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Publisher
Humana Press, New York, NY
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Authors: Stasya Zarling Urszula Krzych
Publish Date: 2015
Volume: , Issue: , Pages: 39-48
Abstract
Murine models of malaria such as Plasmodium berghei Pb and Plasmodium yoelii Py have been used for decades to identify correlates of protection associated with immunization using radiationattenuated sporozoites RAS To date RAS is the only known immunization regimen to consistently deliver 100 sterilizing immunity and is considered the “gold standard” of protection against malaria The ability to isolate lymphocytes directly from the liver of immune mice has facilitated the identification of correlates of protection at the site of infection Liver CD8 T cells have been identified as a key factor in mediating protection against challenge with infectious Plasmodium sporozoites Liver CD3 + CD8 T cells can further be divided into subsets based on the expression of specific surface molecules and the increase of CD8 effector memory TEM cells identified by the phenotype CD44+CD62L− has been shown to mediate protection by releasing of IFNγ while CD8 central memory TCM cells CD44+CD62L+ are important for maintaining longterm protectionIdentification of multiple CD8 T cell subsets present in the liver relies on the ability to detect multiple surface markers simultaneously Polychromatic flow cytometry affords the user with the ability to distinguish multiple lymphocyte populations as well as subsets defined within each population In this chapter we present a basic 9color surface staining panel that can be used to identify CD8 TEM CD8 TCM shortlived effector cells SLECs and memory precursor cells MPECs as well as identify those cells which have recently undergone degranulation surface expression of CD107a This panel has been designed to allow for the addition of intracellular staining for IFNγ on other available channels such as PE as is discussed in another chapter for analysis of functional CD8 T cell responsesThe research described in this chapter was supported by the US Army Medical Research and Materiel Command The views of the authors do not purport to reflect the position of the Department of the Army or the Department of Defense Described procedures must be conducted under an IACUCapproved protocol in accordance with the Animal Welfare Act and the Guide for the Care and Use of Laboratory Animals NRC 2011
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