GSBS Student Publications

Title

A new isolation method for rat intraepithelial lymphocytes

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Medicine

Date

6-5-1999

Document Type

Article

Medical Subject Headings

*ADP Ribose Transferases; Animals; Antigens, CD4; Antigens, CD45; Antigens, CD8; Antigens, Differentiation, T-Lymphocyte; Centrifugation, Density Gradient; Epithelial Cells; Flow Cytometry; Histocompatibility Antigens; Immunophenotyping; Killer Cells, Natural; Lymphocyte Subsets; Lymphocytes; *Membrane Glycoproteins; Peyer's Patches; Povidone; Rats; Rats, Inbred Strains; Receptors, Antigen, T-Cell, alpha-beta; Receptors, Antigen, T-Cell, gamma-delta; Silicon Dioxide

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Intraepithelial lymphocytes (IELs) play critical roles in gut immunity. In mice, gammadelta T cells are a large component of the IEL population. In the rat, gammadelta IELs are reportedly much less common, but technical issues suggest that previous analyses should be interpreted cautiously. The study of IELs in rats has been impeded by isolation procedures that are lengthy and complex, leading to small cell yields. For this reason, it is possible that rat IELs analyzed in previous studies have not been representative of the entire IEL compartment. We report a new method for the isolation of rat IELs that is based on the selective removal of intestinal epithelial cells under conditions that leave the basement membrane undisturbed. The method is rapid and requires neither enzymatic digestion, nor surgical removal of Peyer's patches, nor vigorous mechanical manipulation of the intestine. The yield of rat IELs using this method is 5- to 10-fold greater than that reported for other methods. Morphological and phenotypic analyses demonstrated that the purified cell population is comprised of IELs and is not contaminated with lamina propria or Peyer's patch lymphocytes. Phenotypic analysis revealed five major subsets of IELs based on differential cell surface expression of CD4, CD8, and alphabeta T cell receptor (TcR). Among the alphabetaTcR- cells was a population of gammadelta T cells present at levels not previously detected. The isolation of IEL sub-populations using this methodology should facilitate studies of the function of these cells in gut immunity.

Rights and Permissions

Citation: J Immunol Methods. 1999 Apr 22;224(1-2):111-27.

Related Resources

Link to Article in PubMed

Journal Title

Journal of immunological methods

PubMed ID

10357212