Analysis and sorting of T cells according to cytokine expression.
Functional distinct populations of T helper cells can be defined according to the expression of cytokines. A remarkable diversity of cytokine expression has been demonstrated in single T cells even from clonal populations and up to now no stable surface markers have been described which on the single-cell level directly correlate with the secretion of a certain cytokine. Since cytokines are the major parameters of T cell effector function we have developed strategies which now allow to separate cells according to the specific cytokines they secrete. The "affinity matrix technology"--secreted molecules are relocated to the cell surface by an artificially created antibody matrix--allows to isolate cells according to a distinct secreted product. In addition to this universal, but laborious technology, we could demonstrate by high-sensitivity immunofluorescence that IFN-gamma and IL-10 but not IL-2 and IL-4 are specifically expressed in low copy number on the surface of cells secreting these cytokines. Surface IFN-gamma and IL-10 are the first unambiguous surface markers for pro-inflammatory IFN-gamma-secreting Th 0/1 cells and IL-10-producing anti-inflammatory Th2/3 cells. We have used purified cytokine-secreting T cells to study in individual T cells the sequential production of IL-2, IFN-gamma, and IL-10, the stability of IFN-gamma expression and the selective homing of IFN-gamma-producing cells into inflamed tissues.
Scheffold
A
A
Deutsches Rheumaforschungszentrum Berlin, Germany.
Lohning
M
M
Richter
A
A
Assenmacher
M
M
Manz
R
R
Austrup
F
F
Hamann
A
A
Radbruch
A
A
D000818:Animals; D002469:Cell Separation / Q000379:methods; D016207:Cytokines / Q000096:biosynthesis* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D007371:Interferon-gamma / Q000096:biosynthesis / Q000378:metabolism; D016753:Interleukin-10 / Q000096:biosynthesis / Q000378:metabolism; D007700:Kinetics; D008565:Membrane Proteins / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D016176:T-Lymphocyte Subsets / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*
D016207:Cytokines; D008565:Membrane Proteins; D016753:Interleukin-10; D007371:Interferon-gamma