Abstract
The isolation of intestinal intraepithelial lymphocytes (IEL) is a major prerequisite for the investigation of cellular and molecular cross-talk in the intestinal mucosa. Since intestinal epithelial cells exhibit distinct functional features at the villi tip and crypt levels, such differences could extend to IEL. We developed a mechanical procedure for isolation of IEL from these distinct epithelial sites to test our hypothesis. Cells isolated from the intestinal epithelium by sequential incubations under stirring were segregated based upon their alkaline phosphatase (AP) activity since villi tip and crypt fractions expressed high and low AP activity, respectively. IEL preparations obtained after a further purification step in Percoll gradient contained > 90% Integrin alpha IEL chain+, CD3+ T cells, and no Ig+ cells. Villi tip IEL preparations possessed increased numbers of low density IEL when compared to crypt IEL, suggesting that distinct IEL-epithelial cell interactions occur at the intestinal villi tip and crypt levels.
References
Jul 1, 1992·The Journal of Experimental Medicine·P PoussierM Julius
Feb 1, 1991·The Journal of Experimental Medicine·D Guy-GrandP Vassalli
Jul 1, 1990·The Journal of Experimental Medicine·A BandeiraA Coutinho
Dec 1, 1990·European Journal of Immunology·J L Viney, T T MacDonald
Dec 31, 1990·Biochemical and Biophysical Research Communications·P G Traber
Dec 1, 1988·Nature·M BonnevilleS Tonegawa
Dec 18, 1989·FEBS Letters·O NorénG H Hansen
Jun 30, 1988·Nature·T Goodman, L Lefrançois
Jun 1, 1985·Journal of Cellular Physiology·M RoussetA Zweibaum
Dec 1, 1995·The Journal of Experimental Medicine·G R KlimpelJ W Peterson
Dec 1, 1994·Immunity·L PuddingtonL Lefrançois
Jun 20, 1995·Proceedings of the National Academy of Sciences of the United States of America·H KomanoS Itohara
Jun 1, 1994·European Journal of Immunology·M YamamotoH Kiyono
Oct 1, 1993·European Journal of Immunology·R C FindlyA C Hayday
Apr 16, 1996·Proceedings of the National Academy of Sciences of the United States of America·K FujihashiH Kiyono
Citations
Oct 15, 2008·Journal of Pediatric Gastroenterology and Nutrition·Jody L GookinMaria R Stone
Feb 17, 2000·The Journal of Clinical Investigation·A P NarenK L Kirk
Aug 8, 2009·Bioscience, Biotechnology, and Biochemistry·Jae-Sung LeeSatoshi Ishizuka
Sep 30, 1998·Gastroenterology·S KawabataH Kiyono
Jul 16, 2003·Laboratory Investigation; a Journal of Technical Methods and Pathology·Hiroshi TamagawaHiroshi Kiyono