Molecular mechanisms and function of P2X7 ion channel gating on T cells

P2X7是一种核苷酸门控离子通道，在炎症反应中发挥重要作用。该项目的主要目标是阐明T细胞上P2X7离子通道门控的分子机制和功能。为此，我们已经产生了纳米抗体(来自免疫大羊驼的单域抗体)，可以有效地阻断或加强细胞外核苷酸对P2X7的门控。这些纳米抗体提供了独特的工具来评估P2X7的表达，并在免疫反应过程中调节P2X7在小鼠和人类细胞上的功能。我们将使用P2X7突变体和嵌合体来确定P2X7的氨基酸残基，这些氨基酸残基对纳米体结合和受体的门控至关重要。我们将在两个小鼠模型中评估纳米体在实验性炎症中的治疗潜力。我们希望我们的研究能更好地理解P2X7在生理和病理免疫反应中的作用。我们将确定人类T细胞中P2X7的表达和功能，并使用我们的P2X7特异性纳米抗体来调节T细胞效应器的功能。这一结果有望为P2X7特异性纳米体的治疗应用提供基础。该项目的长期目标是将这些纳米体应用于改进炎症性疾病的诊断和治疗。

P2X7 is a nucleotide-gated ion channel that plays an important role in inflammation. The primary goal of the proposed project is to elucidate the molecular mechanisms and functions of P2X7 ion channel gating on T cells. To this end, we have generated nanobodies (single domain antibodies from immunized llamas) that effectively block or potentiate gating of P2X7 by extracellular nucleotides. These nanobodies provide unique tools to assess the expression of P2X7 and to modulate the function of P2X7 on murine and human cells during immune responses. We will use P2X7 mutants and chimeras to determine the amino acid residues of P2X7 important for nanobody binding and for gating of the receptor. We will evaluate the therapeutic potential of the nanobodies in experimental inflammation in two mouse models. We expect our studies to yield a better understanding of the role of P2X7 in physiological and pathological immune responses. We will characterize P2X7 expression and function in human T cells and use our P2X7-specific nanobodies to modulate T cell effector functions. The results are expected to provide a basis for therapeutical applications of P2X7-specific nanobodies. Long term goals of the project are to apply these nanobodies for improved diagnostics and therapy of inflammatory diseases.