Abstract
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-gamma 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.
Publication types
-
Comparative Study
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Cell Differentiation / genetics
-
Cell Differentiation / immunology
-
Cells, Cultured
-
Cytokines / biosynthesis*
-
DNA-Binding Proteins / biosynthesis
-
DNA-Binding Proteins / genetics
-
GATA3 Transcription Factor
-
Humans
-
Interleukin-10 / biosynthesis
-
Interleukin-13 / biosynthesis
-
Interleukin-5 / biosynthesis
-
Interphase / genetics
-
Interphase / immunology
-
Lymphocyte Activation / genetics
-
Male
-
Protein Isoforms / biosynthesis
-
Protein Isoforms / genetics
-
Proto-Oncogene Proteins / biosynthesis
-
Proto-Oncogene Proteins / genetics
-
Proto-Oncogene Proteins c-maf
-
RNA, Messenger / biosynthesis
-
T-Box Domain Proteins
-
T-bet Transcription Factor
-
Th1 Cells / cytology*
-
Th1 Cells / immunology*
-
Th1 Cells / metabolism
-
Th2 Cells / cytology*
-
Th2 Cells / immunology*
-
Th2 Cells / metabolism
-
Trans-Activators / biosynthesis
-
Trans-Activators / genetics
-
Transcription Factors / biosynthesis
-
Transcription Factors / genetics
-
Up-Regulation / genetics
-
Up-Regulation / immunology
Substances
-
Cytokines
-
DNA-Binding Proteins
-
ETV5 protein, human
-
GATA3 Transcription Factor
-
GATA3 protein, human
-
Interleukin-13
-
Interleukin-5
-
MAF protein, human
-
Protein Isoforms
-
Proto-Oncogene Proteins
-
Proto-Oncogene Proteins c-maf
-
RNA, Messenger
-
T-Box Domain Proteins
-
T-bet Transcription Factor
-
Trans-Activators
-
Transcription Factors
-
Interleukin-10