Interrogating Adaptive Immunity Using LCMV. (6th July 2020)

• Record Type: Journal Article
• Authors: Dangi, Tanushree; Chung, Young Rock; Palacio, Nicole; Penaloza‐MacMaster, Pablo
• Is Part Of: Current protocols in immunology. Volume 130(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-07-06
• Subjects: acute viral infection -- chronic viral infection -- immune exhaustion -- immune memory -- LCMV model; Immunology -- Laboratory manuals; Medical protocols; Immunologic Techniques; Medical protocols; Immunology; Laboratory manuals; Reference works; 571.96028
• DOI: 10.1002/cpim.99
• Languages: English
• ISSNs: 1934-3671

Abstract

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