SpringerProtocols: Abstract: 11. Isolation of T-Cell Subsets by Magnetic Cell Sorting (MACS)

11. Isolation of T-Cell Subsets by Magnetic Cell Sorting (MACS)

By: Luminita A. Stanciu², Ratko Djukanović²

Abstract

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T cells are defined by the expression of CD3 molecules on the cell surface in association with the T-cell receptor (TCR) to form the TCR/CD3 complex. Within the CD3⁺ cell population there are two main subsets identified by the expression of CD4 (CD4⁺ T cells) and CD8 (CD8⁺ T cells) cell surface molecules. In addition, there is a small percentage of CD3⁺ T cells which are either negative for both CD4 and CD8 (CD4⁻CD8⁻) or are double positive (CD4⁺CD8⁺).

Affiliation(s): (2) Southampton University General Hospital, Immunopharmacology Group, Southampton University, Southampton, England, UK

Book Title: T Cell Protocols: Development and Activation

Series: Methods in Molecular Biology | Volume: 134 | Pub. Date: Dec-22-1999 | Page Range: 133-141 | DOI: 10.1385/1-59259-682-7:133

Subject: Immunology

References

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1.	Stanciu, L. A., Shute, J., Promwong, C., Holgate, S. T., and Djukanović, R. (1996) Production of IL-8 and IL-4 by positively and negatively selected CD4⁺ and CD8⁺ human T cells following a four-step cell separation method including magnetic cell sorting (MACS). J. Immunol. Methods 189, 107–115.

2.	Stanciu, L. A., Shute, J., Promwong, C., Holgate, S. T., and, Djukanović, R. (1997) Increased levels of IL-4 in CD8⁺ T cells in atopic asthma. J. Allergy Clin. Immunol. 100, 373–378.

3.	Oyaizu, N., Chirmule, N., Kalyanaraman, V. S., Hall, W. W., Good, R. A., and Pahwa, S. (1990) Human immunodeficiency virus type 1 envelope glycoprotein gp120 produces immune defects in CD4⁺ T lymphocytes by inhibiting interleukin 2 mRNA. Proc. Natl. Acad. Sci. USA 87, 2379–2383.

4.	Goldman, F., Jensen, W. A. J., Johnson, G. L., Heasley, L., and Cambier, J. C. (1994) gp120 ligation of CD4 induces p56^lck activation and TCR desensitization independent of TCR tyrosine phosphorylation. J. Immunol. 153, 2905–2917.

5.	Ledbetter, J. A., June, C. H., Grosmaire, L. S., and Rabinovitch, P. S. (1987) Crosslinking of surface antigens causes mobilization of intracellular ionized calcium in T lymphocytes. Proc. Natl. Acad. Sci. USA 84, 1384–1388.

6.	Stumbles, P. and Mason, D. (1995) Activation of CD4⁺ T cells in the presence of a nondepleting monoclonal antibody to CD4 induces a Th2-type response in vitro. J. Exp. Med. 182, 5–13.

7.	Jabado, N., Le Deist, F., Fischer, A., and Hivroz, C. (1994) Interaction of HIV gp120 and anti-CD4 antibodies with the CD4 molecule on human CD4⁺ T cells inhibits the binding activity of NF-AT, NF-_κB and AP-1, three nuclear factors regulating interleukin-2 gene enhancer activity. Eur. J. Immunol. 24, 2646–2652.

8.	Miltenyi, S., Muller, W., Weichel, W., and Radbruch, A. (1990) High gradient magnetic cell separation with MACS. Cytometry 11, 231–238.

9.	Böyum, A., Lovhaug, D., Tresland, L., and Nordlie, E. M. (1991) Separation of leukocytes: improved cell purity by fine adjustments of gradient medium density and osmolarity. Scand. J. Immunol. 34, 697–712.

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