Serine protease inhibition attenuates rIL-12-induced GZMA activity and proinflammatory events by modulating the Th2 profile from estrogen-treated mice.
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.
Department of Veterinary Biosciences and Diagnostic Imaging (E.K., T.L.G., S.D.H., R.M.G.), and Department of Small Animal Medicine and Surgery (C.W.S., M.H.), College of Veterinary Medicine, University of Georgia, Athens, Georgia 30602; and Immune Effector Cells Group (J.P.), Aragon Health Research Institute (IIS Aragon), Biomedical Research Centre of Aragon (CIBA), Nanoscience Institute of Aragon (INA), Aragon I+D Foundation (ARAID), University of Zaragoza, 50009 Zaragoza, Spain.
Ebru
E
Karpuzoglu
Chad W
CW
Schmiedt
Julian
J
Pardo
Megan
M
Hansen
Tai L
TL
Guo
Steven D
SD
Holladay
Robert M
RM
Gogal
D000818:Animals; D016207:Cytokines / Q000378:metabolism; D004967:Estrogens / Q000502:physiology*; D005260:Female; D050990:GATA3 Transcription Factor / Q000378:metabolism; D053804:Granzymes / Q000378:metabolism / Q000502:physiology*; D018836:Inflammation Mediators / Q000502:physiology*; D018664:Interleukin-12 / Q000502:physiology*; D015847:Interleukin-4 / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D051257:Proto-Oncogene Proteins c-maf / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000378:metabolism; D050800:STAT6 Transcription Factor / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013450:Sulfones / Q000494:pharmacology; D018418:Th2 Cells / Q000201:enzymology / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology
D016207:Cytokines; D004967:Estrogens; D050990:GATA3 Transcription Factor; C494748:Gata3 protein, mouse; D018836:Inflammation Mediators; C495608:Maf protein, mouse; D051257:Proto-Oncogene Proteins c-maf; D050799:STAT5 Transcription Factor; D050800:STAT6 Transcription Factor; D015842:Serine Proteinase Inhibitors; C494100:Stat6 protein, mouse; D013450:Sulfones; D018664:Interleukin-12; D015847:Interleukin-4; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D053804:Granzymes