FLOW CYTOMETRIC MEASUREMENT OF CALCIUM INFLUX IN MURINE T-CELL HYBRIDS USING FLUO-3 AND AN ORGANIC-ANION TRANSPORT INHIBITOR

被引：19

作者：

BAUS, E ^{[1
]}

URBAIN, J ^{[1
]}

LEO, O ^{[1
]}

ANDRIS, F ^{[1
]}

机构：

[1] UNIV LIBRE BRUXELLES, PHYSIOL ANIM LAB, B-1640 RHODE ST GENESE, BELGIUM

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1994年 / 173卷 / 01期

关键词：

CALCIUM INFLUX; FLOW CYTOMETRY; FLUO-3; T CELL HYBRID; SULFINPYRAZONE;

D O I：

10.1016/0022-1759(94)90281-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method is described to facilitate flow cytometric analysis of calcium mobilization upon stimulation of murine T cell hybrids. In these transformed cell lines, the accuracy of cytometric measurement of free cytoplasmic calcium with Fluo-3 is compromised by the rapid loss of the intracellular dye. We have found that the addition of sulfinpyrazone, a known organic-anion transporter inhibitor in epithelial cells and in macrophages, severely impairs the leakage of the Fluo-3 probe from the cytoplasmic matrix. Under appropriate conditions, sulfinpyrazone has little effect on the cell physiology and permits the detection of calcium influx in a variety of murine T cell hybrids.

引用

页码：41 / 47

页数：7

共 16 条

[1]

ANDRIS F, 1989, IMMUNOLOGY, V68, P1

[2] GENETIC AND MUTATIONAL ANALYSIS OF THE T-CELL ANTIGEN RECEPTOR [J].

ASHWELL, JD ;

KLAUSNER, RD .

ANNUAL REVIEW OF IMMUNOLOGY, 1990, 8 :139-167

[3] IDENTIFICATION OF A MURINE MONOCLONAL-ANTIBODY SPECIFIC FOR AN ALLOTYPIC DETERMINANT ON MOUSE CD3 [J].

COULIE, PG ;

UYTTENHOVE, C ;

WAUTERS, P ;

MANOLIOS, N ;

KLAUSNER, RD ;

SAMELSON, LE ;

VANSNICK, J .

EUROPEAN JOURNAL OF IMMUNOLOGY, 1991, 21 (07) :1703-1709

[4] FLOW CYTOMETRIC ANALYSIS OF TRANSPORT ACTIVITY IN LYMPHOCYTES ELECTROPORATED WITH A FLUORESCENT ORGANIC ANION DYE [J].

DINCHUK, JE ;

KELLEY, KA ;

CALLAHAN, GN .

JOURNAL OF IMMUNOLOGICAL METHODS, 1992, 155 (02) :257-265

[5]

DIVIRGILIO F, 1989, METHOD CELL BIOL, V31, P453

[6]

GRYNKIEWICZ G, 1985, J BIOL CHEM, V260, P3440

[7] THE MAJOR HISTOCOMPATIBILITY COMPLEX-RESTRICTED ANTIGEN RECEPTOR ON T-CELLS .1. ISOLATION WITH A MONOCLONAL-ANTIBODY [J].

HASKINS, K ;

KUBO, R ;

WHITE, J ;

PIGEON, M ;

KAPPLER, J ;

MARRACK, P .

JOURNAL OF EXPERIMENTAL MEDICINE, 1983, 157 (04) :1149-1169

[8] THE FINE SPECIFICITY OF ANTIGEN AND IA DETERMINANT RECOGNITION BY T-CELL HYBRIDOMA CLONES SPECIFIC FOR PIGEON CYTOCHROME-C [J].

HEDRICK, SM ;

MATIS, LA ;

HECHT, TT ;

SAMELSON, LE ;

LONGO, DL ;

HEBERKATZ, E ;

SCHWARTZ, RH .

CELL, 1982, 30 (01) :141-152

[9] LYMPHOCYTE-T ACTIVATION - THE ROLE OF PROTEIN-KINASE-C AND THE BIFURCATING INOSITOL PHOSPHOLIPID SIGNAL TRANSDUCTION PATHWAY [J].

ISAKOV, N ;

MALLY, MI ;

SCHOLZ, W ;

ALTMAN, A .

IMMUNOLOGICAL REVIEWS, 1987, 95 :89-111

[10]

ISEKI R, 1991, J IMMUNOL, V147, P4286

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