Positive selection of NK1.1+ cells on a magnetic cell separator (MACS).
Natural killer (NK) cells are involved not only in resistance to tumors and infection, but also in some transplantation reactions. A specific, inexpensive method for purifying large numbers of NK cells is often required. All NK cells in H-2b mice express the surface marker NK1.1. We report a method for positively selecting NK1.1+ spleen cells from normal and Poly I:C-stimulated C56Bl/6 mice using a magnetic cell separation technique known as MACS. Our results show that cytotoxic activity directed at YAC-1 target cells by normal and Poly I:C-stimulated spleen cells could be increased five-fold using this method. We also found that spleen cells from mice given Poly I:C could lyse NK-resistant, BW1100 target cells, and that this activity could be increased two-fold. Flow cytometry analysis of Poly I:C-stimulated, MACS enriched, NK1.1+ spleen cells revealed the presence of two subpopulations: one consisting of LGL and the other consisting of smaller, agranular lymphocytes (SAL). After enrichment, the percentage of NK1.1+ spleen cells increased from 69% to 91% in the LGL subpopulation and from 33% to 73% in the SAL subpopulation. These results clearly demonstrate the effectiveness of the MACS technique for purifying large numbers of NK1.1+ cells for both flow cytometric and functional analyses.
Ellison
C A
CA
University of Manitoba, Department of Immunology, Winnipeg, Canada.
Gartner
J G
JG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D005260:Female; D005434:Flow Cytometry; D006825:Hybridomas; D018189:Immunomagnetic Separation; D007694:Killer Cells, Natural; D016131:Lymphocyte Subsets; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal