Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2088595 | Journal of Immunological Methods | 2010 | 5 Pages |
Abstract
Currently human T-cell responses to unpurified tissue extracts cannot be easily measured because current cell lysis methods yield a lysate that is toxic. Here we describe the optimization of a protocol for extracting proteins from human tissues in a format that is compatible with the functional analysis of human T cells. The tissue was homogenized in a mixture of butan-1-ol, acetonitrile and water and then lyophilized. Lyophilized protein extracts were dissolved in 8Â M urea because urea did not affect T-cell function when present at 0.08Â M or less. Using this method cytokine production and proliferation responses were detected from islet, acinar and spleen extracts. Hence, our method allows the rapid preparation of human tissue lysates in a format that is compatible with the analysis T-cell responses. We suggest that this method will facilitate the analysis of adaptive immune responses to tissues in transplantation, cancer and autoimmunity.
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Authors
Hayley C. Moon, Max Joffe, Helen E. Thomas, Thomas W.H. Kay, Stuart I. Mannering,