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St. Vincent's Institute of Medical Research

2 ARTICLES PUBLISHED IN JoVE

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Biology

Extraction of Tissue Antigens for Functional Assays
Andra Necula 1, Rochna Chand 1, Batool Albatat 1, Stuart I. Mannering 1,2
1Immunology and Diabetes Unit, St. Vincent's Institute of Medical Research, 2Department of Medicine, University of Melbourne

A simple protocol for preparing extracts of human tissue to be used as a source of antigens in functional T-cell assays is described. This method allows T-cell responses to tissue-derived antigens to be measured in vitro.

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Immunology and Infection

Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester
Anthony R. Di Carluccio 1, Eleonora Tresoldi 1, Michelle So 1, Stuart I. Mannering 1,2
1Immunology and Diabetes Unit, St. Vincent's Institute of Medical Research, 2Department of Medicine, University of Melbourne, St. Vincent's Hospital

Presented here is a protocol for measuring proliferating CD4+ T cells in response to antigenic proteins or peptides using dye dilution. This assay is particularly sensitive to rare antigen-specific T cells and can be modified to facilitate cloning of antigen-specific cells.

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