Decoding vomocytosis for cell-medaited, intra-lymph nodal delivery of microparticle vaccines

细胞介导的淋巴结内递送微粒疫苗的解码细胞吞噬作用 摘要/总结 加州大学戴维斯分校的免疫调节生物材料实验室专注于开发新型 可以操纵免疫系统的生物材料系统。我们的目标是设计下一代 用于免疫相关疾病的免疫治疗剂。这一多学科工作包括 生物材料工程、药物输送、免疫学、生物化学和细胞生理学方面。之一 我们实验室的主要研究项目是了解颗粒的控制性呕吐（非溶解性胞吐） 吞噬细胞的物质。我们阐明这一过程的主要动机是开发一种“智能”的 可以实现吞噬细胞介导的疫苗直接递送至淋巴的微粒系统 节点驻留的细胞。吞噬细胞（特别是巨噬细胞[MΦ]和树突状细胞[DC]）已经进化为 通过外周组织循环，吞噬外来颗粒物质并运输到淋巴结， 摄取以将关于外周环境的信息传递给适应性免疫系统的细胞因子。 显然，吞噬细胞可以将颗粒物质从外周组织转运到淋巴器官。 典型地，在吞噬作用之后，并且当吞噬细胞穿过淋巴管时，具有以下性质的物质： 被吞噬的细胞被吞噬体（含有被吞噬的细胞的细胞质中的空泡）吸收。 颗粒），其中由于活性氧物质（ROS）的分泌、酸性pH和 消化酶这一阶段将是适得其反的颗粒的完整性，我们的目的是 淋巴结内递送。此外，从吞噬细胞释放颗粒不是固有性质。但是，我想， 这种行为是有先例的。真菌细胞，新型隐球菌，eleclampicvomocytosis或（非溶解性 胞吐作用）。人们对这一现象的机制仍知之甚少 现象。一些报告指出，在这种活动期间，吞噬体的pH值增加， 表明Ca 2+通量可能是囊泡胞吐的关键。我们认为，阐明内- 推进这种活性的吞噬体物理化学条件可能对模拟物的设计有指导意义 微粒。此外，研究了在吞噬作用期间和 胞吐作用可以为我们提供线索，让我们了解吞噬体中哪些生物物理因素发生了改变。最终，我们长期的- 长期目标是开发可普遍部署的微粒疫苗平台，作为一种有效、持久 预防传染性病原体。该平台系统的研究和开发具有一定的潜力 来显著改变对大量免疫疾病的治疗。因此，拟议方案是 与国家普通医学科学研究所（NIGMS）的使命高度相关， 支持增加对生物过程的理解并为进步奠定基础的研究 在疾病诊断、治疗和预防方面。

DECODING VOMOCYTOSIS FOR CELL-MEDIATED, INTRA-LYMPH NODAL DELIVERY OF MICROPARTICLE VACCINES ABSTRACT/ SUMMARY The Immuno-modulatory Biomaterials Laboratory at UC Davis focuses on the development of novel biomaterial systems that can manipulate the immune system. Our goal is to design the next generation of immunotherapeutics for applications in immune-related diseases. This multidisciplinary work incorporates aspects of biomaterials engineering, drug delivery, immunology, biochemistry, and cell physiology. One of the main research programs in our lab is understanding controlled vomocytosis (non-lytic exocytosis) of particulate matter from phagocytic cells. Our primary motivation for elucidation of this process is the development of a `smart' microparticle system that can accomplish phagocytic cell-mediated delivery of vaccines directly to the lymph node-residing cells. Phagocytic cells (particularly macrophages [MΦ] and dendritic cells [DCs])) have evolved to circulate through peripheral tissue, engulf foreign particulate matter and traffick to the lymph node following uptake to relay information about the peripheral environment to cellular agents of the adaptive immune system. Evidently, phagocytic cells can transport particulate materials from peripheral tissues to lymphatic organs. Typically, following phagocytosis, and as phagocytic cells traverse the lymphatic vessels, materials that have been engulfed are taken into the phagosome (vacuole in the cytoplasm of a cell containing a phagocytosed particles) where a degradative process occurs due to secretion of reactive oxygen species (ROS), acidic pH and digestive enzymes. This stage would be counterproductive to the integrity of a particulate, and our purpose of intra-lymph nodal delivery. Moreover, release of particles from the phagocytic cell is not an intrinsic quality. But, there is precedent for such behavior. The fungal cell, Cryptococcus neoformans, elicits vomocytosis or (non-lytic exocytosis) from macrophages. There is still little understanding about the mechanisms governing this phenomena. Some reports indicate that there is an increase in pH of the phagosome during this activity, others have suggested Ca2+ flux could be pivotal for vesicle exocytosis. We believe that elucidation of the intra- phagosomal physico-chemical conditions that propel this activity could be instructive for the design of a mimetic microparticle. Further, investigation of the transcriptomal signatures between the period of phagocytosis and exocytosis could give us clues as to what biophysical factors are altered in the phagosome. Ultimately, our long- term goal is develop universally-deployable, microparticle vaccine platform as an effective, long-lasting prophylactic against infectious agents. The research and development of this platform system has the potential to significantly transform the treatment of a plethora of immune pathologies. Therefore, the proposed program is highly relevant to the mission of the National Institute of General Medical Sciences (NIGMS), which pertains to supporting research that increases understanding of biological processes and lays the foundation for advances in disease diagnosis, treatment and prevention.