The characterization of both antisera was reported recently.26 An inhibitor of transcription of messenger RNA (mRNA), actinomycin-D and an inhibitor of protein synthesis, cycloheximide, were purchased from BioMol International, L.P. (Plymouth Meeting, PA). Extraction-free CGRP enzyme-linked immunosorbent assay (ELISA) Kits were purchased

from Bachem (Torrance, CA). RAW 264.7 macrophages were cultured and maintained in DMEM containing penicillin/streptomycin (1 : 200) and 10% heat-inactivated FBS in a 37° incubator with 5% CO2 and 95% air. Cells were seeded at the density of 3 × 105 to 5 × 105/ml. Passages of 5–20 were used for the treatments. Lipopolysaccharide (1–1000 μg/ml) was used to treat cells for 3, 6, 12, 24 and 48 hr. Neutralizing IL-1β antiserum (1 and 10 ng/ml), IL-6 antiserum (1 and 10 ng/ml), NGF receptor chimera (1·5 and 5 μg/ml), selective COX2 inhibitor NS-398 (10 and 20 μm), neutralizing antisera against MAPK Inhibitor Library cost NGF receptor trkA (1 : 1000), CLR antiserum (1 : 500 and 1 : 1000), RAMP1 (1 : 500 and 1 : 1000), PGE2 (1–30 μm), actinomycin-D (1 μm) and cycloheximide (1 μm) were learn more used alone or in co-treatment with LPS (1 μg/ml). The PGE2 and NS-398 were dissolved in ethanol and prepared as 10-mm stock solutions. Co-treatments lasted for 24 hr. Culture media were collected and stored at −80° until further

analysis. All treatments were performed in triplicate and each experiment was repeated at least three times. Following treatment, culture media were collected in pyrogen-free Eppendorf tubes and frozen at − 80° or underwent ELISA immediately. An extraction-free CGRP ELISA Kit was used. All procedures were performed according to the manufacturer’s instructions and the microplate was read using a microplate reader (Molecular Devices, Sunnyvale, CA). The detection range for CGRP was 0–10 ng/ml. Each treatment was performed in triplicate for each experiment. The mean value of CGRP released in culture medium following

Amine dehydrogenase treatments was compared statistically among groups. The RAW 264.7 macrophages were maintained in DMEM containing penicillin/streptomycin (1 : 200) and 10% FBS. Cells were seeded at a density of 3 × 106 to 5 × 106/ml in 24-well culture plates. Passages of 5–20 were used for the following treatments. Vehicle, LPS (1 μg/ml), CGRP (1, 10 and 100 nm), CGRP8-37 (0·1, 1 and 10 μm) and BIBN4096BS (0·01, 0·1 and 1 μm) were used to treat cells for 24 hr. Culture media were collected and stored at − 80°. All samples were assayed for MCP-1, IL-1β, IL-6, TNFα and IL-10 according to the manufacturer`s instructions using Mouse Cytokine Lincoplex Kits (Linco Diagnostic Services Inc., St Charles, MO). Each treatment was repeated at least three times. The mean and SEM were determined for each treatment and compared statistically among groups. Each treatment was performed in triplicate in each session of experiments.

44 It was shown that the double knockout mice had an even greater increase in B1-cell expansion, while the B2 population showed a reduction in size.44 Neither CD22 nor siglec-G single knockout mice showed development of autoimmunity whereas aged CD22,

siglec-G double knockout mice showed spontaneous development of anti-DNA autoantibodies and displayed a mild form of immune complex Lumacaftor manufacturer glomerulonephritis.44 These data suggest that CD22 and siglec-G may have par-tial overlap in the regulation of B-cell signalling and tolerance. The negative regulatory role of CD22 on B cells is well characterized but whether siglecs play a role in inducing tolerance in immune cells had not been explored until recently. Duong et al.45 showed that decoration of TI-2 antigens with sialic acids induces poor immune responses and leads to tolerance. Both siglec-G and CD22 have been shown to play a role in inducing tolerance,

preventing plasma cell differentiation and survival.45 This is the first report of tolerance being induced through siglecs in addition to their established role in dysregulation of Selleck HSP inhibitor cell signalling. Host response to injury is a relatively neglected component of innate immunity that is often viewed simply as a system that discriminates between self and non-self. Matzinger first proposed the ‘danger theory’ in 1994, in which she argued that rather than differentiating between self and non-self, the immune system discriminates between dangerous and non-dangerous signals, whether it is from an external or internal source.46 Like pathogen-associated www.selleck.co.jp/products/sorafenib.html molecular patterns (PAMPs), which interact with TLRs to stimulate immune response against pathogens, danger-associated molecular patterns (DAMPs) are released during injury and are thought also to bind TLRs and induce an inflammatory response.47 The DAMPs include heat-shock protein 70, heat-shock protein 90, high mobility group box

1 (HMGB1) and cellular RNA.47,48 Using a paracetamol-induced liver necrosis model, CD24, a glycosylphosphatidylinositol-anchored protein, has been identified as a receptor that interacts with the danger signal, HMGB1 and acts to protect against paracetamol-induced hepatotoxicity.48 CD24-deficient mice showed strong pro-inflammatory responses to paracetamol treatment: increase in IL-6, monocyte chemotactic protein-1 and TNF-α.48 Liver damage was indicated by an increase in serum alanine transaminase, indicative of liver haemorrhage and necrosis.48 Siglec-10 was shown to bind to CD24 and proposed to transduce inhibitory signalling that protects the mice against a lethal response to liver cell death.48 This was supported in studies of siglec-G (mouse orthologue of siglec-10) deficient mice which also showed greater inflammatory responses to high-dose paracetamol injections.48 The response of dendritic cells cultured from wild-type, CD24−/− and siglec-G−/− mice to the DAMP signal HMGB1 was compared with the PAMP signal LPS.

Using the same gating strategy as in Fig. 1A, a small population of Lin− Thy1+ Sca1+ ILCs could consistently be detected in healthy WT animals (Fig. 1D). To exclude artifacts resulting from a potential inadvertent inclusion of T cells, we also analyzed Rag1−/− mice, which completely lack T and B cells, as well as TCRβδ−/− mice, which lack all T cells. Indeed, we could verify that the CNS of healthy Rag1−/− as well

as TCRβδ−/− mice also contained a population of Lin− Thy1+ Sca1+ cells. SB203580 chemical structure IL-7R-α expression was detectable irrespective of the analyzed genotype (Fig. 1D). Quantification showed that the amount of ILCs in the CNS during steady state conditions, both in absolute numbers as well as in percentage, was similar in WT, Rag−/− and TCRβδ−/− animals (Fig. 1E). Due to their lack of lineage

markers and their rarity, their precise location within the uninflamed CNS is thus far unclear. In contrast to the steady state, a drastic increase this website in ILCs was observed under inflammatory conditions (Fig. 1E), suggesting that Thy1+ Sca1+ ILCs infiltrate into or expand in the CNS during experimental autoimmunity. In order to obtain a more detailed view on the temporal expansion of ILCs, we analyzed the CNS of MOG/CFA-immunized animals at different time points postimmunization, namely on day 8 (prior to disease onset), day 13 (peak disease), and day 18 (postpeak disease). While prior to disease onset very few Thy1+ Sca1+ ILCs could be detected, the number of ILCs on days 13 and 18 postimmunization was comparable. However, ILCs numbers vary at later disease time points, potentially correlating with the extent of remission from the disease. One of the most prominently studied features of RORγt+ ILCs is their immediate responsiveness to IL-23 and their ability to produce proinflammatory cytokines,

including IL-17 [3], IL-22 [10], and also IFN-γ [11]. In innate intestinal inflammation, both IL-17 and IFN-γ produced by ILCs have been shown to greatly contribute to disease progression [11]. Therefore, Mannose-binding protein-associated serine protease we analyzed cytokine production of CNS-infiltrating ILCs ex vivo by intracellular cytokine staining and found that a large population of Thy1+ Sca1+ ILCs was able to produce IFN-γ, and to a lesser extent IL-17 (Fig. 2A). We could not detect any expression of IL-22 (data not shown). Analysis of cytokine expression by CNS-resident ILCs during steady state showed only minor production of both IFN-γ and IL-17 (Fig. 2B). Since PMA/ionomycin is a very strong activator, we asked whether cytokine production by Thy1+ Sca1+ ILCs could be directly induced by stimulation with IL-23. Indeed, in vitro culture in the presence of IL-23 induced IL-17 production by CNS-isolated ILCs comparable to the levels observed with PMA/ionocycin (Fig. 2C).

In addition, those who responded may have been more motivated to respond because they had differing practices that they wanted expressed to the immunology community anonymously, or they actually are well versed in practice guidelines and wanted to portray this fact by responding to our survey. Those who did not respond may have differed in their comfort level in caring for immunodeficient patients or believed that they had nothing novel to contribute by responding. Given that clinical immunology is sometimes a separate subspeciality within parts of Europe, the majority of those who received the questionnaire should have been equally comfortable in caring for

PID patients with a similar find more familiarity in practice guidelines, so this bias would be expected to be minimal. This might have explained the small but measurable

difference in response rate between ESID and the AAAAI. IVIg is well documented to decrease infection rates within Metformin mouse specific PIDs [7,8]. The recommendation of IVIg as therapy for patients with PID varies with specific disease and there was agreement between ESID and focused AAAAI respondents in most diagnoses (Fig. 1). For example, all three subgroups agreed in their recommendation of IVIg for X-linked agammaglobulinaemia. For common variable immune deficiency (CVID), 96·9% of ESID respondents recommended treating most to all patients with IVIg compared with 90·5% of general AAAAI respondents, although this difference was not statistically significant (P = 0·057). Hyper-IgM (HIGM) syndrome presented a more dramatic difference, where 92·9% of ESID respondents recommended use of IVIg to treat the majority of these patients, whereas only 51% of general AAAAI respondents agreed (P < 0·001). These differences were not apparent when ESID and focused AAAAI respondents were compared (Fig. 1). In addition, ESID respondents recommended Pyruvate dehydrogenase lipoamide kinase isozyme 1 IVIg more frequently than general

AAAAI respondents for severe combined immune deficiency (SCID) (P < 0·001), whereas the responses of the focused AAAAI respondents were statistically indistinguishable from those of ESID. The differences were largely the same as those identified previously between the general and focused AAAAI members [5]. These findings are likely to indicate a need for increased awareness of practice parameters and guidelines for the treatment of PID among subspecialists who divide their effort among immunology and other disciplines, as well as increased education in PID. A substantial proportion of general AAAAI members practice in a community-based setting that further distinguishes this group from ESID, and creates a potentially unique set of educational needs and challenges. There are complex PID diseases where guidelines are less clear regarding use of IVIg therapy [9], and in these cases responses varied more within the experienced groups.

The association of loss of FUBP1 protein expression and either 1p/19q LOH or IDH-1 mutation was analysed using the likelihood-ratio Chi-square test. A significance level of alpha = 0.05 was selected for all tests. The sensitivity was calculated by dividing the number of genetically CCI-779 confirmed mutated cases by the number of FUBP1-negative cases as assessed by immunohistochemical analyses in the cohort of genetically tested samples.

The specificity was calculated by dividing the number of genetically confirmed nonmutated cases by the number of FUBP1-positive cases in immunohistochemical analysis. Statistical analysis was performed using JMP 8.0 software (SAS, Cary, NC, USA). Evaluation of the immunohistochemical preparations and photographic documentation was performed using an Olympus MI-503 cost BX50 light microscope. We first screened normal CNS tissue to examine the cellular distribution of FUBP1 protein under nonpathological conditions. In the cortex, neuronal nuclei exhibited strong FUBP1 expression, while intermingled glial or endothelial cells were negative or displayed only very weak FUBP1 expression

(Figure S2A). Moreover, normal white matter displayed only single cells with weak to moderate FUBP1 expression levels and FUBP1 signals were almost completely absent in oligodendrocytes constituting the largest white matter cell Progesterone population (Figure S2B). NIH REMBRANDT database analyses revealed significantly elevated FUBP1 mRNA expression levels in human glial neoplasms as compared with normal CNS specimens (URL: https://caintegrator.nci.nih.gov/rembrandt/legal.jsp) (Figure S3). However, no significant differences in the FUBP1 expression profile were observed between the various glioma subtypes. We next examined whether this increase in FUBP1 mRNA correlated with FUBP1 protein levels in glial neoplasms. Most cases of oligodendrogliomas (Figure 1),

astrocytomas and glioblastomas (Figure 2) displayed a strong increase in FUBP1 protein expression as compared with normal glial cells (Figure S2B). To analyse whether FUBP1 protein expression is associated with markers currently assessed in routine neuropathological diagnostics, we further examined the expression levels of FUBP1 (Figures 1A,E,I,M,2A,E,I), mutated IDH1 (R132H) (Figures 1B,F,J,N,2B,F,J), the MIB-1 index (Ki-67) (Figures 1C,G,K,O,2C,G,K) and p53 (Figures 1D,H,L,P,2D,H,L) in glioma subtypes. The median FUBP1 expression score was comparable for all glioma subtypes with WHO grade II oligodendrogliomas showing the lowest median expression score (median score, 7; range, 0–12).

aeruginosa and S. aureus grown in a flow-chamber system. We demonstrated how adaptive mutations in regulator genes of P. aeruginosa affect interactions between P. aeruginosa and S. aureus in co-culture biofilms. Pseudomonas aeruginosa

wild-type PAO1 (Holloway & Morgan, 1986), P. aeruginosa mucA mutant (Hentzer et al., 2001), AZD4547 clinical trial P. aeruginosa rpoN mutant (Webb et al., 2003), P. aeruginosa pilA mutant (Klausen et al., 2003b), P. aeruginosa pilH mutant (Barken et al., 2008), P. aeruginosa pqsA mutant (D’Argenio et al., 2002), S. aureus MN8 (Yarwood et al., 2004), S. aureus ISP479 (Toledo-Arana et al., 2005) and S. aureus 15981 (Toledo-Arana et al., 2005) were kindly provided by the cited authors and used in the present study. The pDA2 plasmid (An et al., 2006) was used DZNeP to complement the pilA mutant. Fluorescence-tagged strains were constructed by the insertion of a mini-Tn7-eGFP-Gmr cassette as described (Koch et al., 2001; Klausen et al., 2003b). Escherichia coli strains MT102 and DH5α were used for standard DNA manipulations. Luria–Bertani medium (Bertani, 1951) was used to cultivate E. coli strains. A modified FAB medium (Qin et al., 2007) supplemented with 0.3 mM glucose and 3% of Tryptic Soy Broth (TSB, BD Diagnostics) was used for biofilm cultivation. Selective media were supplemented with ampicillin (100 mg L−1), gentamicin (60 mg L−1) or carbenicillin

(200 mg L−1). Biofilms were grown in flow chambers

with individual channel dimensions of 1 × 4 × 40 mm at 37 °C. The flow system was assembled and prepared as described previously (Sternberg & Tolker-Nielsen, 2006). Overnight cultures of P. aeruginosa and S. aureus were diluted to an OD600 nm of 0.001. The flow chambers were inoculated by injecting 350 μL of monospecies diluted cultures or P. aeruginosa–S. aureus 1 : 1 mixed-species diluted cultures into each flow channel with a small syringe. After inoculation, flow channels were left without flow for 1 h, after which medium flow (0.2 mm s−1) was started using a Watson Marlow 205S peristaltic pump. For DNase I treatment, biofilm medium was supplemented with 20 μg mL−1 bovine DNase I (Sigma) from the beginning of cultivation. All microscopic observations and image acquisitions were performed using a Zeiss LSM 510 confocal laser scanning microscope (Carl Zeiss, Jena, Galeterone Germany) equipped with detectors and filter sets for monitoring of green and red fluorescence from general nucleic acid staining SYTO 9 (Invitrogen) and gram-positive specific staining hexidium iodide (Invitrogen) (Mason et al., 1998), respectively. BacLite Live/Dead viability stain (Molecular Probes, Eugene, OR) was used to visualize dead and live cells in co-culture biofilms. Images were obtained using a × 40/1.3 objective. Simulated three-dimensional images and sections were generated using the imaris software package (Bitplane AG, Zürich, Switzerland).

“
“Aims:  Low estimated glomerular filtration rate (eGFR) is associated with high mortality after stroke. However, ageing can influence eGFR directly and limit this burden impact. We investigated if low eGFR can be a predictor of death in different age groups after ischaemic stroke. Methods:  We evaluated and followed for 22 ± 14 months 871 unselected consecutive survivor patients more than 30 days after ischaemic stroke (55%

men, mean age of 66 ± 13 years) recruited in a prospective Brazilian cohort study from March 2005 to December 2007. Traditional cardiovascular risk factors and eGFR by The Chronic Kidney Disease Epidemiology Collaboration formula were analyzed as predictors of mortality for the whole cohort population and stratified by age (younger or older than 65 years old) in a Cox proportional hazards regression model. Results:  There were 119 (14%) deaths during follow up. The mean eGFR Selleckchem Daporinad was 74 ± 23 mL/min per 1.73 m2. Three hundred and sixteen patients (36%) presented eGFR lower than 60 mL/min per 1.73 m2. For the whole population,

eGFR lower than 60 mL/min per 1.73 m2 was independently associated with death after stroke in the multivariate analysis. When stratified by age groups, low eGFR was the single and independent predictor of death just for individuals younger than 65 years-old, as for older people just chronic atrial fibrillation, previous stroke and increase of age were associated with death. Conclusion:  Low eGFR measured at the first day of Cabozantinib supplier hospital admission can be a simple and trustful predictor of death after ischaemic stroke in people younger than 65 years old. “
“Aim:  Hepatic ischaemia/reperfusion injury (IRI) frequently complicates acute kidney injury (AKI) during the perioperative period. This study was to determine whether

hepatic IRI causes AKI and the effect of the sphingosine-1-phosphate (S1P) on AKI. Methods:  S1P and vehicle were given to mice before ischaemia and mice were subjected to hepatic IRI. Plasma creatinine (PCr), Selleck Temsirolimus alanine transaminase (ALT), urinary neutrophil gelatinase-associated lipocalin (NGAL) and renal histological changes were determined. As a marker of endothelial injury, vascular permeability was measured. The effect of VPC 23019, a S1P1 receptor antagonist, was also assessed. Results:  Hepatic IRI resulted in liver injury (increased ALT) and systemic inflammation. Kidneys showed elevated inflammatory cytokines, leucocyte infiltration, increased vascular permeability, tubular cell apoptosis and increased urinary NGAL, although PCr did not increase. Pretreatment with S1P resulted in an attenuation of systemic inflammation and kidney injury without any effect on plasma ALT or peripheral lymphocytes. The protective effect of S1P was partially reversed by VPC 23019, suggesting the important contribution of the S1P/S1P1 pathway to protect against hepatic IRI-induced AKI.

, 2009; Stübs et al., 2009), and the antigenic nature of ACGal has been confirmed by chemical synthesis (Stübs et al., 2010). These data imply that ACGal could improve serodiagnostics,

and may act as a basis for vaccine development. However, to date, it is unclear whether detection of or vaccination with ACGal would encompass LD-causing genospecies other than B. burgdorferi click here sensu stricto, B. afzelii, and B. garinii. On the other hand, the function of ACGal in B. burgdorferi is not elucidated, and the report that acylated cholesteryl α-d-glucosides in Helicobacter pylori are associated with immune evasion (Wunder et al., 2006) raises the question of whether ACGal are involved in the pathogenesis of LD. Therefore, in this study, we wanted to determine whether ACGal is a feature of other genospecies STI571 of B. burgdorferi sensu lato, including those associated with all stages of LD as well as B. spielmanii as an agent of localized LD. The following Borrelia strains were grown under microaerophilic conditions in 9 mL of BSK-H medium at 33 °C as described previously (Preac-Mursic et al., 1986): B. burgdorferi s.s.

strain B31, B. afzelii PKo, B. bavariensis PBi, B. garinii A and TN, B. spielmanii PSig II, B. bissettii DN 127, B. lusitaniae Poti B2 and Poti B3, B. valaisiana VS 116 and UK, B. japonica HO 14, B. hermsii HS 1. The methods and materials for harvesting and extraction of bacteria have been described in detail earlier. In brief, the cells were harvested, lyophilized, and disintegrated using an ultrasonic rod and the lipids were extracted by a Folch extraction (Folch et al., 1957). The total lipids were dissolved and spotted in about equal amounts on a thin-layer chromatogram (TLC). Synthetic ACGal was applied as a reference (Stübs Carbohydrate et al., 2010). The chromatography was performed in chloroform/methanol 85 : 15 v/v.

The lipids were visualized on the TLC by molybdenum stain. The dried TLC was immersed in buffer and blotted onto a polyvinylidene difluoride (PVDF) membrane using a hot iron. The membrane was blocked with a skim milk/phosphate-buffered saline solution and incubated for 13 h at 4 °C with a 1 : 750 diluted serum of LD patients in the late stage. The membrane was incubated for 1.5 h at room temperature with a 1 : 50 000 dilution of a secondary, horseradish peroxidase-conjugated anti-human IgG antibody. The serum antibody binding was detected using enzymatic chemoluminescence to expose and subsequently develop X-ray films. Dot blots and Borrelia lysates were generated as described previously (Stübs et al., 2010): ACGal, Borrelia lysate and total lipids were spotted on PVDF membranes and incubated with pooled sera (n=4) from patients diagnosed with LD, syphilis as well as leptospirosis at 4 °C for 15 h. Detection with secondary antibodies was performed via chemoluminescence. The stained TLC (Fig. 1a) revealed that all analyzed Borrelia genospecies exhibited a similar lipid pattern.

The principle aim of this study was to analyze the number of capb copies, and to assess sequence divergence in the hcsA and hcsB genes of Hib strains isolated from

children with Hib diseases in our district before the introduction of the Hib conjugate vaccine. A total of 24 Hib strains isolated between November 2004 and May 2009 from 24 children with invasive Hib diseases who had not received Hib conjugate vaccine in Kagoshima Prefecture, Japan, were collected and examined. Of these strains, 15 were isolated from CSF and 9 from blood. The strains were epidemiologically unrelated and individually stored at −80°C. All isolates were identified as serotype b by PCR capsular genotyping (14). PFGE was performed using a CHEF-DR 3 apparatus (Nippon Bio-Rad Laboratories, Tokyo, Japan) according to previously reported methodology (15). Briefly, DNA was digested by SmaI and separated on 1% agarose gels by PFGE under the following

learn more conditions: current range, 100 to 130 mA at 14°C for 16 hr; initial switch time, 5.3 s, linearly increasing to a final switch time of 49.9 s; angle, 120°; field strength, 6 volts/cm. The gels were stained with ethidium bromide and photographed. A lambda with a size range of 48.5 kb to 1 Mb (BME, Rockland, ME, USA) was used as a size marker. For interpretation of banding patterns separated by PFGE, we referred to the criteria of Tenover et al. (16). Fulvestrant concentration Two variants of the capb locus DNA sequence, type I and type II, were determined by PCR using two primer sets targeting the hcsA gene which could discriminate between the two capsular genotypes as described in a previous report (12). The DNA sequences of the PCR products were determined Thymidylate synthase by an ABI Prism 310 sequencer (Applied Biosystems Japan, Tokyo, Japan). The number of capb locus copies was detected by Southern blotting analysis according to previously reported methods (8). Because KpnI and SmaI restriction sites flank the capb locus, extracted DNA in an agarose plug was digested with these enzymes, separated by PFGE, and transferred to a nylon membrane. A Hib

capsule-specific 480-bp probe was constructed by PCR (14) and labeled with DIG using a DIG high prime DNA labeling kit (Roche Diagnostics, Mannheim, Germany). The membrane was hybridized with the probe and visualized by chemiluminescent detection using a DIG detection kit (Roche Diagnostics). The Kpn I/Sma I fragment of a two copy strain was expected to be 45-kb, because it includes two repeats of the locus (18 + 17 kb) plus additional segments (∼10 kb) upstream and downstream of the cap region (17). Three-, four-, and five-copy fragments showed increased size in 18-kb increments for each additional copy (63, 81, and 99-kb, respectively) (8). A summary of results is shown in Table 1. The type I-associated hcsA gene was found in all of the strains examined. The DNA sequences of all the PCR products were completely identical. PFGE analysis showed nine distinctive restriction patterns (A to I) among the 24 isolates.

, 2005a). In contrast, heat-inactivated P. acanthamoebae elicited several cytokines (IL-6, TNF-α, 12p40) (Roger et al., 2010). Chlamydia trachomatis can elicit cytokines in the live and inactivated form, but the level and kind of cytokines are not necessarily the same (O’Connell et al., 2006; Schrader et al., 2007; Bas et al., 2008). If Chlamydia muridarum, a mouse Fer-1 in vivo pneumonitis strain adapted to be a model for C. trachomatis urogenital infection, was heat-inactivated or treated with UV, the expression of certain

cytokines, such as IL-1β, was absent (Prantner et al., 2009) or decreased, such as TNF-α and IL-6 (Darville et al., 2003). Chlamydia pneumoniae also required to be viable to induce IL-6, IL-12 and TNF-α production (Geng et al., 2000). Therefore, depending on the species, some antigens are not effective anymore if exposed to heat or UV denaturation. In contrast, other antigens present on the bacterial surface may be resistant to heat (such

as lipids) and therefore still be able to induce cytokine expression. Depending on the cytokines, bacterial growth and protein synthesis might be required. Moreover, the kind of macrophages and the stimuli used to induce macrophage differentiation probably influence the cytokine expression pattern. A priming of the macrophages with lipopolysaccharides or other PAMPs yielded a much higher production of IL-1β upon C. muridarum infection (Prantner et al., 2009). Previous exposure of macrophages to antigens Meloxicam or RBs from lysed epithelial cells could therefore allow a much stronger and rapid response to chlamydial infection. Not all the Chlamydiales seem to have the

same susceptibility to cytokines. Some are restricted TGF-beta inhibitor in their growth while others can circumvent them or even use them to their advantage (Haranaga et al., 2003; Jendro et al., 2004). Expression of cytokines upon chlamydial infection was, to some extent, confirmed in animal models (Table 2). The role of innate and adaptive immunity in clearance and disease progression of C. trachomatis has been reviewed recently (Miyairi et al., 2010; Rank & Whittum-Hudson, 2010). Because non-human primate studies have only been investigated with C. trachomatis, we will not discuss them in this minireview. Chlamydia muridarum infection caused an upregulation of cytokines, such as IFN-γ, IL-6, IL-1β and TNF-α, and a whole range of chemokines as well as cytokine/chemokine receptor expressions (Rank et al., 2010). Cytokine knockout mice are a powerful tool to assess the role of cytokines in bacterial clearance and pathogenesis. So far, this has been performed to a small extent, for example in C. muridarum infections in IL-12 or IL-18 knockouts (Lu et al., 2000b) and IL-10 knockouts for C. pneumoniae (Penttiläet al., 2008), but should be extended to other members of the Chlamydiales order. Lung infection with C. muridarum was severely increased in IL-12 knockout mice, while the absence of IL-18 did not significantly affect clearance of the bacteria (Lu et al.