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[Abstract] [Full Text] [PDF]
In vivo and in vitro techniques for comparative study of antiviral T-cell responses in the amphibian Xenopus Heidi Morales1 and Jacques Robert1* 1 Department of Microbiology and Immunology, University of Rochester Medical Center. Rochester, NY 14642. Biol. Proced. Online 2008;10:1-8. doi:10.1251/bpo137 Indexing terms: xenopus laevis; ranavirus; T-Lymphocytes. Abbreviations: Ag, antigen; APBS, amphibian phosphate buffered saline; BrdU, bromodeoxyuridine; CFSE, carboxyfluorescein diacetate succinimidyl ester; FACS, fluorescence activated cell sorting; mAb, monoclonal antibody; MHC, major histocompatibility complex. |
Activation of lymphocytes in mammals is often quantified by measuring the amount of proliferation during the expansion phase of an immune response. Bromodeoxyuridine (BrdU) incorporation and carboxyfluorescein diacetate succinimidyl ester (CFSE) dilution assays are some of the techniques widely used in mammalian studies of pathogen-induced proliferation and provide a convenient way of quantifying the cellular response. We have extended the use of these proliferation assays to the amphibian Xenopus laevis. We have developed this species as a valuable comparative model to study immunity against a well-known amphibian pathogen, Frog Virus 3 (FV3). Fluorescence activated cell sorting was used to assess the level of BrdU incorporation of lymphocytes in vivo and CFSE dilution in an in vitro activation assay. Both techniques have shown that splenic lymphocytes proliferate specifically upon FV3 challenge. This indicates that common methods for detection of proliferation upon immunologic challenge are easily applied to other vertebrate species, as it highlights the evolutionary conservation of the proliferative nature of immune responses throughout vertebrate phyla. |