Creation of a conditional IL-6 knockout mouse

描述（由申请人提供）：脓毒症是危重患者死亡和发病的主要原因。我们已经使用小鼠模型（盲肠结扎和穿刺或CLP）研究了一段时间，并发现了几个器官系统中的一些重要异常，特别是肝脏，肺和心脏。我们发现的关键是一种重要的炎症介质细胞因子白细胞介素（IL）-6的活性丧失。我们已经将我们的研究扩展到先天性缺乏IL-6的小鼠（IL-6 -/-或“敲除”小鼠）。我们在野生型（IL-6 +/+）小鼠中观察到的所有异常在IL-6 -/-动物中更严重。然而，在IL-6 -/-小鼠中，在受孕时产生敲除。因此，代偿途径可以发展。此外，IL-6 -/-小鼠中IL-6的损失并不能真正模拟野生型败血症中发生的情况。急性消除IL- 6的能力将在研究脓毒症中非常有用。不幸的是，目前消除IL-6的技术具有显著的缺点。IL-6 -/-小鼠的使用受到限制，因为无法将具有先天性出生缺陷的动物中的发现外推到急性情况。使用针对IL-6的抗体可有效地从循环中消除IL-6，但不能从组织中消除IL-6，脓毒症中的大多数损伤发生在组织中。在这篇文章中，我们提出了另一种方法。我们将产生和繁殖一只小鼠，从中可以急性消除IL-6。这将通过创建IL-6构建体基因来实现，所述IL-6构建体基因两侧是可以被称为cre重组酶的酶切割的两个位点。将该构建体引入生殖组织，并繁殖含有该序列的小鼠。一旦我们建立了一个动物系，我们将能够通过将表达cre重组酶的腺病毒载体施用到气管或血流中来从肺和肝中消除IL-6。这将允许从肺和肝急性消除IL-6。这种工具的可用性将使我们能够更好地研究IL-6在脓毒症诱导的这些器官变化的发病机制中的作用。这将增加对脓毒症病理生理学的理解。脓毒症是导致死亡和发病的重要原因。据估计，每年用于治疗这种疾病的费用超过160亿美元。因此，脓毒症构成主要的公共卫生问题。.不幸的是，这种疾病的病理生理学知之甚少。更好地了解这种疾病与美国公民的护理极其相关。

DESCRIPTION (provided by applicant): Sepsis is a major cause of mortality and morbidity in the critically ill. We have been studying the disorder using a murine model (cecal ligation and puncture or CLP) for some time and have uncovered some important abnormalities in several organ systems, notably liver, lung and heart. Key among our findings is a loss in the activity of an important inflammatory mediator, the cytokine interleukin (IL) - 6. We have extended our investigations to mice with a congenital absence of IL-6 (IL-6 -/- or "knockout" mice). All the abnormalities we observed in wild type (IL-6 +/+) mice were worse in IL-6 -/- animals. However, in IL-6 -/- mice, the knockout is generated at conception. Therefore, compensatory pathways can develop. In addition, the loss of IL-6 in IL-6 -/- mice does not truly mimic what occurs in sepsis in wild types. The ability to acutely eliminate IL- 6 would be extremely useful in studying sepsis. Unfortunately, current techniques to eliminate IL-6 have significant drawbacks. Use of IL-6 -/- mice is limited by an inability to extrapolate findings in animals with a congenital, in-born error to the acute situation. Use of antibodies directed at IL-6 is effective in eliminating IL-6 from the circulation but not from tissues, where most of the damage in sepsis occurs. In this grant, we propose another approach. We will generate and breed a mouse from which IL-6 can be acutely eliminated. This will be accomplished by creating an IL-6 construct gene that is flanked by two sites that can be cleaved by an enzyme called cre-recombinase. This construct will be introduced into germ tissue and a mouse containing this sequence will be bred. Once we have established a line of animals, we will be able to eliminate IL-6 from lung and liver by administering an adenoviral vector expressing cre-recombinase into the trachea or the bloodstream. This will allow acute elimination of IL-6 from lung and liver. The availability of such a tool will allow us to better investigate that role of IL-6 in the pathogenesis of sepsis-induced changes in these organs. This will provide increased understanding op septic pathophysiology. Sepsis is an important cause of mortality and morbidity. It has been estimated that in excess of $16 billion are spent each year in the use to treat this disorder. As such, sepsis constitutes a major public health problem. . Unfortunately, the pathophysiology of the disease is poorly understood. A better understanding of this disease is extremely relevant to the care of the citizens of the United States.