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.
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Abstract Basic Protocol 1 : LCMV infection routes in miceSupport Protocol 1 : Preparation of LCMV stocksASSAYS TO MEASURE LCMV TITERS Support Protocol 2 : Plaque assaySupport Protocol 3 : Immunofluorescence focus assay (IFA) to measure LCMV titerMEASUREMENT OF T CELL AND B CELL RESPONSES TO LCMV INFECTION Basic Protocol 2 : Triple tetramer staining for detection of LCMV‐specific CD8 T cellsBasic Protocol 3 : Intracellular cytokine staining (ICS) for detection of LCMV‐specific T cellsBasic 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 cellsBasic Protocol 6 : ELISA for quantification of LCMV‐specific IgG antibodySupport Protocol 4 : Preparation of splenic lymphocytesSupport Protocol 5 : Making BHK21‐LCMV lysateBasic Protocol 7 : Challenge modelsTRANSGENIC MODELS Basic Protocol 8 : Transfer of P14 cells to interrogate the role of IFN‐I on CD8 T cell responsesBasic Protocol 9 : Comparing the expansion of naïve versus memory CD4 T cells following chronic viral challenge -
Palacio, Nicole ; Dangi, Tanushree ; Chung, Young Rock ; Wang, Yidan ; Loredo-Varela, Juan Luis ; Zhang, Zhongyao ; Penaloza-MacMaster, Pablo ( , Journal of Experimental Medicine)
Type I interferons (IFN-I) are a major antiviral defense and are critical for the activation of the adaptive immune system. However, early viral clearance by IFN-I could limit antigen availability, which could in turn impinge upon the priming of the adaptive immune system. In this study, we hypothesized that transient IFN-I blockade could increase antigen presentation after acute viral infection. To test this hypothesis, we infected mice with viruses coadministered with a single dose of IFN-I receptor–blocking antibody to induce a short-term blockade of the IFN-I pathway. This resulted in a transient “spike” in antigen levels, followed by rapid antigen clearance. Interestingly, short-term IFN-I blockade after coronavirus, flavivirus, rhabdovirus, or arenavirus infection induced a long-lasting enhancement of immunological memory that conferred improved protection upon subsequent reinfections. Short-term IFN-I blockade also improved the efficacy of viral vaccines. These findings demonstrate a novel mechanism by which IFN-I regulate immunological memory and provide insights for rational vaccine design.