Document 0622
 DOCN  M9650622
 TI    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).
 DT    9605
 AU    Stanciu LA; Shute J; Holgate ST; Djukanovic R; Immunopharmacology Group,
       General Hospital, Southampton, UK.
 SO    J Immunol Methods. 1996 Jan 16;189(1):107-15. Unique Identifier :
       AIDSLINE MED/96163550
 AB    Highly enriched CD4+ and CD8+ human T cells were obtained from
       peripheral blood using a relatively simple and inexpensive method
       consisting of four steps: separation of mononuclear cells on Lymphoprep,
       removal of adherent monocytes by incubation in plastic petri dishes,
       removal of B cells, NK cells and further depletion of nonadherent
       monocytes by panning with anti-CD19, CD16, -CD14, -CD11b and -CD33 mAb,
       and separation of CD4+ and CD8+ T lymphocytes by magnetic cell sorting
       (MACS). Cell culture for up to 48 h showed preservation of function by
       both positively and negatively selected cells as determined by
       production of IL-8. Although the cell separation procedure had no effect
       on interleukin-2 receptor (IL-2R, CD25) expression, it induced
       production of IL-4 by both T cell subsets selected positively, implying
       cell activation by ligation of CD4 and CD8 molecules. Irrespective of
       the mode of separation, CD8+ T cells produced more IL-4, a cytokine
       which is associated with a Th2-type cytokine profile of CD4+ T cells. We
       conclude that our method for separating T cells into their CD4+ and CD8+
       subsets results in high cell purities with preservation of function, as
       determined by cytokine generation. If enriched cells are to be used for
       functional studies we recommend isolation by negative selection which
       has less effect on cell function. The relevance of the finding that CD8+
       T cells can be an important source of IL-4 remains to be elucidated.
 DE    Cell Adhesion/IMMUNOLOGY  Cell Survival/IMMUNOLOGY  Comparative Study
       Concanavalin A/PHARMACOLOGY  CD4-Positive T-Lymphocytes/*METABOLISM
       CD8-Positive T-Lymphocytes/*METABOLISM  Flow Cytometry  Human
       *Immunomagnetic Separation/METHODS  Interleukin-4/*BIOSYNTHESIS
       Interleukin-8/*BIOSYNTHESIS  Monocytes/IMMUNOLOGY
       Phytohemagglutinins/PHARMACOLOGY  JOURNAL ARTICLE

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       protected by Copyright Law (Title 17, U.S.Code).