CD1d-restricted T cells and anti-phospholipid antibodies

描述（由申请人提供）：抗磷脂抗体是一种与显著心血管病变、中风和复发性流产相关的自身抗体。目前尚不清楚这些抗体是如何产生的，或者T细胞是否参与了它们的产生。CD 1d限制性T细胞是一种新型的T淋巴细胞群体，其识别脂质和糖脂抗原，包括在某些情况下的磷脂。CD 1d分子在抗原呈递细胞（包括B细胞）上表达。该提案研究了B细胞的抗磷脂抗体产生可以由CD 1d限制性T细胞以抗原特异性方式调节的假设。该分析的重点是致病性抗磷脂抗体水平升高的存在是否与CD 1d限制性T细胞的数量、激活状态、功能或抗原特异性的变化相关。这将在以下具体目的中进行评估：i）使用负载脂质抗原的CD 1d四聚体进行流式细胞术分析，以检测和分析来自具有抗磷脂抗体的患者的外周血的CD 1d限制性T细胞的功能特性，与对照供体相比; ii）使用CD 1d-Fc融合蛋白和荧光或放射性标记的磷脂分析不同磷脂与CD 1d分子的结合; iii）CD 1d限制性T细胞克隆将来源于具有抗磷脂抗体的患者和对照供体，并用于研究对单个磷脂的反应性，并测试产生抗磷脂抗体的B细胞的识别。这项研究将提供新的见解的作用，自身反应性T细胞在自身抗体的产生，并可能提供新的诊断和治疗方法与致病性抗磷脂抗体相关的条件。

DESCRIPTION (provided by applicant): Anti-phospholipid antibodies are a type of autoantibody associated with significant cardiovascular pathology, stroke, and recurrent miscarriages. It is not known how these antibodies arise, or whether T cells are involved in their generation. CD1d-restricted T cells are a novel population of T lymphocytes that recognize lipid and glycolipid antigens, including in some cases phospholipids. CD1d molecules are expressed on antigen presenting cells, including B cells. This proposal investigates the hypothesis that anti-phospholipid antibody production by B cells could be regulated by CD1d-restricted T cells in an antigen specific manner. The analysis focuses on whether the presence of elevated levels of pathogenic anti-phospholipid antibodies correlates with changes in the numbers, activation states, functions, or antigen specificities of CD1d-restricted T cells. This will be assessed in the following specific aims: i) flow cytometric analysis will be performed using lipid antigen loaded CD1d tetramers to detect and analyze the functional properties of CD1d-restricted T cells from peripheral blood of patients with anti-phospholipid antibodies compared to control donors; ii) binding of different phospholipids to CD1d molecules will be analyzed using CD1d-Fc fusion proteins and fluorescent or radiolabeled phospholipids; iii) CD1d-restricted T cell clones will be derived from patients with anti-phospholipid antibodies and control donors, and used to investigate reactivity to individual phospholipids, and to test recognition of anti-phospholipid antibody producing B cells. This investigation will provide new insight into the role of autoreactive T cells in the generation of autoantibodies, and may provide new diagnostic and therapeutic approaches for conditions associated with pathogenic anti-phospholipid antibodies.