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Choosing and preparing antigen-presenting cells.

Clifford V Harding1

  • 1Case Western Reserve University, Cleveland, Ohio.

Current Protocols in Immunology
|April 25, 2008
PubMed
Summary
This summary is machine-generated.

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This study details methods for preparing activated macrophages and B lymphoblasts for antigen-presenting cell (APC) experiments. It offers protocols for eliciting macrophages using Listeria monocytogenes, ensuring high activation levels for research.

Area of Science:

  • Immunology and Cell Biology
  • Experimental Methods in Immunology

Background:

  • Selecting and obtaining suitable antigen-presenting cells (APCs) is a critical initial step in antigen-processing experiments.
  • Macrophages possess specific characteristics making them valuable for certain antigen-processing studies.

Purpose of the Study:

  • To describe reliable methods for preparing activated macrophages and other antigen-presenting cells for experimental use.
  • To provide detailed protocols for generating high-quality APCs essential for antigen-processing research.

Main Methods:

  • Eliciting peritoneal macrophages from mice via infection with Listeria monocytogenes for consistent, high-level activation.
  • Alternative macrophage isolation methods including sequential injection of L. monocytogenes and proteose peptone, concanavalin A (Con A) induction, or bone marrow isolation.

Related Experiment Videos

  • Preparation of L. monocytogenes stocks and L929-conditioned medium for culturing bone marrow-derived macrophages.

    • Main Results:

    • The Listeria monocytogenes infection method yields consistently activated macrophages, preferred for antigen-processing experiments.
    • Established protocols for various macrophage activation and isolation techniques, alongside B lymphoblast preparation.

    Conclusions:

    • Provides essential, practical protocols for researchers to obtain and activate macrophages and B lymphoblasts for antigen-presenting cell studies.
    • Facilitates reproducible antigen-processing experiments through standardized APC preparation methods.