Androgen Receptor in B Cell Development and Functions

描述（由申请人提供）：性激素与免疫和炎症反应的调节有关。雄性小鼠的阉割导致骨髓和外周血中B淋巴细胞的扩增。这种影响是由于男性性激素雄激素的减少，雄激素通过雄激素受体（AR）的激活起作用。小鼠AR的消融也导致骨髓和外周血中B细胞的扩增，提示AR在B淋巴生成中起负调节作用。本提案的总体目标是阐明AR调节B淋巴生成的机制。本研究将验证AR调控B前体细胞增殖和凋亡的假说。因此，骨髓细胞中AR的消融会增加B细胞的增殖和对凋亡的抵抗，导致B细胞扩增，而B细胞的增加可能导致自身反应性B细胞的产生增加，从而产生自身免疫。为了验证这一假设，将在拟议的研究中产生基质细胞和B细胞特异性AR敲除（S-ARKO和B- arko）小鼠。流式细胞术将测定骨髓中各种B细胞在发育过程中的分布及其增殖和凋亡率。自身抗体和自身反应性B细胞的产生也将被确定。G-ARKO和B- arko小鼠的结果将与野生型小鼠进行比较，以确定AR在B淋巴生成中的作用部位。我们将从S-ARKO、B- arko和野生型小鼠纯化的B细胞中分离RNA，鉴定AR消融影响凋亡/增殖的相关基因，从而阐明AR作用的机制。最后，我们将研究和比较S-ARKO、B-ARKO和野生型对自身免疫性疾病诱导的易感性。这些研究结果将为阐明AR在调节b淋巴生成和自身免疫中的作用及其作用机制，以及更好地理解和治疗自身免疫性疾病提供有用的信息。

DESCRIPTION (provided by applicant): Sex hormones are linked to regulation of immune and inflammatory responses. Castration of male mice results in expansion of B lymphocytes in the bone marrow and peripheral blood. This effect is due to reduction of male sex hormone, androgen, which acts through activation of androgen receptor (AR). Ablation of AR in mice also leads to expansion of B cells in the bone marrow and peripheral blood, suggesting that AR plays a negative regulatory role in B lymphopoiesis. The overall objective of this proposal is to elucidate the mechanism through which AR regulates B lymphopoiesis. The proposed studies will test the hypothesis that AR regulates proliferation and apoptosis of B precursor cells. Thus, ablation of AR from bone marrow cells will increase proliferation and resistance to apoptosis of B cells leading to B cell expansion, whereas increased B cells might result in increased production of self-reactive B cells hence autoimmunity. To test this hypothesis, stromal cell and B cell specific AR knockout (S-ARKO and B-ARKO) mice will be generated for the proposed studies. The distribution of various bone marrow B cells along the developmental pathway and their proliferating and apoptosis rates will be determined by flow cytometry. The production of autoantibody and self-reactive B cell also will be determined. The results obtained from G-ARKO and B-ARKO mice will be compared with their wild type littermates to determine the sites of AR action in B lymphopoiesis. RNA will be isolated from purified B cells of S-ARKO, B-ARKO, and wild type mice to identify the genes related to apoptosis/proliferation affected by AR ablation so that the mechanism of AR action can be elucidated. Finally, the susceptibility of S-ARKO, B-ARKO, and wild type to induction of autoimmune disease will be studies and compared. The results of the proposed studies will provide useful information toward elucidation of the role of AR and its mechanism of action in regulating B-lymphopoiesis and autoimmunity, and better understanding and treatment of autoimmune diseases.