As it has been established that demethylation of lysine 27 of histone H3 by the lysine-specific demethylase JMJD3 increases immune responses and thus elicits inflammation, we hypothesize that inhibition of JMJD3 may attenuate autoimmune disorders. We found that in vivo administration of GSK-J4, a selective inhibitor of JMJD3 and UTX, ameliorates the severity of experimental autoimmune encephalomyelitis (EAE). In vitro experiments revealed that the anti-inflammatory effect of GSK-J4 was exerted through an effect on dendritic cells (DCs), promoting a tolerogenic profile characterized by reduced expression of costimulatory molecules CD80/CD86, an increased expression of tolerogenic molecules CD103 and TGF-β1, and reduced secretion of proinflammatory cytokines IL-6, IFN-γ, and TNF. Adoptive transfer of GSK-J4-treated DCs into EAE mice reduced the clinical manifestation of the disease and decreased the extent of inflammatory CD4+ T cells infiltrating the central nervous system. Notably, Treg generation, stability, and suppressive activity were all exacerbated by GSK-J4-treated DCs without affecting Th1 and Th17 cell production. Our data show that GSK-J4-mediated modulation of inflammation is achieved by a direct effect on DCs and that systemic treatment with GSK-J4 or adoptive transfer of GSK-J4-treated DCs ex vivo may be promising approaches for the treatment of inflammatory and autoimmune disorders.

Following infection withThe female reproductive tract is an immunologically unique site which must respond to a diverse array of sexually transmitted pathogens and must also be tolerant to allogeneic sperm and to conceptuses. Pelvic inflammatory disease (PID) is an acute clinical syndrome associated with the ascending spread of microorganisms through the female reproductive tract (80). PID encompasses a multitude of inflammatory conditions of the upper reproductive tract organs, with the majority of proven cases of PID being caused by Chlamydia trachomatis and Neisseria gonorrhoeae (gonococcus) (32), and coinfection with both pathogens is common.Neisseria gonorrhoeae is the etiologic agent of gonorrhea, and the organism infects the mucosal epithelia of the male urethra and the lower genital tracts (vagina/cervix) of women. Localized infection with gonococci leads to a mucopurulent cervicitis in women, but it is also frequently asymptomatic. However, in approximately 10 to 25% (7,26,70) of untreated individuals, infection may ascend into the upper reproductive tract to involve the endometrium, ovaries, myometrium, parametrium, and Fallopian tubes (FT) (32, 46). The host response to this ascending infection is manifested as endometritis, pelvic (tubal or ovarian) peritonitis, tubal abscess, and salpingitis in the FT, and all of these inflammatory conditions encompass the clinical syndrome of PID. Long-term sequelae that develop in individuals presenting with PID, such as chronic pelvic pain, tubal damage, and ectopic pregnancy (7,26,70), are recognized as important public health problems worldwide (32,46).The FT is essentially a muscular organ whose lumen is lined by columnar ciliated cells and secretory cells with microvilli (68), and it plays a critical role in mammalian reproduction, functioning as a channel and storage organ for spermatozoa, a collecting vessel for oocytes released from the ovaries, the site of fertilization and zygote formation, and a means for transporting the early embryo to the uterus (54, 68). It is recognized that salpingitis induced by gonococcal infection causes significant tissue damage in the FT, which is resolved by a process of repair by infiltrating fibroblasts that leads to scarring. These events cause functional impairment of the tubes and irreversible infertility (80). However, little is known of the molecular mechanisms involved in the early stages of infection of the FT by ascending gonococci that initiate the inflammatory response. Studying the pathogenesis of gonococcus-induced salpingitis has relied on the use of ex vivo human FT organ tube

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Adoptive transfer of CD4+CD25+FOXP3+ regulatory T cells (Treg cells) has been successfully utilized to treat graft versus host disease and represents a promising strategy for the treatment of autoimmune diseases and transplant rejection. The aim of this study was to evaluate the effects of all-trans retinoic acid (atRA) and rapamycin (RAPA) on the number, phenotype, homing markers expression, DNA methylation, and function of induced human Treg cells in short-term cultures. Naive T cells were polyclonally stimulated and cultured for five days in the presence of different combinations of IL-2, TGF-β1, atRA and RAPA. The resulting cells were characterized by the expression of FOXP3, activation, surface and homing markers. Methylation of the Conserved Non-coding Sequence 2 was also evaluated. Functional comparison of the different culture conditions was performed by suppression assays in vitro. Culturing naive human T cells with IL-2/TGFβ1 resulted in the generation of 54.2% of Treg cells (CD4+CD25+FOXP3+) whereas the addition of 100 nM atRA increased the yield of Treg cells to 66% (p = 0.0088). The addition of RAPA did not increase the number of Treg cells in any of these settings. Treg cells generated in the presence of atRA had an increased expression of the β7 integrin to nearly 100% of the generated Treg cells, while RAPA treated cells showed enhanced expression of CXCR4. The differential expression of homing molecules highlights the possibility of inducing Treg cells with differential organ-specific homing properties. Neither atRA nor RAPA had an effect on the highly methylated CNS2 sites, supporting reports that their contribution to the lineage stability of Treg cells is not mediated by methylation changes in this locus. Treg cells generated in the presence of RAPA show the most potent suppression effect on the proliferation of effector cells.

Background: Infection of the Fallopian tubes (FT) by Neisseria gonorrhoeae (Ngo) can lead to acute salpingitis, an inflammatory condition resulting in damage primarily to the ciliated cells, with loss of ciliary activity and sloughing of the cells from the epithelium. Recently, we have shown that Ngo infection induced apoptosis in FT epithelium cells by a TNF-α dependent mechanism that could contribute to the cell and tissue damage observed in gonococcal salpingitis. Aim: To investigate the apoptosis-related genes expressed during apoptosis induction in cultured FT epithelial cells infected in vitro by Ngo. Materials and Methods: In the current study, we used cDNA macroarrays and real time PCR to identify and determine the expression levels of apoptosis related genes during the in vitro gonococci infection of FT epithelial cells. Results: Significant apoptosis was induced following infection with Ngo. Macroarray analysis identified the expression of multiple genes of the TNF receptor family (TNFRSF1B,-4,-6,-10A,-10B and-10D) and the Bcl-2 family (BAK1, BAX, BLK, HRK and MCL-1) without differences between controls and infected cells. This lack of difference was confirmed by RT-PCR of BAX, Bcl-2, TNFRS1A (TNFR-I) and TNFRSF1B (TNFR-II). Conclusion: Several genes related to apoptosis are expressed in primary cultures of epithelial cells of the human Fallopian tube. Infection with Ngo induces apoptosis without changes in the pattern of gene expression of several apoptosis-related genes. Results strongly suggest that Ngo regulates apoptosis in the FT by posttranscriptional mechanisms that need to be further addressed.

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