• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

使用免疫荧光技术在巨噬细胞中快速评估抗体介导保护作用的实验方案。

Protocol for rapid evaluation of antibody-mediated protection in macrophages using immunofluorescence techniques.

作者信息

Zeng Lingyuan, Li Hao

机构信息

National Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.

National Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103324. doi: 10.1016/j.xpro.2024.103324. Epub 2024 Sep 19.

DOI:10.1016/j.xpro.2024.103324
PMID:39302838
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11437938/
Abstract

Antibodies have been shown to provide protection against tuberculosis (TB). It is important to identify the qualitative and quantitative protective effects of antibodies before a clinical study. Here, we present a protocol to evaluate antibody-mediated protection in macrophages using confocal fluorescence microscopy and flow cytometry. We describe steps for bacteria and macrophage preparation, cell infection, phagocytosis, and phagosome maturation analysis. For complete details on the use and execution of this protocol, please refer to Zeng et al..

摘要

抗体已被证明可提供抗结核病（TB）的保护作用。在进行临床研究之前，确定抗体的定性和定量保护作用很重要。在这里，我们提出了一种使用共聚焦荧光显微镜和流式细胞术评估巨噬细胞中抗体介导的保护作用的方案。我们描述了细菌和巨噬细胞制备、细胞感染、吞噬作用和吞噬体成熟分析的步骤。有关此方案的使用和执行的完整详细信息，请参考曾等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/3adc3132963f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/1c6bf2a61fd8/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/abf3debfdee4/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/5a2d06db21a1/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/f0bf0a0032c3/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/b4bb9f373eb2/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/3adc3132963f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/1c6bf2a61fd8/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/abf3debfdee4/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/5a2d06db21a1/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/f0bf0a0032c3/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/b4bb9f373eb2/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8388/11437938/3adc3132963f/gr5.jpg

相似文献

1
Protocol for rapid evaluation of antibody-mediated protection in macrophages using immunofluorescence techniques.使用免疫荧光技术在巨噬细胞中快速评估抗体介导保护作用的实验方案。
STAR Protoc. 2024 Dec 20;5(4):103324. doi: 10.1016/j.xpro.2024.103324. Epub 2024 Sep 19.
2
Protocol to characterize mouse dural mast cells by flow cytometry and immunofluorescence.通过流式细胞术和免疫荧光对小鼠硬脑膜肥大细胞进行表征的方案。
STAR Protoc. 2024 Dec 20;5(4):103364. doi: 10.1016/j.xpro.2024.103364. Epub 2024 Oct 7.
3
Protocol for detecting macrophage-mediated cancer cell phagocytosis in vitro and in vivo.体外和体内检测巨噬细胞介导的癌细胞吞噬作用的方案。
STAR Protoc. 2023 Mar 17;4(1):101940. doi: 10.1016/j.xpro.2022.101940. Epub 2022 Dec 14.
4
Quantitative Immunofluorescence to Study Phagosome Maturation.用于研究吞噬体成熟的定量免疫荧光法。
Methods Mol Biol. 2017;1519:113-123. doi: 10.1007/978-1-4939-6581-6_8.
5
Phagocyte interactions with Mycobacterium tuberculosis--Simultaneous analysis of phagocytosis, phagosome maturation and intracellular replication by imaging flow cytometry.吞噬细胞与结核分枝杆菌的相互作用——通过成像流式细胞术同时分析吞噬作用、吞噬体成熟和细胞内复制
J Immunol Methods. 2015 Dec;427:73-84. doi: 10.1016/j.jim.2015.10.003. Epub 2015 Oct 23.
6
Evaluation of mTORC1 signaling in mouse atherosclerotic macrophages by flow cytometry and immunofluorescence.通过流式细胞术和免疫荧光法评估小鼠动脉粥样硬化巨噬细胞中的 mTORC1 信号转导。
STAR Protoc. 2022 Dec 16;3(4):101665. doi: 10.1016/j.xpro.2022.101665. Epub 2022 Sep 11.
7
Click-chemistry-based protocol to quantitatively assess fatty acid uptake by Mycobacterium tuberculosis in axenic culture and inside mouse macrophages.基于点击化学的方案定量评估分枝杆菌在体外培养和小鼠巨噬细胞内的脂肪酸摄取。
STAR Protoc. 2023 Mar 17;4(1):102062. doi: 10.1016/j.xpro.2023.102062. Epub 2023 Jan 24.
8
Efferocytosis assay to quantify the engulfment and acidification of apoptotic cells by macrophages using flow cytometry.利用流式细胞术检测巨噬细胞吞噬和酸化凋亡细胞的效率分析实验。
STAR Protoc. 2024 Sep 20;5(3):103215. doi: 10.1016/j.xpro.2024.103215. Epub 2024 Jul 27.
9
Detection of LC3-Associated Phagocytosis (LAP).LC3 相关噬菌作用（LAP）的检测。
Curr Protoc Cell Biol. 2020 Jun;87(1):e104. doi: 10.1002/cpcb.104.
10
Isolation and characterization of a protective monoclonal antibody targeting outer membrane protein (OmpA) against tuberculosis.一种针对结核分枝杆菌外膜蛋白（OmpA）的保护性单克隆抗体的分离与鉴定
Microbiol Spectr. 2025 Apr;13(4):e0294224. doi: 10.1128/spectrum.02942-24. Epub 2025 Feb 18.

引用本文的文献

1
Isolation and characterization of a protective monoclonal antibody targeting outer membrane protein (OmpA) against tuberculosis.一种针对结核分枝杆菌外膜蛋白（OmpA）的保护性单克隆抗体的分离与鉴定
Microbiol Spectr. 2025 Apr;13(4):e0294224. doi: 10.1128/spectrum.02942-24. Epub 2025 Feb 18.
2
A subunit vaccine Ag85A-LpqH focusing on humoral immunity provides substantial protection against tuberculosis in mice.一种聚焦于体液免疫的亚单位疫苗Ag85A-LpqH为小鼠提供了对结核病的显著保护。
iScience. 2024 Dec 20;28(1):111568. doi: 10.1016/j.isci.2024.111568. eCollection 2025 Jan 17.

本文引用的文献

1
Immunogenicity and protective efficacy of Ag85A and truncation of PstS1 fusion protein vaccines against tuberculosis.Ag85A与截短的PstS1融合蛋白疫苗对结核病的免疫原性及保护效力
Heliyon. 2024 Feb 23;10(5):e27034. doi: 10.1016/j.heliyon.2024.e27034. eCollection 2024 Mar 15.
2
An anti-LpqH human monoclonal antibody from an asymptomatic individual mediates protection against Mycobacterium tuberculosis.一名无症状个体产生的抗LpqH人源单克隆抗体可介导对结核分枝杆菌的保护作用。
NPJ Vaccines. 2023 Aug 25;8(1):127. doi: 10.1038/s41541-023-00710-1.
3
Antibodies Targeting the Cell Wall Induce Protection against Virulent Mycobacterium bovis Infection.
靶向细胞壁的抗体可诱导对强毒力牛分枝杆菌感染的保护作用。
Microbiol Spectr. 2023 Feb 27;11(2):e0343122. doi: 10.1128/spectrum.03431-22.
4
BCG vaccination strategies against tuberculosis: updates and perspectives.BCG 疫苗接种策略防治结核病：更新与展望。
Hum Vaccin Immunother. 2021 Dec 2;17(12):5284-5295. doi: 10.1080/21645515.2021.2007711.
5
Human antibodies targeting a Mycobacterium transporter protein mediate protection against tuberculosis.靶向分枝杆菌转运蛋白的人源抗体可介导对结核病的保护。
Nat Commun. 2021 Jan 27;12(1):602. doi: 10.1038/s41467-021-20930-0.
6
Antibodies and tuberculosis: finally coming of age?抗体与结核病：终于迎来突破？
Nat Rev Immunol. 2018 Sep;18(9):591-596. doi: 10.1038/s41577-018-0028-0.
7
Latently and uninfected healthcare workers exposed to TB make protective antibodies against .潜伏感染和未感染的医护人员接触结核病后会产生针对……的保护性抗体。
Proc Natl Acad Sci U S A. 2017 May 9;114(19):5023-5028. doi: 10.1073/pnas.1611776114. Epub 2017 Apr 24.
8
Over-Expression of the Mycobacterial Trehalose-Phosphate Phosphatase OtsB2 Results in a Defect in Macrophage Phagocytosis Associated with Increased Mycobacterial-Macrophage Adhesion.分枝杆菌海藻糖磷酸磷酸酶OtsB2的过表达导致巨噬细胞吞噬缺陷，这与分枝杆菌-巨噬细胞粘附增加有关。
Front Microbiol. 2016 Nov 4;7:1754. doi: 10.3389/fmicb.2016.01754. eCollection 2016.