Serine protease inhibition attenuates rIL-12-induced GZMA activity and proinflammatory events by modulating the Th2 profile from estrogen-treated mice

Endocrinology. 2014 Aug;155(8):2909-23. doi: 10.1210/en.2014-1045. Epub 2014 May 19.


Estrogen has potent immunomodulatory effects on proinflammatory responses, which can be mediated by serine proteases. We now demonstrate that estrogen increased the extracellular expression and IL-12-induced activity of a critical member of serine protease family Granzyme A, which has been shown to possess a novel inflammatory persona. The inhibition of serine protease activity with inhibitor 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride significantly diminished enhanced production of proinflammatory interferon-γ, IL-1β, IL-1α, and Granzyme A activity even in the presence of a Th1-inducing cytokine, IL-12 from splenocytes from in vivo estrogen-treated mice. Inhibition of serine protease activity selectively promoted secretion of Th2-specific IL-4, nuclear phosphorylated STAT6A, signal transducer and activator of transcription (STAT)6A translocation, and STAT6A DNA binding in IL-12-stimulated splenocytes from estrogen-treated mice. Inhibition with 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride reversed the down-regulation of Th2 transcription factors, GATA3 and c-Maf in splenocytes from estrogen-exposed mice. Although serine protease inactivation enhanced the expression of Th2-polarizing factors, it did not reverse estrogen-modulated decrease of phosphorylated STAT5, a key factor in Th2 development. Collectively, data suggest that serine protease inactivity augments the skew toward a Th2-like profile while down-regulating IL-12-induced proinflammatory Th1 biomolecules upon in vivo estrogen exposure, which implies serine proteases as potential regulators of inflammation. Thus, these studies may provide a potential mechanism underlying the immunomodulatory effect of estrogen and insight into new therapeutic strategies for proinflammatory and female-predominant autoimmune diseases.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytokines / metabolism
  • Estrogens / physiology*
  • Female
  • GATA3 Transcription Factor / metabolism
  • Granzymes / metabolism
  • Granzymes / physiology*
  • Inflammation Mediators / physiology*
  • Interleukin-12 / physiology*
  • Interleukin-4 / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Proto-Oncogene Proteins c-maf / metabolism
  • STAT5 Transcription Factor / metabolism
  • STAT6 Transcription Factor / metabolism
  • Serine Proteinase Inhibitors / pharmacology*
  • Sulfones / pharmacology
  • Th2 Cells / enzymology
  • Th2 Cells / metabolism*
  • Up-Regulation / physiology


  • Cytokines
  • Estrogens
  • GATA3 Transcription Factor
  • Gata3 protein, mouse
  • Inflammation Mediators
  • Maf protein, mouse
  • Proto-Oncogene Proteins c-maf
  • STAT5 Transcription Factor
  • STAT6 Transcription Factor
  • Serine Proteinase Inhibitors
  • Stat6 protein, mouse
  • Sulfones
  • Interleukin-12
  • Interleukin-4
  • 4-(2-aminoethyl)benzenesulfonylfluoride
  • Granzymes