Androgen Receptor in B Cell Development and Functions

描述(申请人提供)：性激素与免疫和炎症反应的调节有关。去势雄性小鼠会导致骨髓和外周血中B淋巴细胞的扩张。这种效果是由于男性荷尔蒙--雄激素的减少，雄激素通过激活雄激素受体(AR)起作用。去除AR还会导致骨髓和外周血中B细胞的扩张，这表明AR在B淋巴细胞的生成中起着负向调节作用。这项建议的总体目标是阐明AR调节B淋巴细胞生成的机制。拟议的研究将检验AR调节B前体细胞增殖和凋亡的假设。因此，去除骨髓细胞上的AR将增加B细胞的增殖和抗凋亡能力，导致B细胞的增殖，而B细胞的增加可能导致自身反应性B细胞的产生增加，从而导致自身免疫。为了验证这一假设，将为拟议的研究建立基质细胞和B细胞特异性AR基因敲除小鼠(S-ARKO和B-ARKO)。流式细胞术将测定各种骨髓B细胞在发育过程中的分布及其增殖率和凋亡率。还将测定自身抗体和自身反应性B细胞的产生。从G-Arko和B-Arko小鼠身上获得的结果将与它们的野生型后代进行比较，以确定AR在B淋巴细胞生成中的作用部位。我们将从S-ARKO、B-ARKO和野生型小鼠的纯化B细胞中提取RNA，以确定AR消融对细胞凋亡/增殖相关基因的影响，从而阐明AR的作用机制。最后，将研究和比较S-ARKO、B-ARKO和野生型对诱导自身免疫性疾病的易感性。这些研究结果将为阐明AR在调节B淋巴细胞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.