Document Detail


Quantitative isolation of mouse and human intestinal intraepithelial lymphocytes by elutriation centrifugation.
MedLine Citation:
PMID:  19278662     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Intestinal intraepithelial lymphocytes (IEL) are specialized subsets of T cells with distinct functional capacities. While some IEL subsets are circulating, others such as CD8alphaalpha TCRalphabeta IEL are believed to represent non-circulating resident T cell subsets [Sim, G.K., Intraepithelial lymphocytes and the immune system. Adv. Immunol., 1995. 58: 297-343.]. Current methods to obtain enriched preparations of intraepithelial lymphocytes are mostly based on Percoll density gradient or magnetic bead-based technologies [Lundqvist, C., et al., Isolation of functionally active intraepithelial lymphocytes and enterocytes from human small and large intestine. J. Immunol. Methods, 1992. 152(2): 253-263.]. However, these techniques are hampered by a generally low yield of isolated cells, and potential artifacts due to the interference of the isolation procedure with subsequent functional assays, in particular, when antibodies against cell surface markers are required. Here we describe a new method for obtaining relatively pure populations of intestinal IEL (55-75%) at a high yield (>85%) by elutriation centrifugation. This technique is equally suited for the isolation and enrichment of intraepithelial lymphocytes of both mouse and human origin. Time requirements for fractionating cell suspensions by elutriation centrifugation are comparable to Percoll-, or MACS-based isolation procedures. Hence, the substantially higher yield and the consistent robust enrichment for intraepithelial lymphocytes, together with the gentle treatment of the cells during elutriation that does not interfere with subsequent functional assays, are important aspects that are in favor of using this elegant technology to obtain unmanipulated, unbiased populations of intestinal intraepithelial lymphocytes, and, if desired, also of pure epithelial cells.
Authors:
Elisa Binda; Dominik Erhart; Mirjam Schenk; Christel Zufferey; Pietro Renzulli; Christoph Mueller
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-03-09
Journal Detail:
Title:  Journal of immunological methods     Volume:  344     ISSN:  1872-7905     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-04-29     Completed Date:  2009-05-15     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  26-34     Citation Subset:  IM    
Affiliation:
Institute of Pathology, University of Bern, Bern, Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Separation / methods*
Cell Survival
Centrifugation / methods*
Humans
Intestinal Mucosa / immunology*
Lymphocyte Activation
Lymphocytes / immunology*
Mice
Mice, Inbred C57BL

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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