Distinctive calcineurin-dependent (FK506-sensitive) mechanisms regulate the production of the CC chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES vs IL-2 and TNF-alpha by activated human T cells.
Calcineurin (CaN) controls the production of multiple cytokines, including IL-2 and TNF-alpha, during T cell activation. However, its role in chemokine production is unclear. Here, we used the CaN inhibitor FK506 to probe for the contribution of CaN in MIP-1alpha, MIP-1beta, and RANTES production at the protein and mRNA levels in human T cells stimulated via CD3/PMA or CD3/CD28. With both modes of activation, FK506 inhibited RANTES production only partially and late during a 3-day culture, whereas it suppressed both MIP-1alpha and MIP-1beta production throughout the culture. However, FK506 inhibition was more pronounced on MIP-1beta than MIP-1alpha, especially in CD3/CD28-activated T cells. Surprisingly, FK506 also significantly reduced MIP-1beta induction by PMA alone. Furthermore, comparison with IL-2 and TNF-alpha revealed that both were more potently inhibited by the drug upon CD3/PMA or CD3/CD28 induction than either MIP-1alpha or MIP-1beta. These differences in FK506 sensitivity were also observed in CD4(+) and CD8(+) T cell subsets. Therefore, all three chemokines are affected by FK506 distinctly from one another and from IL-2 and TNF-alpha, suggesting that CaN participates to different extents in the induction of these cytokines during T cell activation. Further evidence that this induction relies on distinctive mechanisms, depending on the cytokine, came from analyses of the kinetics and cycloheximide sensitivity of cytokine mRNA expression.
Staruch
M J
MJ
Department of Immunology, Merck Research Laboratories, Rahway, New Jersey, 07065, USA.
Camacho
R
R
Dumont
F J
FJ
D018106:CD28 Antigens / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D019703:Calcineurin / Q000502:physiology*; D065095:Calcineurin Inhibitors; D002478:Cells, Cultured; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D018946:Chemokine CCL5 / Q000096:biosynthesis / Q000235:genetics; D003513:Cycloheximide / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D007376:Interleukin-2 / Q000096:biosynthesis; D008213:Lymphocyte Activation; D019402:Macrophage Inflammatory Proteins / Q000096:biosynthesis / Q000235:genetics; D008934:Mitogens / Q000378:metabolism / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism / Q000494:pharmacology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D018106:CD28 Antigens; D017252:CD3 Complex; D065095:Calcineurin Inhibitors; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D018946:Chemokine CCL5; D016207:Cytokines; D007376:Interleukin-2; D019402:Macrophage Inflammatory Proteins; D008934:Mitogens; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D003513:Cycloheximide; D019703:Calcineurin; D013755:Tetradecanoylphorbol Acetate; D016559:Tacrolimus