In vivo and in vitro techniques for comparative study of antiviral T-cell responses in the amphibian Xenopus

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Author: Morales, H
Author: Robert, J
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Methods Article

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.

* To whom correspondence should be addressed: Jacques Robert, Department of Microbiology and Immunology, University of Rochester Medical Center. Rochester, NY 14642. USA. Phone: (585) 275 1722. Fax: (716) 473 9573. Email: jacques_robert@urmc.rrochester.edu

Biol. Proced. Online 2008;10:1-8. doi:10.1251/bpo137
Submitted: September 06, 2007; Accepted: September 26, 2007; Published: January 17, 2008.

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.

Figure 1 Enlarged

Fig. 1:

Detection of Xenopus splenocyte proliferation 6 days post-FV3 infection by BrdU assay. Uninfected control frogs and frogs infected for 6 days with FV3 were treated with BrdU 48 hrs before the assay. Cells were then permeabilized, treated with DNase, stained with FITC-conjugated anti-BrdU mAb, and analyzed by FACS (Representative of six different experiments). Ten thousand total events were gated based on side and forward scatter profiles (A, C). The arrows depict cells with higher forward scatter that increase in number in the infected sample. The percent of gated BrdU positive cells is shown in the histograms (B, D).

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