Chemical agents and enzymes used for the extraction of gut lymphocytes influence flow cytometric detection of T cell surface markers

IF 1.7 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of immunological methods Pub Date : 2000-03-06 Epub Date: 2000-02-28 DOI:10.1016/S0022-1759(99)00243-4

Nancy Van Damme , Dominique Baeten , Martine De Vos , Pieter Demetter , Dirk Elewaut , Herman Mielants , Gust Verbruggen , Claude Cuvelier , Eric M. Veys , Filip De Keyser

{"title":"Chemical agents and enzymes used for the extraction of gut lymphocytes influence flow cytometric detection of T cell surface markers","authors":"Nancy Van Damme , Dominique Baeten , Martine De Vos , Pieter Demetter , Dirk Elewaut , Herman Mielants , Gust Verbruggen , Claude Cuvelier , Eric M. Veys , Filip De Keyser","doi":"10.1016/S0022-1759(99)00243-4","DOIUrl":null,"url":null,"abstract":"<div>Chemical agents (DTT, EDTA) and enzymes (collagenase, DNAse) are often used for the generation of a single cell suspension from tissue samples. In this study, flow cytometry was used to examine the effect of chemical agents and enzymes on the expression of cell membrane markers of T lymphocytes from tonsils and peripheral blood. Expression of CD4, CD8, CD25, CD38, L-selectin, CD44, αEβ7 and α4β7 were studied. Incubation of lymphocytes with DTT and EDTA resulted in a decrease of CD38, αEβ7 and α4β7 expression. Incubation with collagenase A and DNAse resulted in a decrease of CD25, L-selectin, αEβ7 and α4β7. The results of this study indicate that a careful interpretation is necessary for phenotypic descriptions of lymphocyte populations obtained by enzymatic isolation techniques.</div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"236 1","pages":"Pages 27-35"},"PeriodicalIF":1.7000,"publicationDate":"2000-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0022-1759(99)00243-4","citationCount":"38","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of immunological methods","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0022175999002434","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2000/2/28 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 38

Abstract

Chemical agents (DTT, EDTA) and enzymes (collagenase, DNAse) are often used for the generation of a single cell suspension from tissue samples. In this study, flow cytometry was used to examine the effect of chemical agents and enzymes on the expression of cell membrane markers of T lymphocytes from tonsils and peripheral blood. Expression of CD4, CD8, CD25, CD38, L-selectin, CD44, αEβ7 and α4β7 were studied. Incubation of lymphocytes with DTT and EDTA resulted in a decrease of CD38, αEβ7 and α4β7 expression. Incubation with collagenase A and DNAse resulted in a decrease of CD25, L-selectin, αEβ7 and α4β7. The results of this study indicate that a careful interpretation is necessary for phenotypic descriptions of lymphocyte populations obtained by enzymatic isolation techniques.

查看原文本刊更多论文

用于提取肠道淋巴细胞的化学试剂和酶影响T细胞表面标记物的流式细胞术检测

化学试剂(DTT, EDTA)和酶(胶原酶，DNAse)通常用于从组织样品中产生单细胞悬浮液。本研究采用流式细胞术检测化学试剂和酶对扁桃体和外周血T淋巴细胞细胞膜标志物表达的影响。观察CD4、CD8、CD25、CD38、l -选择素、CD44、αEβ7、α4β7的表达。DTT和EDTA孵育淋巴细胞导致CD38、αEβ7和α4β7表达降低。与胶原酶A和DNAse孵育导致CD25、l -选择素、αEβ7和α4β7的表达降低。本研究的结果表明，仔细的解释是必要的表型描述，通过酶分离技术获得淋巴细胞群。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of immunological methods 医学-免疫学

CiteScore

4.10

自引率

0.00%

发文量

120

审稿时长

3 months

期刊介绍： The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells. In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.