CD1d-restricted T cells and anti-phospholipid antibodies

说明(申请人提供)：抗磷脂抗体是一种与重大心血管病变、中风和反复流产有关的自身抗体。目前尚不清楚这些抗体是如何产生的，也不知道T细胞是否参与了它们的生成。CD1d限制性T细胞是一类识别脂类和糖脂抗原的新型T淋巴细胞，在某些情况下包括磷脂。CD1d分子表达于抗原提呈细胞，包括B细胞。这项建议探讨了B细胞产生抗磷脂抗体的假设，即CD1d限制的T细胞可以抗原特异性的方式调节B细胞产生抗磷脂抗体。分析的重点是致病性抗磷脂抗体水平的升高是否与CD1d限制性T细胞的数量、激活状态、功能或抗原特异性的变化有关。这项工作将在下列特定目标中进行评估：i)使用脂类抗原负载的CD1d四聚体进行流式细胞术分析，以检测和分析来自抗磷脂抗体阳性患者和对照捐赠者外周血中CD1d限制性T细胞的功能特性；ii)使用CD1d-Fc融合蛋白和荧光或放射性标记磷脂分析不同磷脂与CD1d分子的结合；iii)CD1d限制性T细胞克隆将来自抗磷脂抗体阳性患者和对照捐赠者，并用于研究其对单个磷脂抗体的反应性，以及测试抗磷脂抗体产生B细胞的识别能力。这项研究将为自身反应性T细胞在自身抗体产生中的作用提供新的见解，并可能为与致病性抗磷脂抗体相关的疾病提供新的诊断和治疗方法。

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.