Pro-resolving lipid mediators in mycobacterial infection

描述（由申请人提供）：本提案探讨了促溶解脂质介质，溶解蛋白，保护蛋白和蛋白在分枝杆菌发病机制中的作用。这些最近发现的脂质有效地对抗无菌炎症并促进损伤组织的修复，但它们在感染引起的炎症中的作用仍未得到充分研究。结核病的严重程度可因两种截然相反的状态而增加：炎症过少或过多。在人类和marinum感染的斑马鱼（结核模型）中，白三烯A4羟化酶（LTA4H）活性高和低的斑马鱼感染严重程度增加。这是由于促炎白三烯B4的过量产生，在具有高LTA4H基因型的患者中引起过量的TNF和炎症。在低lta4h基因型中，过量产生抗炎脂素A4导致TNF不足，抑制保护性炎症。在Aim 1中，本提案将首次使用质谱法识别斑马鱼中的溶解蛋白、保护蛋白和蛋白，并确定这些脂质是否在对海洋支原体感染的反应中受到调节。为了证明这些脂质在斑马鱼和哺乳动物中的相似功能，将分析它们抑制中性粒细胞募集和TNF产生的能力。在Aim 2中，将使用morpholino技术和注射LTA4H转录物来产生低LTA4H和高LTA4H的斑马鱼。感染后，将使用促溶解脂质介质来测量这些脂质对每种基因型宿主的影响。如果促溶解脂质介质可以减轻高LTA4H斑马鱼的海洋分枝杆菌感染，那么这些脂质具有很大的潜力，可以作为结核病的新疗法，因为一种稳定的溶解蛋白类似物目前正在II期临床试验中。这些目标的完成不仅有助于我们理解免疫病理与分枝杆菌发病机制的相互作用，而且还可能为具有免疫病理成分的疾病提供新的治疗方法。

DESCRIPTION (provided by applicant): This proposal examines the role of the pro-resolving lipid mediators, resolvins, protectins, and maresins, in mycobacterial pathogenesis. These recently discovered lipids effectively combat sterile inflammation and promote repair of injured tissue, but their role in inflammation caused by infection remains largely unexplored. Severity of tuberculosis can be increased by two diametrically opposed states: too little inflammation or too much. In both humans and M. marinum-infected zebrafish, a model for tuberculosis, those with high and low activity of leukotriene A4 hydroxylase (LTA4H) had increased infection severity. This was due to excessive production of pro-inflammatory leukotriene B4,causing excess TNF and inflammation in those with the high- LTA4H genotype. In the low-LTA4H genotype, excessive production of anti-inflammatory lipoxin A4 caused inadequate TNF, inhibiting protective inflammation. In Aim 1, this proposal will identify resolvins, protectins, and maresins for the first time in zebrafish using mass spectrometry and determine if these lipids are regulated in response to M. marinum infection. To demonstrate similar functions of these lipids in zebrafish and mammals, their ability to inhibit neutrophil recruitment and TNF production will be assayed. In Aim 2, low- LTA4H and high-LTA4H zebrafish will be generated using morpholino technology and injection of LTA4H transcript. After infection, pro-resolving lipid mediators will be administered to measure the effect of these lipids on hosts of each genotype. If pro-resolving lipid mediators attenuate M. marinum infection in high- LTA4H zebrafish, these lipids have great potential as new treatments for tuberculosis, as a stabilized resolvin analogue is currently in Phase II clinical trials. Completion of these Aims will not only contribute to our understanding of how immunopathology is interwoven with mycobacterial pathogenesis, but also potentially suggest new treatments for diseases with an immunopathology component.