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Interrogating Adaptive Immunity Using LCMV.

Tanushree Dangi1, Young Rock Chung1, Nicole Palacio1

  • 1Microbiology-Immunology, Northwestern University Feinberg School of Medicine, Chicago, Illinois.

Current Protocols in Immunology
|September 17, 2020
PubMed
Summary

This article details methods for studying lymphocytic choriomeningitis virus (LCMV) infection in mice. It covers LCMV infection models, immune response quantification, and transgenic systems for research.

Keywords:
LCMV modelacute viral infectionchronic viral infectionimmune exhaustionimmune memory

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Area of Science:

  • Virology
  • Immunology
  • Infectious Diseases

Background:

  • The lymphocytic choriomeningitis virus (LCMV) is a widely used model system for studying viral infections and immune responses.
  • Previous contributions have established foundational knowledge, necessitating an updated overview of current methodologies.
  • Understanding LCMV infection dynamics is crucial for developing effective antiviral strategies and vaccines.

Purpose of the Study:

  • To provide a comprehensive update on technical aspects and protocols for studying the lymphocytic choriomeningitis virus (LCMV) system.
  • To guide researchers in selecting appropriate LCMV infection models, routes, and viral strains.
  • To detail methods for quantifying virus-specific immune responses and utilizing transgenic systems in LCMV research.

Main Methods:

  • LCMV infection protocols in mice, including route selection and viral stock preparation.
  • Assays for measuring LCMV titers, such as plaque assays and immunofluorescence focus assays (IFA).
  • Methods for assessing T cell and B cell responses, including tetramer staining, intracellular cytokine staining (ICS), antibody-secreting cell enumeration, limiting dilution assays (LDA), and ELISA.
  • Protocols for utilizing transgenic models, such as P14 cell transfer and comparative T cell expansion studies.

Main Results:

  • Detailed protocols are provided for various aspects of LCMV research, from infection to immune response measurement.
  • Specific methods are outlined for quantifying viral load, CD8 T cell responses, T cell cytokine production, and B cell antibody secretion.
  • The utility of transgenic systems in dissecting specific immune cell functions during LCMV infection is demonstrated.

Conclusions:

  • This article serves as an updated resource for researchers utilizing the LCMV model system.
  • The comprehensive protocols facilitate robust investigation into viral pathogenesis and host immune responses.
  • The described methods and transgenic systems enable detailed analysis of cellular and humoral immunity against LCMV.