Effects and mechanisms of cold-induced stress on the development of Chlamydia muridarum genital infection in a mouse model

项目主任/首席调查员(最后、第一、中间)：贝莱，特斯法耶 项目摘要 沙眼衣原体生殖道感染是世界上最常见的性传播细菌疾病。如果离开 如果不治疗，衣原体生殖器感染会导致盆腔炎、宫外孕和不孕。它是广泛存在的 知道对这种疾病的免疫病理反应，而不是感染本身，仍然是一个严重的公共卫生问题 有问题。了解压力如何影响对性行为的抵抗力或易感性是有医学意义的。 传播疾病。许多研究人员研究了沙眼衣原体的免疫学反应，但沙眼衣原体的影响 对衣原体生殖器感染的压力尚未被探索，也不被理解。我们的研究试图理解 他们之间的关系。为了做到这一点，我们开发了一种通过每天浸泡小鼠的慢性应激模型。 每天在冷水中浸泡5分钟，连续21天。这种冷诱导应激(CIS)小鼠模型显示强度增加 沙眼衣原体生殖器感染与应激激素去甲肾上腺素升高有关 (Ne)在初次感染和继发感染期间。我们的研究表明，独联体促进增加β-2- 肾上腺素能受体(β2-AR)在CD4T细胞中的表达以及辅助性T细胞(Th2)的分化，特别是通过 增加GATA-3的表达和IL-4的分泌。此外，我们还发现β1/β2-AR KO对 鼠疫杆菌生殖器感染比WT C57BL/6J株强。这些观察结果促使我们使用β2-AR KO模型 探讨衣原体生殖道感染的免疫发病机制。我们最近发现，β2-AR KO对 比WT C57BL/6J.然而，慢性顺应性鼻炎感染的潜在机制和 我们的小鼠模型的免疫发病机制尚不清楚。这项提案的中心目标是定义 应激抑制免疫系统增强生殖器线虫感染强度的机制(S) 小鼠感染及其免疫病理机制。我们假设独联体产生的去甲肾上腺素导致刺激 小鼠𝛽𝛽2-AR信号通路进而抑制对小鼠生殖器官的保护性免疫反应 感染。为了验证这一假设，我们将进行以下三个实验作为该项目的一部分：在目标1中，我们 将比较应激β2-AR-/-小鼠和C57BL/6J(β2-AR/)小鼠的CD4T细胞亚群免疫反应。我们 推测Th1型受体缺陷可促进应激小鼠β细胞因子产生的恢复，这可能是 可使鼠疫霉菌清除。在Aim#2中，我们将探索β2-AR的下游信号通路 应激KO小鼠与应激C57BL/6J小鼠的比较。获得的数据可能为深入了解β2-AR的参与提供依据 在cAMP-蛋白激酶A(PKA)和丝裂原活化蛋白激酶(MAPK)途径中抑制免疫 系统。在目标3中，我们将评估应激β2-AR KO与WT C57BL/6J小鼠相比的组织病理学变化， 然后我们将评估细胞因子在病理学中的作用。来自该方法的数据将表明， β2-AR KO小鼠的输卵管病理学。拟议中的实验将提供对免疫抑制的洞察。 衣原体免疫发病中内分泌和免疫系统相互作用的机制。 总体而言，从这些β2-AR缺陷小鼠获得的结果，结合我们之前的发现和其他人的发现，可以 揭示了慢性压力对人类衣原体疾病严重程度的影响的更多证据。 OMB编号0925-0001/0002(批准的第03/2020版至2023年2月28日)续页格式页

Program Director/Principal Investigator (Last, First, Middle): Belay, Tesfaye Project Summary Chlamydia trachomatis genital infection is the most common sexually-transmitted bacterial disease in the world. If left untreated, chlamydia genital infection leads to pelvic inflammatory disease, ectopic pregnancy, and infertility. It is widely known that the immunopathological reactions to this disease, rather than the infection itself, remain a serious public health problem. There is medical interest in understanding how stress may impact resistance or susceptibility to sexually transmitted diseases. Many investigators have examined the immunological responses to C. trachomatis, but the effect of stress on chlamydia genital infection has not yet been explored and is not understood. Our research seeks to understand the relationship between them. To accomplish this, we have developed a chronic-stress model in mice by daily immersing mice in cold water for five minutes daily for 21 days. This cold-induced stress (CIS) mouse model shows the increased intensity of Chlamydia muridarum genital infection, which is associated with the elevation of the stress hormone norepinephrine (NE) during both primary and secondary infections. Our research has demonstrated that CIS promotes increased beta2- adrenergic receptor (β2-AR) expression in CD4 + T cells, as well as T helper 2 (Th2) differentiation, specifically by increasing the expression of GATA-3 and IL-4 secretion. Moreover, we showed that β1/β2-AR KO are less susceptible to C. muridarum genital infection than the WT C57BL/6J. Those observations prompted us to use a β2-AR KO model and explore the immunopathogenesis of chlamydia genital infection. We recently found that the β2-AR KO is more resistant to C. muridarum than the WT C57BL/6J. However, the underlying mechanisms of chronic CIS on the infection and the immunopathogenesis in our mouse model remain unknown. The central objective of this proposal is to define the mechanism(s) by which stress may suppress the immune system and increase the intensity of genital C. muridarum infection and its immunopathologies in mice. We hypothesize that the NE produced by CIS leads to stimulation of the 𝛽𝛽2-AR signaling pathway in mice which, in turn, suppresses the protective immune response against C. muridarum genital infection. To test this hypothesis, we will conduct the following three experiments as part of this project: In Aim #1, we will compare the CD4+ T cell subset immune responses of stressed β2-AR-/- mice and C57BL/6J (β2-AR+/+) mice. We anticipate that the deficiency of β2-AR enhances the restoration of Th1 cytokine production in stressed mice, which may result in the clearance of C. muridarum. In Aim #2, we will explore the downstream signaling pathways from the β2-AR of stressed KO mice compared to stressed C57BL/6J mice. Data obtained may provide insight into the involvement of β2-AR in cAMP-protein kinase A (PKA) and mitogen-activated protein kinase (MAPK) pathways in suppressing the immune system. In Aim#3, we will assess the histopathological changes of stressed β2-AR KO as compared to WT C57BL/6J mice, and we will then evaluate the contributions of cytokines to pathology. Data from this method will demonstrate a reduced oviductal pathology in β2-AR KO mice. The proposed experiments will provide insight into the immunosuppression mechanisms resulting from the interactions of the endocrine and immune systems in the immunopathogenesis of chlamydia. Overall, results obtained from these β2-AR deficient mice, combined with our previous findings and those of others, could reveal more evidence of the effects of chronic stress on the severity of chlamydial disease in humans. OMB No. 0925-0001/0002 (Rev. 03/2020 Approved Through 02/28/2023) Page Continuation Format Page