title: Visualizing chemokine-dependent T cell activation and migration in response to central nervous system infection.
abstract: In response to central nervous system (CNS) injury and infection, astrocytes, neurons, and CNS vasculature express several chemokines, including CCL21. Quantitative polymerase chain reaction (qPCR), western blot, and immunohistochemical methods can quantify mRNA and protein expression. However, these methods do not quantify chemokine bioavailability and bioactivity, variables modified by many environmental factors including composition of the extracellular matrix (ECM). Here we illustrate how two-photon microscopy and carboxyfluorescein succinimidyl ester (CFSE or CFDA SE) labeling of T cells coupled with flow cytometry can be used as tools to assess chemokine-mediated regulation of T cell proliferation, activation, and migration.
authors.affiliation: Division of Biomedical Sciences, Center for Glial-Neuronal Interactions, University of California Riverside, Riverside, CA, USA.
authors.forename: Monica J
authors.initials: MJ
authors.lastname: Carson
authors.forename: Emma H
authors.initials: EH
authors.lastname: Wilson
mesh_terms: D000818:Animals; D001921:Brain / Q000276:immunology / Q000378:metabolism*; D049109:Cell Proliferation; D058948:Cell Tracking / Q000379:methods; D002494:Central Nervous System Infections / Q000276:immunology / Q000378:metabolism*; D054421:Chemokine CCL21 / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism*; D002633:Chemotaxis; D004195:Disease Models, Animal; D005434:Flow Cytometry; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D015398:Signal Transduction; D013388:Succinimides / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000637:transplantation; D013997:Time Factors
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D054421:Chemokine CCL21; D018925:Chemokines; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides