包膜乳胶珠作为人工细胞用于受体/配体相互作用的定量研究

Q2 Immunology and Microbiology Current Protocols in Immunology Pub Date : 2020-11-04 DOI:10.1002/cpim.111
Doris Urlaub, Carsten Watzl
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引用次数: 2

摘要

在免疫应答过程中,细胞间的相互作用往往是调节免疫细胞活动所必需的。为了了解这一过程的细节,有必要以定量的方式研究单个受体/配体的相互作用。然而,当使用真实细胞进行刺激时,这通常是非常困难甚至不可能的。在这里,我们提出了一种使用细胞大小的乳胶珠进行此类研究的方法。这些小珠可以以确定的和可量化的方式涂覆激动性抗体或特定配体。这创造了在三维系统中滴定特定受体的刺激强度的可能性。以自然杀伤(NK)细胞为例,我们展示了这些珠子如何用于刺激NK细胞反应。©2020作者。基本方案1:用抗体对乳胶珠进行共价包衣基本方案2:用QIFIKIT®对乳胶珠进行共价包衣以结合生物素化蛋白替代方案1:用QIFIKIT®支持方案对乳胶珠进行蛋白量的定量:在自然杀伤细胞脱粒试验中对乳胶珠进行功能测试
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Coated Latex Beads as Artificial Cells for Quantitative Investigations of Receptor/Ligand Interactions

Cellular interactions are often essential to regulate immune cell activities during an immune response. To understand the details of this process, it is necessary to study individual receptor/ligand interactions in a quantitative fashion. However, this is often very difficult or even impossible when using real cells for stimulation. Here, we present a method to use cell-sized latex beads for such studies. These beads can be coated with agonistic antibodies or specific ligands in a defined and quantifiable fashion. This creates the possibility of titrating the strength of the stimulation for a specific receptor in a three-dimensional system. Using natural killer (NK) cells as an example, we demonstrate how these beads can be used to stimulate NK cell responses. © 2020 The Authors.

Basic Protocol 1: Covalent coating of latex beads with antibodies

Basic Protocol 2: Quantification of the amount of antibodies on the beads with the QIFIKIT®

Alternate Protocol 1: Covalent coating of latex beads with streptavidin to bind biotinylated proteins

Alternate Protocol 2: Quantification of the amount of protein on the beads with the QIFIKIT®

Support Protocol: Functional testing of the beads in a natural killer cell degranulation assay

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Current Protocols in Immunology
Current Protocols in Immunology Immunology and Microbiology-Immunology
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