A method for extracting tissue proteins for use in lymphocyte function assays

J Immunol Methods. 2010 Jul 31;359(1-2):56-60. doi: 10.1016/j.jim.2010.05.002. Epub 2010 May 12.


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 8M urea because urea did not affect T-cell function when present at 0.08 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Proliferation
  • Chemical Fractionation / methods*
  • Cytokines / biosynthesis
  • Cytokines / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoassay / methods
  • Interferon-gamma / chemistry
  • Interferon-gamma / immunology
  • Islets of Langerhans / chemistry*
  • Islets of Langerhans / cytology
  • Islets of Langerhans / immunology
  • Lymphocyte Activation* / drug effects
  • Proteins / chemistry
  • Proteins / isolation & purification*
  • Proteins / pharmacology
  • Spleen / chemistry*
  • Spleen / cytology
  • Spleen / immunology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*
  • Tissue Extracts / chemistry*


  • Cytokines
  • Proteins
  • Tissue Extracts
  • Interferon-gamma