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Department of Respiratory Medicine and Allergy, GKT School of Medicine, King’s College, London, United Kingdom
We have developed an in vitro differentiation assay in which human
naive CD4+ cells are driven toward either the Th1 or Th2
phenotype. We have examined the interrelationships among the expression
of IL-2, IL-4, IL-5, IL-10, IL-13, GM-CSF, and IFN-
in individual
cells using intracellular cytokine staining at various times during the
differentiation process. We provide direct evidence that the Th2
cytokines IL-4, IL-5, and IL-13, unlike the other cytokines, are
regulated by a coordinated mechanism. We also show that IL-10 is
expressed by a different subset of cells that is prevalent at early
stages of Th2 differentiation, but then diminishes. Additionally we
demonstrate that while naive cells can express IL-2 upon activation,
they cannot express GM-CSF. Commitment to GM-CSF expression occurs
during differentiation in a Th1/Th2 subset-independent manner.
Furthermore, we have examined the levels of GATA3, c-Maf, T-bet, and
Ets-related molecule during human Th1/Th2 differentiation and
suggest that differences in the levels of these critical transcription
factors are responsible for commitment toward the Th1 or Th2
lineage.
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