An Infectious Disease–Associated Il12b Polymorphism Regulates IL-12/23 p40 Transcription Involving Poly(ADP-Ribose) Polymerase 1

Abstract

IL-12 and IL-23 are important host defense factors produced by APCs against certain intracellular and extracellular pathogens. Their dysregulation has also been implicated in several autoimmune diseases. The nucleotide polymorphism in the promoter region of (rs41292470 consisting of the long or short allele) encoding the shared subunit of IL-12 and IL-23, p40, has been reported to associate with susceptibility to infectious diseases and autoimmune disorders. How these genetic variants impact expression at the molecular level was unclear. We established an promoter-luciferase reporter system containing the long or short allele driving the reporter gene expression and found that the long allele (infection-resistant) displayed ∼2-fold higher transcriptional activity than the short allele (infection-susceptible), associated with a selective and differential nuclear binding activity to the two alleles in activated macrophages. DNA pull-down assays coupled with mass spectrometry analyses identified the specific DNA binding activity as poly(ADP-ribose) polymerase 1 (PARP-1). Small hairpin RNA-mediated knockdown of the endogenous PARP-1 expression resulted in reduced p40 mRNA expression and promoter activity. Bone marrow-derived macrophages from PARP-1-deficient mice had decreased p40 expression at both mRNA and protein levels. Furthermore, selective PARP-1 inhibitors resulted in impaired production of IL-12p40 and IL-23 in bone-marrow derived macrophages and PBMCs. Chromatin immunoprecipitation assay revealed that PARP-1 could bind specifically to in LPS-stimulated macrophages. Our study opens the way for further elucidating the molecular mechanism whereby allele-specific immune responses to foreign and self-antigens mediated by IL-12/IL-23 are controlled in an individually variable manner.

A novel ZRS variant causes preaxial polydactyly type I by increased sonic hedgehog expression in the developing limb bud

“…The primers are shown in Supplementary Table 4. A DNA pulldown assay was performed as described previously 22 using Caco-2 cells. For liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses, a Q-Exactive mass spectrometer (Thermo 588 Finnigan, San Jose, CA, USA) coupled with an Easy-nLC1000 HPLC system 589 (Thermo Fisher, San Jose, CA, USA) was used as described previously.…”

The Role of PARP1 in Monocyte and Macrophage Commitment and Specification: Future Perspectives and Limitations for the Treatment of Monocyte and Macrophage Relevant Diseases with PARP Inhibitors

“…Furthermore, the inhibition of ADP-ribosylation with PJ34 attenuated high glucose-induced macrophage calcification by inhibiting Runx2 expression and enhancing M2 differentiation. Moreover, IFN-γ and LPS/IFN-γ triggered the transcription of the IL12b gene encoding p40, a subunit of IL-12 and IL-23 in human peripheral blood mononuclear cells (PBMC), as well as in THP1-derived macrophages, in a PARP1-dependent fashion [85]. The enzyme recruitment to the IL12b locus resulted in promoter activation and p40 expression, whereas PARP1 silencing with shRNA resulted in the considerable reduction of p40 expression.…”

“…PARP1 regulates the transcription of IL10 in an allele-dependent way in macrophages engulfing apoptotic cells, but not upon LPS stimulation. It binds to -1082G>A alleles in the gene promoter, represses cytokine expression, and, thus, defines allele-dependent susceptibility to inflammatory pathology [85]. Furthermore, in cancer cells, PARP1 represses the anti-inflammatory cytokines IL10 and IL13 as well as PD-L1.…”

Enhancing Comprehensive Analysis of Secreted Glycoproteins from Cultured Cells without Serum Starvation

“…Among these, the deglycosylated peptide with the site of N303 from IL12B was up-regulated by 7.5 times, and this has been reported previously. 47,48 We also found that the two deglycosylated peptides with the site of N183 or N267 from intercellular adhesion molecule 1 (ICAM1) were up-regulated by 2.1 and 2.5 times, respectively. ICAM1 is a known cellsurface glycoprotein involved in leukocyte adhesion and is often up-regulated under inflammatory conditions.…”