Interrogating Adaptive Immunity Using LCMV

Abstract

In this invited article, we explain technical aspects of the lymphocytic choriomeningitis virus (LCMV) system, providing an update of a prior contribution by Matthias von Herrath and J. Lindsay Whitton. We provide an explanation of the LCMV infection models, highlighting the importance of selecting an appropriate route and viral strain. We also describe how to quantify virus-specific immune responses, followed by an explanation of useful transgenic systems. Specifically, our article will focus on the following protocols. © 2020 Wiley Periodicals LLC.

Basic Protocol 1: LCMV infection routes in mice

Support Protocol 1: Preparation of LCMV stocks

ASSAYS TO MEASURE LCMV TITERS

Support Protocol 2: Plaque assay

Support Protocol 3: Immunofluorescence focus assay (IFA) to measure LCMV titer

MEASUREMENT OF T CELL AND B CELL RESPONSES TO LCMV INFECTION

Basic Protocol 2: Triple tetramer staining for detection of LCMV-specific CD8 T cells

Basic Protocol 3: Intracellular cytokine staining (ICS) for detection of LCMV-specific T cells

Basic Protocol 4: Enumeration of direct ex vivo LCMV-specific antibody-secreting cells (ASC)

Basic Protocol 5: Limiting dilution assay (LDA) for detection of LCMV-specific memory B cells

Basic Protocol 6: ELISA for quantification of LCMV-specific IgG antibody

Support Protocol 4: Preparation of splenic lymphocytes

Support Protocol 5: Making BHK21-LCMV lysate

Basic Protocol 7: Challenge models

TRANSGENIC MODELS

Basic Protocol 8: Transfer of P14 cells to interrogate the role of IFN-I on CD8 T cell responses

Basic Protocol 9: Comparing the expansion of naïve versus memory CD4 T cells following chronic viral challenge