A new isolation method for rat intraepithelial lymphocytes
Identifieur interne : 001938 ( Main/Exploration ); précédent : 001937; suivant : 001939A new isolation method for rat intraepithelial lymphocytes
Auteurs : Derrick Todd [États-Unis] ; Amrik J. Singh [États-Unis] ; Dale L. Greiner [États-Unis] ; John P. Mordes [États-Unis] ; Aldo A. Rossini [États-Unis] ; Rita Bortell [États-Unis]Source :
- Journal of Immunological Methods [ 0022-1759 ] ; 1999.
English descriptors
- Teeft :
- Abtcr, Abtcrq, Abtcry, Additional phenotypic markers, Basement membrane, Bottom panel, Btcrq, Btcrq iels, Btcry, Cd4q, Cd4y, Cd8a, Cd8aq, Cd8ay, Cd8q, Cell number, Cell populations, Cell receptor, Clone, Comparable results, Cytometric, Differential expression, Epithelial, Epithelial cells, Epithelium, Fangmann, Flow cytometric analysis, Flow cytometry, Fluorescence intensity, Gdtcrq, Gdtcrq cells, Gdtcrq iels, Helgeland, High density, Horizontal axis, Iels, Immunol, Immunological, Immunological methods, Individual animals, Individual samples, Insets show fluorescence profiles, Intestinal, Intestinal epithelial cells, Intestine, Intraepithelial, Intraepithelial lymphocytes, Isolation procedure, Isotype control immunoglobulin, Kearsey, Lamina, Lamina propria, Large numbers, Lefrancois, Light microscopy, Lymph, Lymph node cells, Lymphocyte, Lymphoid, Lymphoid cells, Major subsets, Methodology, Middle panel, National institutes, Node, Nylon, Nylon wool, Nylon wool purification, Patch lymphocytes, Percoll, Percoll solution, Peripheral lymph node cells, Phenotype, Phenotypic, Phenotypic characteristics, Previous reports, Propria, Receptor, Representative profiles, Right panel, Rt6q cells, Same cell populations, Small intestine, Stadnyk, Subset, Takimoto, Teitelbaum, Third fluorochrome, Todd, Total iels, Unique staining patterns, Vertical axis, Waite.
Abstract
Abstract: Intraepithelial lymphocytes (IELs) play critical roles in gut immunity. In mice, γδ T cells are a large component of the IEL population. In the rat, γδ 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 αβ T cell receptor (TcR). Among the αβTcR− cells was a population of γδ 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.
Url:
DOI: 10.1016/S0022-1759(99)00015-0
Affiliations:
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Singh</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Greiner, Dale L" sort="Greiner, Dale L" uniqKey="Greiner D" first="Dale L" last="Greiner">Dale L. Greiner</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Mordes, John P" sort="Mordes, John P" uniqKey="Mordes J" first="John P" last="Mordes">John P. Mordes</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Rossini, Aldo A" sort="Rossini, Aldo A" uniqKey="Rossini A" first="Aldo A" last="Rossini">Aldo A. Rossini</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Bortell, Rita" sort="Bortell, Rita" uniqKey="Bortell R" first="Rita" last="Bortell">Rita Bortell</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Immunological Methods</title><title level="j" type="abbrev">JIM</title><idno type="ISSN">0022-1759</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1999">1999</date><biblScope unit="volume">224</biblScope><biblScope unit="issue">1–2</biblScope><biblScope unit="page" from="111">111</biblScope><biblScope unit="page" to="127">127</biblScope></imprint><idno type="ISSN">0022-1759</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0022-1759</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Abtcr</term><term>Abtcrq</term><term>Abtcry</term><term>Additional phenotypic markers</term><term>Basement membrane</term><term>Bottom panel</term><term>Btcrq</term><term>Btcrq iels</term><term>Btcry</term><term>Cd4q</term><term>Cd4y</term><term>Cd8a</term><term>Cd8aq</term><term>Cd8ay</term><term>Cd8q</term><term>Cell number</term><term>Cell populations</term><term>Cell receptor</term><term>Clone</term><term>Comparable results</term><term>Cytometric</term><term>Differential expression</term><term>Epithelial</term><term>Epithelial cells</term><term>Epithelium</term><term>Fangmann</term><term>Flow cytometric analysis</term><term>Flow cytometry</term><term>Fluorescence intensity</term><term>Gdtcrq</term><term>Gdtcrq cells</term><term>Gdtcrq iels</term><term>Helgeland</term><term>High density</term><term>Horizontal axis</term><term>Iels</term><term>Immunol</term><term>Immunological</term><term>Immunological methods</term><term>Individual animals</term><term>Individual samples</term><term>Insets show fluorescence profiles</term><term>Intestinal</term><term>Intestinal epithelial cells</term><term>Intestine</term><term>Intraepithelial</term><term>Intraepithelial lymphocytes</term><term>Isolation procedure</term><term>Isotype control immunoglobulin</term><term>Kearsey</term><term>Lamina</term><term>Lamina propria</term><term>Large numbers</term><term>Lefrancois</term><term>Light microscopy</term><term>Lymph</term><term>Lymph node cells</term><term>Lymphocyte</term><term>Lymphoid</term><term>Lymphoid cells</term><term>Major subsets</term><term>Methodology</term><term>Middle panel</term><term>National institutes</term><term>Node</term><term>Nylon</term><term>Nylon wool</term><term>Nylon wool purification</term><term>Patch lymphocytes</term><term>Percoll</term><term>Percoll solution</term><term>Peripheral lymph node cells</term><term>Phenotype</term><term>Phenotypic</term><term>Phenotypic characteristics</term><term>Previous reports</term><term>Propria</term><term>Receptor</term><term>Representative profiles</term><term>Right panel</term><term>Rt6q cells</term><term>Same cell populations</term><term>Small intestine</term><term>Stadnyk</term><term>Subset</term><term>Takimoto</term><term>Teitelbaum</term><term>Third fluorochrome</term><term>Todd</term><term>Total iels</term><term>Unique staining patterns</term><term>Vertical axis</term><term>Waite</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Intraepithelial lymphocytes (IELs) play critical roles in gut immunity. In mice, γδ T cells are a large component of the IEL population. In the rat, γδ 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 αβ T cell receptor (TcR). Among the αβTcR− cells was a population of γδ 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.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Massachusetts</li></region></list><tree><country name="États-Unis"><region name="Massachusetts"><name sortKey="Todd, Derrick" sort="Todd, Derrick" uniqKey="Todd D" first="Derrick" last="Todd">Derrick Todd</name></region><name sortKey="Bortell, Rita" sort="Bortell, Rita" uniqKey="Bortell R" first="Rita" last="Bortell">Rita Bortell</name><name sortKey="Greiner, Dale L" sort="Greiner, Dale L" uniqKey="Greiner D" first="Dale L" last="Greiner">Dale L. Greiner</name><name sortKey="Mordes, John P" sort="Mordes, John P" uniqKey="Mordes J" first="John P" last="Mordes">John P. Mordes</name><name sortKey="Rossini, Aldo A" sort="Rossini, Aldo A" uniqKey="Rossini A" first="Aldo A" last="Rossini">Aldo A. Rossini</name><name sortKey="Singh, Amrik J" sort="Singh, Amrik J" uniqKey="Singh A" first="Amrik J" last="Singh">Amrik J. Singh</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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