImmunoglobulin GImmunoglobulin MImmunologic MemoryLifeLipidsMediatingMembraneMembrane MicrodomainsMemoryMemory B-LymphocyteMicroscopicMicroscopyMolecularPathway interactionsPhosphotransferasesPlayProcessRecruitment ActivityRegulationResolutionRestRoleSignal PathwaySignal TransductionSpatial Distributionantigen bindingcellular imagingkillingsmolecular sizereceptorresponsesingle moleculespatial relationship
中文摘要
B细胞对抗原的反应是由多种共受体调节的,这些共受体向B细胞传递有关抗原质量和正在进行的免疫反应状态的信息。我们使用高分辨率活细胞成像来描绘B细胞活化中非常早期抗原驱动事件的进展,为评估协受体的影响提供了新的背景。利用高分辨率荧光共振能量转移(FRET)结合全内反射显微镜(TIRFM)和单分子跟踪,我们提供了一个有序的过程发生在几秒到几分钟的BCR结合抗原的证据。抗原结合的bcr形成固定的簇,然后通过分子诱捕增大大小。这些团簇扰乱了局部脂质环境,导致脂筏在BCR团簇周围聚集。由于膜的扰动,途径中的第一个激酶,Lyn,被筏脂拴在膜上,被带到靠近BCR簇的分子附近。同时,Lyn磷酸化了BCR的Ig α - β细胞质结构域,并且Ig α - β链经历了从封闭到开放形式的构象变化。Syk被招募到磷酸化的BCR中,信号级联被触发。
英文摘要
The B cell response to antigen is regulated by a variety of co-receptors that convey information to the B cell about the quality of the antigen and the status of the ongoing immune response. Our progress using high resolution live cell imaging to delineate the very early antigen driven events in B cell activation has provided a new context in which the impact of coreceptors can be evaluated. Using high resolution fluorescence resonance energy transfer (FRET) coupled with total internal reflection microscopy (TIRFM) and single molecule tracking we provided evidence for an ordered process that occurs within seconds to minutes of the BCR binding antigen. Antigen bound BCRs form immobile clusters that then grow in size by molecular trapping. The clusters perturb the local lipid environment causing lipid rafts to coalesce around the BCR clusters. As a consequence of the membrane perturbation the first kinase in the pathway, Lyn, that is tethered to the membrane by raft lipids is brought into close molecular proximity to the BCR clusters. Simultaneously, Lyn phosphorylates the Ig alpha beta cytoplasmic domain of the BCR and the Ig alpha beta chains undergo a conformational change from a closed to an open form. Syk is recruited to the phosphorylated BCR and the signaling cascades are triggered.
Over the last year we characterized the effect of the inhibitory receptor FcgammaRIIB engagement on the activation of human naive and MBCs. Using high-resolution imaging we showed that human MBCs are more robust than naive B cells at each step in the initiation of BCR signaling, including interrogation of antigen containing membranes, formation of sub-microscopic BCR oligomers and recruitment and activation of signaling-associated kinases. Despite their robust response to antigen, MBCs remain highly sensitive to FcgammaRIIB-mediated inhibition. These observations are important for understanding the regulation of memory antibody responses.
We also demonstrated in human B cells that the rapid accumulation of BCRs at the interface of the B cell on antigen-containing bilayer triggers the independent accumulation of phosphorylated CD19 accompanied by the accumulation and colocalization of phosphorylated PI3K, a key component in the BCR-CD19 signaling pathway. Remarkably, coligation of the BCR and FcgammaRIIB blocked the accumulation of the BCR, and phosphorylated CD19 and PI3K. These results suggest an additional facet of the inhibition of BCR signaling by the FcgammaRIIB. We will continue these studies in collaboration with Dr. Ezio Bonvini and his colleagues at Macrogenics. Dr. Bonvinis group has developed bi-functional antibodies coined DARTS that allow the dimerization of the BCR and CD19 or the BCR and FcgammaRIIB. The BCR-FcgammaRIIB DARTS are showing efficacy in blocking B cell activation in autoimmune disease and killing BCR-dependent B cell tumors. We are exploring the effect of these DARTS on BCR signaling using high resolution live cell imaging.
Over the last year we have made progress in establishing super-high resolution imaging to describe, at the 10-50 nm level, the spatial relationship between the BCR, CD19 and FcgammaRIIB in resting human B cells and in B cells in which the BCR is ligated alone by Ag or Fcgamma is ligated by immune complexes. We are also characterizing the spatial distribution of these receptors on naive human IgM expressing B cells and in IgG-expressing memory B cells.
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会议论文
B Cell Biology
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批准号:10272086
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项目类别:
-
资助金额:$304.22万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Membrane Microdomains And B Cell Signaling
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批准号:6521525
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Intracellular Trafficking Of The B cell Antigen Receptor
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批准号:6521528
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
The Mechanism of Co-Receptor Regulation of B-cell Activation
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批准号:8555905
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项目类别:
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资助金额:$48.65万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Membrane Microdomains And B- Cell Signaling
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批准号:7196688
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
B cell Receptor Dysregulation in Cancer and Autoimmune Disease
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批准号:8745551
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项目类别:
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资助金额:$25.17万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Intracellular Trafficking and Signaling Of The B-cell Antigen Receptor
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批准号:8745390
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项目类别:
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资助金额:$62.92万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
The Mechanism of Co-Receptor Regulation of B-cell Activation
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批准号:9566642
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项目类别:
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资助金额:$46.83万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
B cell Receptor Dysregulation in Cancer and Autoimmune Disease
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批准号:8157106
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项目类别:
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资助金额:$27.22万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
The Mechanism of Co-Receptor Regulation of B-cell Activation
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批准号:8156981
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项目类别:
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资助金额:$40.84万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Membrane Microdomains And B- Cell Signaling
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批准号:6987011
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Intracellular Trafficking and Signaling Of The B-cell Antigen Receptor
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批准号:9563890
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项目类别:
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资助金额:$134.24万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
B cell Receptor Dysregulation in Cancer and Autoimmune Disease
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批准号:8556031
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项目类别:
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资助金额:$19.43万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Membrane Microdomains And B- Cell Signaling
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批准号:6669902
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
B Cell Biology
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批准号:10692071
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项目类别:
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资助金额:$249.73万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Malaria immunology
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批准号:10692120
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项目类别:
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资助金额:$246.44万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Intracellular Trafficking and Signaling Of The B-cell Antigen Receptor
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批准号:7732566
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项目类别:
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资助金额:$32.17万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
The Initiation of B-Cell Signaling
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批准号:8946353
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项目类别:
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资助金额:$88.91万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
The Initiation of B-Cell Signaling
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批准号:8156932
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项目类别:
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资助金额:$108.89万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
Malaria immunology
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批准号:10272145
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项目类别:
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资助金额:$201.66万
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财政年份:--
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负责人:Susan Pierce
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依托单位:
海外基金