Interrogating adaptive immunity using LCMV
Current protocols in immunology, 2020•Wiley Online Library
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 …
(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 …
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
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