University of Melbourne, St. Vincent's Hospital

2 ARTICLES PUBLISHED IN JoVE

Biology

Extraction of Tissue Antigens for Functional Assays

Andra Necula¹, Rochna Chand¹, Batool Albatat¹, 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.

Immunology and Infection

Quantification of Proliferating Human Antigen-specific CD4⁺ T Cells using Carboxyfluorescein Succinimidyl Ester

Anthony R. Di Carluccio¹, Eleonora Tresoldi¹, Michelle So¹, 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.