These data corroborated with findings in a murine model, showing that protective efficacy following ID immunization requires a higher sporozoite inoculation compared to IV immunization, and suggesting

that differences in protective efficacy may be related to the number of sporozoites selleck reaching the liver. Using bioluminescent parasites, we studied the relation between parasite liver load following IV or ID sporozoite infection and protective immunity following IV or ID immunizations by P. berghei RAS or CPS protocols. Female BALB/cByJ and C57BL/6J, 8 weeks of age, were purchased from Elevage Janvier (Le Genest Saint Isle, France). All studies were performed according to the regulations of the Dutch “Animal On Experimentation act” and the European guidelines 86/609/EEG. Approval was obtained from the Radboud University Experimental Animal Ethical Committee (RUDEC 2009-019, RUDEC 2009-225). P. berghei (ANKA) sporozoites (spz) were obtained by dissection of

the salivary glands of infected female Anopheles stephensi mosquitoes 21–28 days after infected blood BMN 673 molecular weight meal. For radiation-attenuated sporozoites (RAS), infected mosquitoes were irradiated at 16 000 rad (Gammacell 1000 137Cs, Atomic Energy of Canada Ltd, Ontario, Canada) prior to dissection. All immunizations were performed with freshly isolated sporozoites. BALB/cByJ mice were immunized once with 50 000 P. berghei sporozoites. C57BL/6J mice Tobramycin received three injections of 10 000 sporozoites with 7 days intervals. Different immunization protocols were used given that in contrast to BALB/c mice, multiple sporozoite inoculations are required to induce protection in C57BL/6 mice (19,20). In both mouse strains, the choice for specific immunization dose was made based on a suspected (sub)optimal level of conferred protection. All immunizations were performed by IV injection (200 μL in the tail vein) or ID injection (50 μL in the proximal part of each hind leg). For the chloroquine prophylactic sporozoites (CPS) immunizations, mice received a daily i.p. injection

of 800 μg of chloroquine base starting simultaneously with the first sporozoite inoculation up to 2 weeks after the last sporozoite inoculation. Chloroquine diphosphate (CQ; Sigma-aldrich, Zwijndrecht, Netherlands) was diluted in PBS and administered to mice. At the end of the chloroquine treatment and 1 day before challenge, absence of parasitemia was confirmed by the examination of Giemsa-stained slides of tail blood. Groups of BALB/cByJ and C57BL/6J mice, immunized with either irradiated sporozoites or sporozoites under chloroquine cover, were challenged by Plasmodium berghei expressing firefly Luciferase and the green fluorescent protein (PbGFP-Luccon) infectious mosquito bites (5–11 mosquitoes) 2 weeks after the chloroquine treatment. Salivary glands of all blood-engorged mosquitoes were dissected to confirm the presence of sporozoites.

Characteristic PML lesions have been described as large, subcortical, grey-matter-sparing lesions appearing hyperintense on T2 and fluid-attenuated inversion recovery and hypointense

on T1 scans; contrast enhancement may occur [47]. The anti-CD52 mAb alemtuzumab (Lemtrada®) has been shown to be highly effective and is approved for active relapsing MS in Europe [10-12, 69]. Disease activity is defined as clinical or radiological deterioration [70]. Mechanisms of action include depletion of CD52-expressing T/B lymphocytes, natural killer (NK) cells, dendritic cells and monocytes/macrophages with skewed repopulation leading to a reprogramming of the immune repertoire [71, 72]. Already in earlier studies, patients especially with an early relapsing disease course appeared

to benefit most from alemtuzumab Fulvestrant chemical structure treatment, leading to the concept of a therapeutic window relatively early during the disease, when highly active immunotherapy may exert most profound effects [72]. This was reflected in the inclusion criteria for the pivotal Phase III trials Wnt antagonist CARE-MS I and II (Comparison of Alemtuzumab and Rebif® Efficacy in Multiple Sclerosis, Studies One and Two). CARE-MS I included active relapsing, therapy-naive MS patients, whereas CARE-MS II focused on relapsing MS refractory to first-line therapy [10, 12]. Especially in terms of disease progression, the latter patient group appeared to benefit most. Whereas current EMA approval is relatively broad [70], careful patient selection

is mandatory, as SADRs have been reported and thorough adherence to safety assessments is necessary. This is stressed by long-term data from the Phase II trial CAMMS223, with one additional SADR (Goodpasture syndrome), but also sustained reduction of disability accumulation and relapse rates compared to active comparator [73], revealing the dilemma of long-lasting efficacy versus potential SADRs. Alemtuzumab is applied intravenously with a first treatment cycle of 12 mg over 5 days, followed by a second therapy cycle over 3 days after 12 months [10, 12, 69]. Further cycles are not intended, but the question of when and how to continue DMD treatment after two cycles is unanswered. There is no class I evidence for different treatment protocols in this indication. During and for 1 month after treatment, acyclovir (200 mg twice daily) has to be administered prophylactically. C-X-C chemokine receptor type 7 (CXCR-7) Therapy surveillance with large treatment intervals, but necessarily close safety monitoring, will be a challenge in clinical practice [74] and emphasizes even more the importance of patient education, counselling and informed consent to assure adherence to safety measures. These include differential blood count, serum creatinine and urine analysis before first administration and monthly afterwards; regular testing of thyroid stimulating hormone (TSH) levels has to be performed before treatment initiation and every 3 months up to 4 years after the last administration [70].

3%) (Table 2). The results for MgEDTA–IPM and MgEDTA–CAZ were discordant for 16 MBL producers (Table 3). There were no false positive results for MgEDTA–IPM and MgEDTA–CAZ. Two P. aeruginosa carrying VIM-2 and one E. cloacae carrying IMP-1 had negative results with MgEDTA–IPM and MgEDTA–CAZ (Table 4); they were

also negative by the SMA disk method. However, two false negative P. aeruginosa became positive when biapenem and doripenem were used with Mg-EDTA, and one false negative E. cloacae became positive when panipenem and meropenem were used as substrates. After NDM-1 Dok01 was reported, two NDM-1-producing K. pneumoniae were identified by government-instigated phosphatase inhibitor library surveillance in Japan. These isolates were collected from elderly people who had not recently traveled abroad and had had no contact with the Indian subcontinent. Although NDM-1 producers from clinical isolates are rare in Japan, accurate screening methods to detect them are needed to prevent their further transmission in both hospitals and communities. Many clinical laboratories perform confirmatory tests for MBL production against carbapenem-resistant strains [20]. The DDST using SMA is the most convenient of the phenotypic MBL detection methods. However, the growth-inhibitory zone between IPM and the SMA disks is not large enough to be classified as positive with NDM-1 Dok01 [11]. In contrast to SMA disks, DDSTs using IPM disks and Mg-EDTA, Ca-EDTA,

Co-EDTA or Cu-EDTA detected two NDM-1 producers. In addition, the DDSTs using Mg-EDTA had high sensitivity (96.0%) and specificity (100%) for 75 MBL producers and 25 non-MBL producers. Galani et al. Selleckchem Roxadustat Methisazone reported that combined disk test with CAZ and EDTA (750 µg), and DDSTs with IPM disks 10 mm away from EDTA disks have high sensitivity (97.9–100%) and specificity (91.9–96%) in Enterobacteriaceae [14]. That we obtained similar sensitivity and specificity demonstrates that Mg-EDTA can be used as a MBL inhibitor.

Several reports have indicated that AmpC β-lactamase may cause false negative results in DDSTs using SMA [20, 21]. Arakawa et al. also reported that some MBL-producing gram-negative bacilli are difficult to detect. Because they have a low level of resistance to IPM, the expansion of the zone of inhibition is inconclusive [13]. In our study, only 3 of 75 strains were false negative by both MgEDTA–CAZ and MgEDTA–IPM; these three strains were also false negative in DDSTs using SMA. Two false negative P. aeruginosa strains were resistant to six carbapenems and one false negative E. cloacae was resistant to CAZ but susceptible to six carbapenems. Carbapenem resistance in P. aeruginosa is considered to be associated with loss of OprD outer membrane proteins and/or overexpression of active efflux systems in combination with strong expression of AmpC β-lactamase [22]. Furthermore, IPM induces expression of AmpC β-lactamase in P. aeruginosa more strongly than does doripenem [23].

IL-6, along with IL-8, has been shown to promote Treg migration to certain tumours [45] and may therefore play a crucial role in Treg signalling. A recent study in type II diabetes has also shown that the percentage of CD4(+)CD25(hi)CD127(–) Tregs are correlated positively SP600125 research buy with plasma IL-6 [46]. Mesenchymal stem cells from mice were shown to promote the differentiation

of uncommitted naive T cells to FoxP3+ regulatory T cells [47]. This study has shown that the major cytokines involved in these processes were transforming growth factor (TGF)-β and IL-6 and that immunomodulation could be blocked by administration of anti-TGF-β or anti-IL-6, suggesting paracrine mechanisms. In another study in mice, excessive Selleckchem Fludarabine IL-6 production caused an increase in natural Tregs, which maintained their immunosuppressive capacities both in vitro and in vivo [48]. This suggests that IL-6 may be a key mediator in effector/regulatory T cell balance. However, there are also data that are in contrast to our findings. In another study, using Tregs derived from the rheumatic synovium, blocking IL-6

or TNF increased the suppressive immunomodulatory capacities of these cells [49]. This study suggests that Treg function varies importantly with the inflammatory milieu in the joint. These findings are not in accordance with our experiments, in which IL-6 maintained the percentage of Tregs. This discrepancy may be due to differences in the underlying pathologies, but also reflect the dual role of IL-6. However, the functional consequences remain to be identified in upcoming experiments. We have chosen to carry out our study on Tregs taken from healthy donors in order to provide information TGF-beta inhibitor on the immunomodulatory

capacities of MSCs and ruling out possible influences of OA on Tregs and thus on Treg–MSC interaction. An important question is whether or not Tregs taken from OA patients differ in their interaction with MSCs. Our findings therefore raise several questions that need to be verified by future research. Understanding the effects of MSCs in local joint inflammation in OA may pave the way for future cell-based approaches, as MSCs can be supposed to not only exert their regenerative potential when administered, but also intervene in local immunity. This study has several limitations. First, a comparison to MSCs taken from healthy donors would be useful to detect whether the inability to recruit Tregs from a CD4+ population, as has been shown by other groups, should be interpreted as a reduced capacity of immunomodulation in OA. This question has not yet been addressed due to ethical questions, as synovium from healthy individuals is not easily available. Taking synovium from a young population undergoing hip or knee arthroscopy might falsify the results, as it is known that cartilage injuries are found in up to 65% of the patients during arthroscopy, irrespective of the underlying pathology [50].

These data showing a higher transcription

of il-17a, rorγt and il-22 genes in iNKT cells from NOD mice strengthen the differences in iNKT cells between this autoimmune strain and C57BL/6 mice. To determine whether iNKT17 cells infiltrate the pancreas of NOD mice, we have analyzed pancreatic infiltrates from NOD and Vα14 NOD transgenic mice that express iNKT cell characteristic TCRα chain and exhibit a10 fold increased frequency and number of iNKT cells in lymphoid tissues 6 as well as in the pancreas 29. iNKT17 cells represent 6% of all iNKT cells infiltrating the pancreas in NOD and Vα14 NOD mice (Fig. 2A). We next assessed whether this frequency varies at different stages of insulitis. At 6 wk of age NOD mice have a small infiltrate of hematopoietic cells, at 12-wk peri-insulitis is more abundant click here and at 20 wk many pancreatic islets are characterized by a destructive insulitis leading to diabetes onset 30. Indeed,

we observed an increased frequency of pancreatic infiltrating hematopoietic (CD45+) cells with aging (Fig. 2B). Even though, iNKT17 cell frequency among iNKT cells as well as iNKT cell frequency among CD45+ cells infiltrating pancreas remained stable (Fig. 2B), the number of iNKT17 cells increased with the enhanced infiltration of pancreas, meaning that they could participate in the destruction of islet cells. CCR6 and CD103 integrin expression has been described on iNKT17 cells 28 and CCR6 has been involved find more in the recruitment of pathogenic Th17 cells in CIA 23. All iNKT17 cells from ILNs are CD103+ and the level of CD103 expression is higher in iNKT17 cells of NOD mice as compared with C57BL/6 mice (Supporting Information Fig. 1). iNKT17 cells from ILNs are mainly CCR6+, whereas in PLNs and spleen only a fraction of iNKT17 cells express CCR6 and CD103 (Supporting Information Fig. 1). The analysis of CCR6 and CD103 expression these on pancreatic iNKT17 cells showed that, while 60% of iNKT17 cells expressed CD103 integrin, most of them were negative for CCR6 (Fig. 3C). These

data suggest that iNKT17 cell recruitment in the pancreas is independent of CCR6, whereas CD103 could play a role in the retention of these cells. To determine whether iNKT17 cells express IL-17A mRNA in the absence of exogenous stimulation such as PMA and ionomycin, iNKT cells were purified from the pancreas, PLNs and ILNs from Vα14 NOD mice. Expression of other genes usually associated with iNKT17 cells were also assessed by quantitative-PCR (Fig. 3D). IL-21 and IL-22 mRNA were barely detectable in the three organs analyzed. Interestingly, il-17a gene was expressed at much higher level in pancreatic iNKT cells than in iNKT cells from PLNs and ILNs (6- and 13-fold increased respectively). A similar trend was observed for il-17f gene. In contrast, rorγt and il-23r gene expression was not significantly different in iNKT cells from pancreas and ILNs.

The synergistic effect with nystatin was determined similarly. The effect of licorice compounds on biofilm formation was evaluated using a microplate assay and crystal violet staining. The effect of licorice compounds CYC202 on yeast-hyphal transition was determined by microscopic observation. The toxicity of licorice compounds towards oral epithelial cells was evaluated with an MTT assay. Glabridin and licochalcone A showed antifungal activity on C. albicans while glycyrrhizic acid had no effect. Complete growth inhibition occurred with sub-inhibitory concentrations

of nystatin with either glabridin or licochalcone A. Biofilm formation was inhibited by 35–60% in the presence of licochalcone A (0.2 μg ml−1). A strong inhibitory effect (>80%) on hyphal formation was observed with licochalcone A or glabridin (100 μg ml−1). Glabridin and licochalcone A at high concentrations showed toxicity towards oral epithelial cells. In summary, glabridin AMPK inhibitor and licochalcone

A are potent antifungal agents and may act in synergy with nystatin to inhibit growth of C. albicans. Licochalcone A has a significant effect on biofilm formation, while both licochalcone A and glabridin prevented yeast-hyphal transition in C. albicans. These results suggest a therapeutic potential of licochalcone A and glabridin for C. albicans oral infections. “
“Caspofungin is a member of the echinocandin class of antifungal compounds that inhibit 1,3-β-d-Glucan synthase. As patient exposure to caspofungin (CAS) broadens, the number of infecting strains with reduced susceptibility to this drug is expected to rise. In the present study, the in vitro effects of varying concentrations of CAS

against Candida albicans isolates presenting reduced susceptibility to CAS were studied in comparison with a reference strain. Two C. albicans isolates presenting high minimal inhibitory concentrations (MIC = 8 μg ml−1) were selected: one isolate obtained in the laboratory under continuous antifungal selection pressure (CaIn-R) and one clinical isolate (CaClin-R) from a patient with a therapeutic failure. Results showed that after 24 h of CAS exposure, CaIn-R and CaClin-R presented a partial growth inhibition in comparison with the reference strain. Moreover, scanning electron G protein-coupled receptor kinase microscopy and transmission electron microscopy studies showed that the cell walls of CaIn-R and CaClin-R were less altered than that of the reference strain. These observations suggested that although CaIn-R and CaClin-R cells were misshapen after CAS exposure, cell lysis was limited after 24 h of treatment indicating higher survival ability for CaIn-R and CaClin-R in the presence of CAS. “
“This study describes the isolation of Cryptococcus neoformans and Cryptococcus gattii from patients with chronic meningitis who were admitted to 16 Malaysian hospitals, from 2003 to 2004. Of the 96 cryptococcal cases reported over the 2-year period, 74 (77.1%) patients were male and 45 (46.

Patients with clinical suspicion of Akt molecular weight antifungal treatment failure need prompt workup for adequacy of treatment, focal sources of sustained infection and potential superinfection. “
“Accurate identification of fungal pathogens using a sequence-based approach requires an extraction method that yields template DNA pure enough for polymerase chain reaction (PCR) or other types of amplification. Therefore, the objective of this study was to develop and standardise a rapid,

inexpensive DNA extraction protocol applicable to the major fungal phyla, which would yield sufficient template DNA pure enough for PCR and sequencing. A total of 519 clinical and culture collection strains, comprised of both yeast and filamentous fungi, were prepared using our extraction method to determine its applicability for PCR, which targeted the ITS and D1/D2 regions in a single PCR amplicon. All templates were successfully amplified and found

to yield the correct strain identification when sequenced. This protocol could be completed in approximately 30 min and utilised a combination of physical and chemical extraction methods but did not require organic solvents nor ethanol precipitation. The method reduces the number of tube manipulations and yielded suitable template DNA for PCR amplification from all phyla that were tested. “
“Data on diagnostic performance of Galactomannan (GM) testing in patients under mould-active regimens are limited. Whether sensitivity of GM testing for diagnosing breakthrough invasive aspergillosis XL184 research buy (IA) is decreased under antifungal prophylaxis/therapy remains therefore a point of discussion. We retrospectively analysed GM test results in patients who were admitted with underlying 17-DMAG (Alvespimycin) HCl haematological malignancies to two Divisions of the Medical University Hospital of Graz, Austria, between 2009 and 2012. Only cases of probable and proven IA that were diagnosed by other methods than GM testing were included (time of diagnosis = day 0). We compared GM results of patients with/without therapy/prophylaxis for the period of 2 weeks prior (week −2) until

3 weeks postdiagnosis. A total of 76 GM test results in nine patients were identified. Six patients had received antifungal therapy/prophylaxis from week −2, whereas three patients were treated with therapy from the time of diagnosis at week 0. GM testing was positive in 45/76 (59%) of samples. Sensitivity of GM testing for detection of proven or probable IA at week −1 and 0 was 77% and 79% in patients with mould-active regimens. We conclude that GM testing might be a useful diagnostic method for breakthrough IA in patients receiving mould-active prophylaxis/therapy. “
“Poor susceptibility of Cryptococcus neoformans to fluconazole (FLC) is a matter of concern among clinicians in Africa. The emergence of resistance to FLC was recently reported in Kenya, but it is not known whether it is widespread.

Graft and patient survival were analysed using the Kaplan–Meier method. Pretransplant CMV D/R serostatus was available for the whole cohort, with EBV D/R serostatus available for 2566 transplants (56.8%).

Serostatus for both viruses was significantly associated with donor and recipient age and recipient smoking status. For both viruses the majority of transplants were in a D+/R+ serostatus setting: 45.3% for CMV and 77.9% for EBV. D/R serostatus for either virus did not have a significant effect on graft or patient survival. We conclude that in the current era of viral prophylaxis and surveillance, long-term outcome for the kidney transplant population is unaffected by D/R CMV and EBV serostatus. “
“A higher prevalence of sleep

apnoea (SA) has been observed in the chronic kidney disease (CKD) population compared with estimates in the general population. Increased rates of SA have been NVP-BEZ235 clinical trial described in patients with XL765 various renal-related diagnoses including dialysis, renal transplant, early-stage CKD and proteinuria. The mechanism or underlying aetiology for this association is different for each type of kidney disease. The extracellular fluid volume and metabolic derangements that characterize the uremic state likely contributes to SA in the dialysis population. SA causing direct renal insults from haemodynamic changes, ischaemic stress, or an intermediary condition such as hypertension, can lead to early CKD and proteinuria. While renal transplantation has cured SA in some patients, the post-transplant state is itself a risk factor for SA.

The high prevalence of SA in kidney disease and the associated clinical implications warrant vigilance in diagnosis and treatment of SA in the CKD patient. This review focuses on the prevalence of SA in patients with CKD including dialysis and transplant patients, and those with early-stage CKD and proteinuria. SA may vary in form and aetiology depending on type or stage of CKD. Based on these associations, we discuss our rationale for recommendations on screening and management of SA specific to the CKD population. The relationship between sleep apnoea (SA) and kidney disease represents a complex interaction of two disease processes where hypertension may or may not be an intermediary variable (Fig. 1). SA Resminostat has been associated with several types of renal disease, including proteinuria, early- and late-stage chronic kidney disease (CKD), end-stage renal disease (ESRD) on dialysis and renal transplantation. It is unknown whether there is a causative association in either direction between CKD and SA, or whether the two diseases represent clinical sequelae from a more common disease process, such as diabetes mellitus, hypertension or neuropathy. Given the complexity and variety of renal disease associated with SA, there may be different grounds for each particular association.

On the other hand, expression of the M2 marker genes encoding Arg1, Ym1, Fizz1, MR and IL-13 was severely impaired in Jmjd3−/− BM macrophages cultivated in the presence of M-CSF to induce M2 polarization, indicating that Jmjd3 is critical for M2 marker gene expression in BM macrophages. Although M-CSF-induced BM macrophages or chitin-induced peritoneal macrophages showed severe defects in M2 macrophage BGB324 molecular weight marker expression in the absence of Jmjd3, Jmjd3−/− BM macrophages were capable of upregulating expression of genes representative of M2 macrophages in response to IL-4 stimulation. These findings indicate that Jmjd3-mediated H3K27 demethylation is dispensable for the generation

of M2 polarization in response to IL-4, and suggest that M2 macrophages should be further subcategorized depending on the requirement of Jmjd3. Jmjd3 contains an N-terminal tetratricopeptide repeat domain and the JmjC domain in the C-terminus 32–34. The expression of the C-terminal part of Jmjd3 containing the JmjC domain, but not its demethylase-defective mutant, was sufficient to rescue M2 marker expression in Jmjd3−/− BM

macrophages. Therefore, Jmjd3 functions as a demethylase to induce M2 macrophage polarization, although recent studies show PD0325901 a demethylase-independent role in controlling chromatin remodeling together with T-box family transcription factors 40. Chromatin immunoprecipitation-sequencing (ChIP-Seq) analysis revealed that, in general, trimethyl H3K27 was enriched in the promoter regions close to the transcription start sites in BM macrophages. H3K27 of M2 marker genes, such as Arg1, Ym1 and Mrc1, were not trimethylated in the presence RVX-208 or absence of Jmjd3, suggesting that the M2 marker genes are not directly controlled by Jmjd3 through histone modification. On the other hand, H3K27 trimethylation of transcription factors such as Irf4 and Cebpb was differentially regulated by wild-type and Jmjd3−/− macrophages. The expression of Irf4 was diminished in Jmjd3−/− macrophages, and its expression was restored

in a Jmjd3 demethylase-dependent manner. Indeed, Irf4−/− mice showed severe defects in M2 macrophage polarization in response to chitin administration and induction of BM macrophages in the presence of M-CSF. Although Jmjd3 also controls a set of transcription factors, Irf4 is one of the critical target genes responsible for controlling M2 macrophage polarization (Fig. 2). Differential involvement of IRF transcription factors can be important for M1 and M2 macrophage polarization. It was reported that IRF5 is involved in the differentiation of M1 macrophages, though it is currently unclear whether Irf5 is epigenetically controlled by histone modifications 41. Jmjd3 is specifically involved in M2 macrophage polarization without affecting M1, despite the fact that Jmjd3 is TLR-inducible in macrophages.

[8, 9] The compound PGE2 is an arachidonic acid-derived lipid mediator generated in abundance at sites of infection and inflammation as a result of the rapid up-regulation of cyclooxygenase-2 and microsomal PGE synthase-1 enzymes.[10] It is also an important hormonal regulator of reproduction that is generated in the uterus where it is involved in early and late processes ranging from implantation of the fertilized egg to parturition.[11]

PGE2 is a highly potent modulator of innate and adaptive immunity that influences cell behavior through the ligation of its four distinct G-protein-coupled E-prostanoid (EP) receptors, numbered EP1-4.[12, 13] Both EP2 and EP4 are potent immunoregulatory receptors that share the capacity to increase intracellular concentrations of cyclic adenosine monophosphate (cAMP) within seconds to minutes of PGE2 binding.[13, 14] PGE2-dependent increases in cAMP have been shown to impair the phagocytic ability of different macrophage Crizotinib mouse types against a range of pathogens,[15-18] and it can be suggested that such effects might have evolved to limit the extent of host inflammatory responses or trigger the resolution of inflammation. However, in clinical situations such as pregnancy and the puerperium, where local and systemic PGE2 levels are elevated for physiological reasons,[19-21] the immunosuppressive effects of PGE2 might be maladaptive, particularly when an opportunistic selleck screening library pathogen such as C. sordellii gains access

to the normally uninfected uterus (or surrounding soft tissue). The purpose of this study was to address the question of whether PGE2 and cAMP-signaling cascades could regulate the phagocytosis of C. sordellii by human macrophages

and to determine the involvement and click here relative importance of EP2 and EP4 receptors in such regulation. A better understanding of endogenous regulators of innate immunity will enhance efforts to develop better preventive and therapeutic options against reproductive tract infections. Phorbol-12-myristate-13-acetate (PMA)-differentiated THP-1 cells (a human macrophage-like cell line) were used in this study. These cells were obtained from the American Type Culture Collection (ATCC, TIB-202; Manassas, VA, USA) and cultured in RPMI 1640 (Invitrogen, Carlsbad, CA, USA) supplemented with 1% antibiotic-antimycotic (Invitrogen) and 10% charcoal-/dextran-treated fetal bovine serum (FBS; HyClone, Waltham, MA, USA), referred to as RPMI +/+. Cells were passaged every 2–4 days and were used through the 10th passage, at which time a new culture was started. THP-1 cells were matured into macrophages by culturing with 100 nm PMA (Sigma-Aldrich, St. Louis, MO, USA) in RPMI +/+ for 24 hr at 37°C with 5% CO2. Cells were detached from the flask with non-enzymatic cell dissociation solution (Sigma-Aldrich) and gentle scraping. Phorbol-12-myristate-13-acetate-activated THP-1 cells were used for all experiments presented here, unless otherwise noted.