Defining the role of sphingolipids on Porphyromonas gingivalis outer membrane vesicle uptake and elicited inflammation

摘要 在培训计划中概述了这项NRSA个人博士前奖学金，以促进拟议的工作 健康相关研究（F31）奖的多样性将支持第四年博士前候选人Zavier Eure 在佛罗里达大学的研究生课程中，厄尔先生在他的工作中表现出了非凡的卓越。 学者和他的新的实验室技术的学习，而在UF BMS计划，目前支持 口腔生物学系R90/T90培训补助金。厄尔先生的研究和培训计划将提供 他的技能需要一个剧目录取到他的博士前培训的下一个层次，也有机会 以进一步推进其职业生涯和专业发展。获得F31将使厄尔先生有能力 为了实现他成为科学界领导者的长期目标，并为该领域做出重大贡献， 生物医学研究。 牙周病是一种高度流行的慢性炎症性疾病，在美国， 超过30岁牙周病是精心策划的各种微生物组成的龈下 微生物群在易感个体中，龈下微生物群的生态失调促进了牙龈上皮细胞的失调。 导致支撑牙齿的软硬组织不可逆转的破坏的炎症。 牙龈卟啉单胞菌（Porphyromonas gingivalis，Pg）是一种口腔细菌，通常与微生物生态失调有关， 牙周病PG被认为是致病菌，因为它可以操纵宿主的炎症反应。 然而，在健康状态期间，Pg也可以保留在口腔中。的 Pg如何促进炎症环境变化的各种机制尚不清楚。 鉴定 我们的实验室已经发现Pg鞘脂（SL）通过外部途径限制宿主炎症和宿主细胞SL转移， 膜囊泡是Pg的潜在转运机制。宿主细胞摄取OMV的机制是 已知影响引发的炎症和OMV成分（SL+或SL-）在炎症途径中发挥重要作用 OMV摄取。髓样分化因子88（MyD 88）也已知参与先天性免疫感应， 的Pg OMV，并已被发现是一个目标的免疫调节的Pg。我假设Pg SL促进 限制炎症的OMV摄取机制与SL促进免疫调节机制相关 针对MyD 88。我计划使用THP-1人类细胞系和原代人类巨噬细胞来表征 目的1：研究含SL的Pg OMV的摄取机制，并确定MyD 88在Pg SL介导的OMV摄取中的作用。 免疫调节（目的2）。

ABSTRACT The proposed work outlined in the training plan for this NRSA Individual Predoctoral Fellowships to Promote Diversity in Health-Related Research (F31) Award will support fourth year pre-doctoral candidate Zavier Eure throughout his graduate program at University of Florida. Mr. Eure has displayed phenomenal excellence in his academics and his learning of new lab techniques while being in the UF BMS program and currently supported by the Oral Biology Department R90/T90 Training Grant. The research and training plan for Mr. Eure will provide him a repertoire of skills needed to matriculate to the next level of his pre-doctoral training and also opportunities to further advance his career and professional development. Being awarded this F31 will offer Mr. Eure the ability to achieve his long-term goal of becoming a leader in science and provide a significant contribution to the field of biomedical research. Periodontal disease is a highly prevalent chronic inflammatory disease affecting up to 42% of adults in the US over the age of 30. Periodontal disease is orchestrated by various microorganisms that make up the subgingival microbiota. In susceptible individuals, dysbiosis of the subgingival microbiota promotes a dysregulated inflammation that causes an irreversible destruction of the soft and hard tissues supporting the teeth. Porphyromonas gingivalis (Pg) is an oral bacterium commonly associated with the microbial dysbiosis leading to periodontal disease. Pg is referred to as a pathobiont because its known to manipulate the host inflammatory milieu to facilitate disease states, however Pg can also remain in the oral cavity during healthy states. The various mechanisms behind how Pg contributes to the changing inflammatory milieu is yet to be identified. Our lab has discovered Pg sphingolipids (SLs) limit host inflammation and host cell SL transfer via outer membrane vesicles is a potential transport mechanism for Pg. The mechanism of OMV uptake by host cells is known to impact elicited inflammation and OMV composition (SL+ or SL-) plays an important role in the route for OMV uptake. Myeloid differentiation factor 88 (MyD88) is also known to be involved in the innate immune sensing of Pg OMVs and has been found to be a target for immunomodulation by Pg. I hypothesize Pg SLs facilitates an OMV uptake mechanism that limits inflammation and that SLs promote an immunomodulatory mechanism linked to targeting of MyD88. I plan to use the THP-1 human cell line and primary human macrophages to characterize the uptake mechanism of SL-containing Pg OMVs (Aim 1) and to determine the role of MyD88 in Pg SL-mediated immunomodulation (Aim 2).