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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions |
umls-concept:C0016263,
umls-concept:C0024204,
umls-concept:C0025663,
umls-concept:C0025914,
umls-concept:C0026809,
umls-concept:C0060505,
umls-concept:C0079603,
umls-concept:C0080202,
umls-concept:C0178719,
umls-concept:C0184511,
umls-concept:C0205174,
umls-concept:C0596235,
umls-concept:C0936012,
umls-concept:C2348693
|
| pubmed:issue |
3
|
| pubmed:dateCreated |
1997-1-2
|
| pubmed:abstractText |
A visible-light, dual-laser, flow cytometric method was developed for the simultaneous analysis of intracellular ionized calcium concentration ([Ca2+]i) and three cell-surface markers (CD4, CD8, and Thy-1.2 antigens) by using the calcium probe fluo-3 and using R-phycoerythrin (PE), peridinin chlorophyll-alpha protein (PerCP), and allophycocyanin (APC) conjugated monoclonal antibodies (MoAbs). This improved method was used in the analysis of [Ca2+]i mobilization upon in vitro stimulation with mitogenic lectins [phytohaemagglutinin (PHA) or concanavalin A (ConA)], anti-CD3 MoAbs, or A23187 calcium ionophore in the heterogeneous lymph node cell populations from healthy C57BL/Ka mice. The present results show that the calcium responses were heterogeneous and dependent on the cellular immunophenotype, not only on lectins or anti-CD3 MoAbs stimulation, but also on the receptor-independent A23187 ionophore stimulation. An in situ fluo-3 calibration method (using A23187 and metabolic poisons in Ca2+ /EGTA buffers with known free calcium concentrations) indicated a resting [Ca2+]i in lymphocytes of 103 +/- 23 nM (mean +/- S.D.) but with significant differences between the [Ca2+]i in B cells and in all of the T-cell subsets (CD4+Thy-1+, CD4+Thy-1-, and CD8+T cells). Both the B cells and the T-cell subsets showed an increase of fluo-3 fluorescence upon in vitro stimulation with ConA or PHA, but the calcium mobilization following lectin stimulation was time delayed in all T-cell subsets. Only the T cells, including the CD4+Thy-1- subset, responded to anti-CD3 MoAbs. The percentage of responding cells upon stimulation with ConA was higher in T cells than in B cells. By contrast, PHA gave a higher response in B cells. After stimulation with different mitogens, [Ca2+]i increased in both CD4+ and CD8+ T-cell subsets. However, the percentage of responding cells was far higher in the CD4+Thy-1+ subset than in the CD4+Thy-1- or the CD8+T-cell subsets. The stimulation with A23187 ionophore induced a higher calcium response in B cells than in T cells. Interestingly, it also induced greater Ca2+ mobilization in CD4+ than in CD8+T cells. These results demonstrate the potential use of fluo-3 simultaneously with three fluorescein (FITC)-compatible fluorochromes. This technique may be useful for investigating the role of the CD4+Thy-1-T cells, a rare subset that is abnormally expanded in a murine acquired immunodeficiency syndrome (murine AIDS).
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Aniline Compounds,
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Thy-1,
http://linkedlifedata.com/resource/pubmed/chemical/Calcimycin,
http://linkedlifedata.com/resource/pubmed/chemical/Calcium,
http://linkedlifedata.com/resource/pubmed/chemical/Carotenoids,
http://linkedlifedata.com/resource/pubmed/chemical/Fluo-3,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescent Dyes,
http://linkedlifedata.com/resource/pubmed/chemical/Ionophores,
http://linkedlifedata.com/resource/pubmed/chemical/Lectins,
http://linkedlifedata.com/resource/pubmed/chemical/Mitogens,
http://linkedlifedata.com/resource/pubmed/chemical/Phycocyanin,
http://linkedlifedata.com/resource/pubmed/chemical/Phycoerythrin,
http://linkedlifedata.com/resource/pubmed/chemical/Protozoan Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Xanthenes,
http://linkedlifedata.com/resource/pubmed/chemical/allophycocyanin,
http://linkedlifedata.com/resource/pubmed/chemical/peridinin chlorophyll-a protein...
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
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| pubmed:issn |
0196-4763
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
1
|
| pubmed:volume |
23
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| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
205-17
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:8974866-Aniline Compounds,
pubmed-meshheading:8974866-Animals,
pubmed-meshheading:8974866-Antigens, Thy-1,
pubmed-meshheading:8974866-B-Lymphocytes,
pubmed-meshheading:8974866-CD4-Positive T-Lymphocytes,
pubmed-meshheading:8974866-CD8-Positive T-Lymphocytes,
pubmed-meshheading:8974866-Calcimycin,
pubmed-meshheading:8974866-Calcium,
pubmed-meshheading:8974866-Carotenoids,
pubmed-meshheading:8974866-Flow Cytometry,
pubmed-meshheading:8974866-Fluorescent Dyes,
pubmed-meshheading:8974866-Ionophores,
pubmed-meshheading:8974866-Lectins,
pubmed-meshheading:8974866-Lymph Nodes,
pubmed-meshheading:8974866-Male,
pubmed-meshheading:8974866-Mice,
pubmed-meshheading:8974866-Mice, Inbred C57BL,
pubmed-meshheading:8974866-Mitogens,
pubmed-meshheading:8974866-Phycocyanin,
pubmed-meshheading:8974866-Phycoerythrin,
pubmed-meshheading:8974866-Protozoan Proteins,
pubmed-meshheading:8974866-Stimulation, Chemical,
pubmed-meshheading:8974866-T-Lymphocytes,
pubmed-meshheading:8974866-Xanthenes
|
| pubmed:year |
1996
|
| pubmed:articleTitle |
Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets.
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| pubmed:affiliation |
Laboratory of Pathological and Cytological Anatomy, University Hospital of Liege, Belgium.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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