隔离和树突细胞吸收来自 Echinococcus granulosus 的小细胞外囊泡
María Celeste Nicolao1, Maia Chop2, Christian Rodriguez Rodrigues3
1IIPROSAM, Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata (UNMdP); Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).
Journal of visualized experiments : JoVE
|April 14, 2025
概括
这项研究详细介绍了一项协议,用于从 Echinococcus granulosus 中分离小细胞外囊泡 (sEV),并分析它们被树突细胞吸收. 这项研究有助于理解寄生虫宿主通信和免疫调节.
科学领域:
- 寄生虫学的寄生虫学
- 免疫学 免疫学 免疫学
- 细胞生物学 细胞生物学
背景情况:
- 虫分泌细胞外囊泡 (EVs),用于寄生虫与宿主之间的通信.
- 寄生虫的小细胞外囊泡 (sEV) 转移抗原,影响宿主免疫力和寄生虫的生存.
- 了解寄生sEVs和宿主免疫细胞之间的相互作用对于开发治疗策略至关重要.
研究的目的:
- 提供一个详细的协议来隔离和表征来自 Echinococcus granulosus 的 sEV.
- 分析由小鼠骨髓衍生的树突细胞 (BMDCs) 吸收寄生虫sEVs.
- 建立评估树状细胞成熟和暴露于寄生虫sEVs后抗原呈现的方法.
主要方法:
- 使用超离心法从 Echinococcus granulosus 幼虫培养物中分离和净化 sEV.
- 通过动态光散射和传输电子显微镜对sEVs的表征.
- 使用Flt3L隔离和体外分化小鼠BMDCs,随后暴露于寄生虫sEVs.
主要成果:
- 成功分离和定量了埃奇诺科克细粒菌sEVs.
- 证明树突细胞吸收寄生虫SEVs.
- 建议的方法 (聚焦显微镜,流细胞计) 评估树突细胞成熟和表型变化后sEV暴露.
结论:
- 描述的协议有助于研究寄生sEVs及其与宿主免疫细胞的相互作用.
- 这项研究为研究 cestode 免疫调节机制提供了基础.
- 该协议可适应各种研究应用,包括寄生虫培养和免疫细胞分析.
相关概念视频
Eukaryotic Compartmentalizations
One of the distinguishing features of eukaryotic cells is that they contain membrane-bound organelles, such as the nucleus and mitochondria, that carry out specialized functions. Since biological membranes are only selectively permeable to solutes, they help create a compartment with controlled conditions inside an organelle. These microenvironments are tailored to the organelle's specific functions and help isolate them from the surrounding cytosol.
For example, lysosomes in the animal cells...
For example, lysosomes in the animal cells...
Pinocytosis
Cells use energy-requiring bulk transport mechanisms to transfer large particles, or large amounts of small particles, into or out of the cell. The cells envelop the particles in spherical membranes called vesicles or vacuoles. Vesicles that transport material into the cell are built from the cell membrane. These vesicles encapsulate external molecules and transport them into the cell in a process called endocytosis.
Phagocytosis
Cells pull particles inward and engulf them in spherical vesicles in an energy-requiring process called endocytosis. Phagocytosis (“cellular eating”) is one of three major types of endocytosis. Cells use phagocytosis to take in large objects—such as other cells (or their debris), bacteria, and even viruses.The objective of phagocytosis is often destruction. Cells use phagocytosis to eliminate unwelcome visitors, like pathogens (e.g., viruses and bacteria). It is perhaps unsurprising, that many...
Endocytosis
Eukaryotic cells acquire nutrients for growth and proliferation. Nutrients and other molecules that require degradation are internalized from the extracellular space by a process called endocytosis. The term ‘endocytosis' was first coined by Christian de Duve in 1963.
Endocytosis always begins with the plasma membrane enclosing an incoming molecule to form a transport vesicle which, in some cases, can be coated with a protein called ‘clathrin.' Endocytosed material is either sorted through...
Endocytosis always begins with the plasma membrane enclosing an incoming molecule to form a transport vesicle which, in some cases, can be coated with a protein called ‘clathrin.' Endocytosed material is either sorted through...
Phagocytosis
Cells pull particles inward and engulf them in spherical vesicles in an energy-requiring process called endocytosis. Phagocytosis ("cellular eating") is one of three major types of endocytosis. Cells use phagocytosis to take in large objects, such as other cells (or their debris), bacteria, and even viruses.
The objective of phagocytosis is often destruction. Cells use phagocytosis to eliminate unwelcome visitors, like pathogens (e.g., viruses and bacteria). Many immune system cells, including...
The objective of phagocytosis is often destruction. Cells use phagocytosis to eliminate unwelcome visitors, like pathogens (e.g., viruses and bacteria). Many immune system cells, including...
Renewal of Intestinal Stem Cells
The intestinal epithelial lining rapidly renews every 4 to 5 days. The renewal is facilitated by intestinal stem cells (ISCs) located at the base of the crypt– a gland located at the bottom of each villus. ISCs divide asymmetrically to form new stem cells and progenitor daughter cells. The daughter cells are called transit-amplifying (TA) cells which move upwards along the crypt and either differentiate into absorptive cells– the enterocytes or secretory cells– including the goblet,...


