FUNCTIONAL CHARACTERIZATION OF FCER- AND FCER+ B CELLS

拟议研究的长期目标是更好地 了解B细胞亚群对人类免疫功能的影响 总体体液反应，并确定是否有明显的B 细胞亚群与自身免疫性疾病有关。目前， 我们对互联网的基本特征和功能的理解 各种B细胞亚群是有限的。此外，研究 这表明破坏性自身抗体主要来源于 一个特定的B细胞亚群仍然存在争议。部分内容 解决这些问题的困难在于缺乏 描述不同B细胞群体的明确标记。我们的 实验室发现，低亲和力的FceR可能相当有用 在这方面，由于它区分了传统的B细胞和Ly 1谱系和边缘区B细胞。相应地，具体的 该提案的目的是：1)确定特定的B是否 由FceR定义的细胞群体与 产生自身抗体。这将以谨慎的方式完成。 15株重组人B细胞亚群组成分析 来自NZB和SM/J亲本的近交系小鼠及其后代 当与NZW小鼠杂交时，这些RI线。基因 严重自身免疫性疾病的易感性已被隔离 在RI系之间，并在小鼠交配时变得明显 去新西兰。2)确定FceR+和FceR-B细胞 对多克隆刺激有不同的反应。3)确定是否 FceR+和FceR-B细胞对辅助性T细胞的不同应答 刺激物。4)确定TI-1和TI-2是否具有抗原特异性 应答主要来源于FceR-B细胞。5)更好 了解FceR+和FceR-B细胞的不同作用 试图获得有关FCER功能的新信息。 这将通过测试FceR，当交叉链接到 表面IgM或IGD可调节FceR+B细胞的活性。 将采用的方法包括广泛使用流式细胞术 用于分析和细胞分选，多克隆激活B细胞 在抗Ig、内毒素和淋巴因子的作用下，T细胞依赖的刺激 具有激活的TH1和TH2膜加上淋巴因子的B细胞，以及 半抗原TI-1和TI-1抗原特异性刺激B细胞的研究 2种抗原。这项建议与健康有关，这是由 从所有人都依赖于健康和 协调体液反应以产生保护性抗体。 B细胞室不能正常工作的不仅仅是 损害了我们对病原体的反应能力，但也可能导致 到破坏性的自身免疫性疾病，如类风湿性关节炎， 干燥综合征和系统性红斑狼疮。因此，它是 对于理解B细胞群体的复杂性至关重要， 以及导致自身抗体产生的条件。

The long term objective of the proposed research is to better understand the individual contributions of B cell subsets to the overall humoral response, and to determine whether a distinct B cell subset is associated with autoimmune disease. At present, our understanding of the basic characteristics and functions of the various B cell subsets is limited. Moreover, studies suggesting that damaging autoantibodies are primarily derived from a particular B cell subset remain controversial. Part of the difficulty in approaching these problems has been the lack of definitive markers to delineate different B cell populations. Our laboratory has found that the low affinity FceR may be quite useful in this regard, as it distinguishes conventional B cells from Ly 1 lineage and marginal zone B cells. Accordingly, the specific aims of the proposal are to: 1) Determine whether a particular B cell population, as defined by the FceR, is associated with the production of autoantibodies. This will be done by carefully analyzing the B cell subset composition of 15 lines of recombinant inbred mice derived from NZB and SM/J parents, and the progeny of these RI lines when crossed with NZW mice. The genetic predisposition for severe autoimmune disease has been segregated among the RI lines, and becomes manifest when the mice are mated to the NZW. 2) Determine whether FceR+ and FceR- B cells differentially respond to polyclonal stimuli. 3) Determine whether the FceR+ and FceR- B cells differentially respond to T helper cell stimuli. 4) Ascertain whether TI-1 and TI-2 antigen-specific responses are primarily derived from FceR-B cells. 5) Better understand the differing roles of FceR+ and FceR- B cells by attempting to gain new information as to the function of the FceR. This will be done by testing whether the FceR, when crosslinked to surface IgM or IgD, can modulated the activity of FceR+ B cells. The methods to be employed include extensive use of flow cytometry for analysis and cell sorting, polyclonal activation of B cells with anti-Ig, LPS and lymphokines, T cell dependent stimulation of B cells with activated TH1 and TH2 membranes plus lymphokines, and antigen-specific stimulation B cells with haptenated TI-1 and TI- 2 antigens. The health relatedness of this proposal is derived from the fact that all individuals depend upon a healthy and coordinated humoral response to produce protective antibodies. Failure of the B cell compartment to function properly not only compromises our ability to respond to pathogens, but can also lead to destructive autoimmune diseases such as rheumatoid arthritis, Sjorgren's syndrome and systemic lupus. Accordingly, it is essential to understand the complexity of the B cell population, and the conditions which lead to the production of autoantibodies.