Article (Scientific journals)
Efficient Immunoselection of Cytolytic Effectors with a Magnetic Cell Sorter
Jacobs, Nathalie; MOUTSCHEN, Michel; Boniver, Jacques et al.
1993In Research in Immunology, 144 (2), p. 141-50
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Keywords :
Magnetic sorting; T-cell
Abstract :
[en] This paper describes a rapid and efficient method for the sorting of in vitro activated cytolytic effectors cells. For cytotoxic assays, a large number of cells with conserved function must be rapidly obtained. Immunomagnetic sorting was chosen because it is faster than flow cytometry sorting. The MACS system requires the use of paramagnetic beads of small diameter (100-150 nm), reputed to interfere minimally with cell function. In order to generate the cytolytic effectors, peripheral blood lymphocytes were cultivated in the presence of interleukin-2 (50 U/ml) and anti-CD3 monoclonal antibody (BMA030, 100 ng/ml) for 4 days. Cell separation was based on the membrane expression of the CD3 complex. The purity obtained for positive (CD3+) cell sorting with the MACS was higher than 95%. The purity of negative (CD3-) cell fraction was more variable, but further purification by flow cytometry rapidly yielded purity higher than 95%. Cytotoxic assays were performed against four target cell lines (K562, Daudi, HL60 and U937) and proliferation assays showed that both negatively and positively selected populations had conserved their function acquired during culture in the presence of anti-CD3 mAb and IL2.
Disciplines :
Immunology & infectious disease
Author, co-author :
Jacobs, Nathalie  ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
MOUTSCHEN, Michel  
Boniver, Jacques ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Greimers, Roland ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Schaaf-Lafontaine, Nicole ;  Université de Liège - ULiège > Department of Pathology
Language :
English
Title :
Efficient Immunoselection of Cytolytic Effectors with a Magnetic Cell Sorter
Publication date :
February 1993
Journal title :
Research in Immunology
ISSN :
0923-2494
Publisher :
Elsevier, Netherlands
Volume :
144
Issue :
2
Pages :
141-50
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
Oeuvre Belge du Cancer
CAC - Centre anticancéreux près l'Université de Liège asbl [BE]
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