Although it is not yet well understood how it is ultimately determined which of these processes will assume the upper hand in any given situation, a few themes have emerged. Tolerance-promoting effects of iNKT cells appear to be clearly favoured when there is a lack of inflammatory stimuli in the local milieu, or when the level of antigenic stimulation is low. In contrast, exposure to an initial strong antigenic stimulus or to cytokine-mediated costimulation can favour the pro-inflammatory effects of iNKT cells. Questions that remain to be resolved include why in some cases iNKT cells nevertheless seem to contravene these ‘rules’, for example, by promoting tolerance in situations where there is substantial

inflammatory immune activation (e.g. organ transplantation). Seliciclib mw Based on our current picture, one thing that is a reasonably safe bet is that gaining a handle on how iNKT cells mediate their contrasting effects will not only reveal novel insights into the workings of these remarkable lymphocytes, but will also produce new information on the biology of DCs and other myeloid APCs. The authors were supported by National Institutes

of Health (NIH) grants AI074940 and AI076707, and by the Pew Scholars in the Biomedical Sciences Program. “
“To evaluate the effects of the anti-inflammatory and anti-angiogenic roles of LXA4 LBH589 datasheet on endometriosis in mice. Endometriosis was induced in 40 mice and separated into two groups. LXA4 group was administered by LXA4 for 3 weeks. The endometriotic lesions were counted, measured, and identified by pathology. The presence of a panel of pro-inflammatory factors was assessed by real-time RT-PCR, and enzyme-linked immunoassay, the mRNA, protein levels of matrix metalloproteinase (MMPs), and vascular endothelial growth factor (VEGF) were determined by real-time RT-PCR and immunohistochemistry;

Mephenoxalone the activity of MMPs was evaluated by gelatin zymography. Treatment with LXA4 significantly inhibited endometriotic lesion development (13.58 ± 4.01 mm2 in LXA4 group and 23.20 ± 7.49 mm2, P = 0.0002), downregulated pro-inflammatory factors, suppressed the activity of MMP9, and reduced the VEGF levels associated with endometriosis in mice. LXA4 may inhibit the progression of endometriosis possibly by anti-inflammation and anti-angiogenesis. “
“Fab fragments (Fabs) maintain the ability to bind to specific antigens but lack effector functions due to the absence of the Fc portion. In the present study, we tested whether Fabs of an allergen-specific monoclonal antibody (mAb) were able to regulate asthmatic responses in mice. Asthmatic responses were induced in BALB/c mice by passive sensitization with anti-ovalbumin (OVA) polyclonal antibodies (pAbs) (day 0) and by active sensitization with OVA (days 0 and 14), followed by intratracheal (i.t.) challenge with OVA on day 1 and days 28, 29, 30 and 35. Fabs prepared by the digestion of an anti-OVA IgG1 (O1-10) mAb with papain were i.t.

From 383 pregnancies referred in 2000-2013, 75 patients were selectedstage 1 CKD, referred within the 14th gestational week, singleton deliveries, absence of diabetes, hypertension or nephrotic proteinuria at referral, BMI<30); 267 “low-risk” pregnancies, followed in the same setting, served as controls. Glomerular filtration rateGFR) was assessed by CKD-EPI and dichotomized at 120 mL/min. The odds for caesarean section, prematurity, need for

Neonatal Intensive Care UnitNICU) were assessed by univariate analysis and logistic regression. Risk for adverse pregnancy outcomes was not affected by hyperfiltrationunivariate https://www.selleckchem.com/products/VX-770.html OR GFR >=120 mL/min: Caesarean section 1.300.46-3.65); preterm delivery: 0.840.25-2.80)). In contrast, even in these cases with normal kidney function, stage 1 CKD was associated with prematurity17.3% vs 4.9% p=0.001), lower birth weight3027 ± 586 versus 3268 ± 500 p<0.001) need for NICU12% vs 1.1% p<0.001). In the multivariate

analysis, the risks were significantly increased by proteinuria and maternal age but not by GFR. In pregnant Stage 1 CKD patients, hyperfiltration was not associated with maternal-foetal outcomes, thus suggesting to focus attention on qualitative factors, eventually enhanced by age, as vascular stiffness, endothelial damage or oxidative stress. “
“Novartis is delighted to report on the second renal transplant cases program held in 2013. The program was initiated with the aim of fostering and sharing innovation, development and the highest standards in the understanding and CX-4945 clinical trial clinical management of renal transplantation in Australia. This initiative was developed as part of the Novartis commitment to encouraging interest and education in the practice of Transplant Nephrology. Entries for these awards were permitted from any RACP Nephrology

Advance Trainee currently working in Australia. The submitted case reports were judged by an independent panel of distinguished Transplant Nephrologists who selected the top seven case reports according to: Scientific interest DNA Damage inhibitor Use of clinical acumen Clear and concise presentation We are delighted to sponsor the publication of the top seven cases, as chosen by the Panel, to be published in no particular order in this supplement of Nephrology. Novartis looks forward to providing more innovative programs as part of its commitment to excellence in the practice and research within the field of transplantation. “
“A 46-year-old woman presented with acute anuric renal failure preceded by 2 weeks of dry cough, fevers, loin pain and 2 days of profuse vomiting. She had been anuric for 24 h with marked intravascular fluid overload on examination. Oliguria persisted for 2 weeks and she required haemodialysis support before renal recovery. The aetiology of the illness was unidentified. She denied the use of any regular medications and any intravenous drug use. On examination there was no evidence of any needle marks and no drug screen was collected during admission.

In sum, modulation of the balance between autoimmunity and immunoregulation, and thus subsequent induction or prevention of T1D, might rely on the dual function of the innate immune players involved in the disease. Depending on timing and whether β-cell antigens are present, TLR-mediated effects will differentially affect the Erlotinib development of autoimmunity. The opposing roles of infections in T1D, which also depend on timing and vary in terms of damage to β cells 2, may thus be accounted for by the capacity of viruses to differentially affect such innate immune factors depending on the context. For instance, TLR2 signaling, and subsequent activation of

APCs/T cells and production of inflammatory cytokines, may promote autoimmune processes when β-cell antigens are present, but also appear to counter autoimmunity by enhancing and invigorating CD4+CD25+ Tregs and conferring

DCs with tolerogenic properties. Previous work has shown that TLR2 signaling enhances the function of CD4+CD25+ Tregs 22 and regulates their expansion and activity 29, 30. TLR2 was proposed to control antimicrobial immunity by transiently limiting the function of natural Tregs (thus permitting T-cell immunity) while enhancing their number (thus participating in terminating it). Accordingly, we found that acute anti-LCMV immunity coincided INCB018424 price with ineffective activity of CD4+CD25+ Tregs (data not shown) but resulted in their increased frequency and function. TLR2 might thus act to regulate antiviral immunity, by enhancing the number and function of Tregs to control it,

but impairing these cells as long as the invading virus is present. Intriguingly, to date, there is no evidence that LCMV particles can bind to TLR2. But while TLR2 is responsible for sensing components from micro-organisms, it can also recognize molecular motifs from certain endogenous ligands. In this regard, the chaperone HSP60 was shown to enhance the function of CD4+CD25+ Tregs through TLR2 signaling 22. It is thus possible that viral infection triggers the release of molecules such as HSPs, which promote the direct enhancement of CD4+CD25+ Selleckchem Atezolizumab Tregs via TLR2. This might constitute a means to recognize and control potentially harmful immune processes through innate immunity. Such absence of antigenic specificity could enable control of immunity to infection not only by viruses but also bacteria or other pathogens. In particular, in the hygiene hypothesis it is proposed that a number of different infections in early life contribute to reduced susceptibility to T1D 46. The capacity of the immune system to control immunopathology independent of antigen may thus account for the ability of numerous infections or non-infectious pro-inflammatory agents to protect from T1D in experimental models for this disease 13.

The biofilms formed by four out of seven strong slime-producer strains, after a 24-h incubation, are reported in Fig. 7, in which the typical tridimensional shape of a mature biofilm

is clearly evident in all the observed samples. Further, for the weak slime-producer strains of C. difficile, and P. bivia (Fig. 8) as well as for the two isolated strains of C. fallax (data not shown), it was possible to obtain a moderate development of a biofilm community after 48–72 h. A number of papers have reported possible hypotheses on the mechanisms presumably involved in the clogging phenomenon of biliary endoprostheses (for a review, see Donelli et al., 2007). To address the issue of how a biofilm could reach such a thickness to significantly narrow the lumen of the stent, it must be remembered that the biofilm exopolysaccharide matrix engulfs VX809 a number of ‘foreign bodies’ of different sizes including proteins, microbial byproducts, amorphous

calcium bilirubinate and crystals of fatty acid calcium Selleck Rapamycin salts, as well as large-sized dietary fibers (Groen et al., 1987; Leung et al., 1988; Sung et al., 1993; Basoli et al., 1999; Di Rosa et al., 1999; van Berkel et al., 2005). Bile viscosity, which differs on the basis of a patient’s health status, is another parameter to be considered. According to Poiseuille’s law, if the bile viscosity increases, the maintenance of the same bile flow would require an increase in the inner stent diameter: it has been calculated that an increase of 0.2 mm in the inner stent diameter corresponds to a 300% increase in bile flow (Rey et al., 1985). In fact, the narrowing of the stent lumen, as a consequence of biofilm development, causes the slowing of bile flow, promoting both spontaneous and bacteria-driven bile salt precipitation. Thus, considering a mean bacterial

Olopatadine diameter of about 1 μm, a reduction of 0.2 mm in a 10-Fr polyethylene stent (inner diameter 2.4 mm) would correspond to a biofilm of 200 overlapping bacterial layers. However, as already mentioned, the actual thickness of each bacterial layer is expected to be much higher because of the continuous engulfment of large-sized ‘foreign bodies.’ Further, the additional thickness of the host protein conditioning film, layered on the polymeric stent surface and known to mediate microbial attachment via specific adhesins, must be considered. This model, based on the progressive reduction of the stent lumen as a consequence of the multispecies biofilm expected to develop in the peculiar luminal microenvironment of a biliary stent, can be considered, in our opinion, to be a reasonable way to approach the critical issue of stent clogging. Moreover, the accumulation of biliary sludge is thought to be a multifactorial process in which, other than microbial growth, slime production and biofilm formation, the activity of some bacterial enzymes is involved. It is known that β-glucuronidase, produced by E.

CD4 T-cell

responses to complex protein Ags are restricted to a limited number of determinants, a process defined as peptide immunodominance.22 Several studies indicate that peptide immunodominance can be altered by the vaccine delivery systems used. Using MalE as a model Ag, Leclerc and colleagues found that altering the vaccine-delivery systems changed the number of MalE epitopes recognized by CD4 T cells.23 Immunization with a recombinant Salmonella strain expressing MalE protein allowed the presentation of MalE CD4 T-cell epitopes that were silent after administration of selleck compound purified MalE protein Ag in CFA.23 In a recent study, Andersen and colleagues reported that altering the vaccine-delivery systems used for a tuberculosis subunit vaccine based on fusion proteins of two mycobacterial Ags – ESAT-6 and Ag85B – also changed peptide immunodominance.24 While a recombinant Ag85B/ESAT-6 protein vaccine in a liposomal adjuvant induced primarily a CD4 T-cell response directed to an immunodominant epitope located in Ag85B, an adenovirus vector expressing the same fusion protein induced a strong CD8 response predominantly targeted to an epitope located in ESAT-6. Importantly, only the adjuvanted protein vaccine Selleck Caspase inhibitor gave efficient protection against subsequent Mycobacterium tuberculosis infection.24 Altogether these studies suggest that the formulation used to deliver a protein Ag can determine the specificity of the CD4 T-cell responses and the vaccine efficacy.

Adjuvants can alter the specificity of

the CD4 T-cell response by revealing cryptic epitopes or changing peptide dominance, but can they impact the clonotypic composition of a CD4 T-cell response directed against a defined immunodominant epitope? We have recently investigated the ability of five different adjuvants [Alum, CFA, incomplete Freund’s Phospholipase D1 adjuvant (IFA), CpG oligodeoxynucleotides (TLR9 agonists) in saline buffer and MPL-based emulsion] to elicit CD4 T-cell responses against the pigeon cytochrome c (PCC) protein in B10.BR mice.25 CD4 T-cell responses to PCC are directed against a single immunodominant peptide consisting of amino acids 88–104 presented by I–Ek.26 CD4 T cells specific for this epitope predominantly express Vα11 and Vβ3 TCR variable regions with restricted CDR3 features.27,28 All five adjuvants examined promoted a PCC-specific CD4 T-cell response dominated by clones expressing restricted TCRs, but the clonotypes selected varied across the different formulations. Dispersible adjuvants using TLR agonists (CpG, MPL) focused CD4 T-cell responses towards high-affinity clonotypes expressing TCR with a marked bias toward a public Vβ3-Jβ1.2 rearrangement (SLNNANSDY or 5C.C7β chain) in their CDR3β, as depicted in Fig. 1a. By contrast, adjuvants forming Ag deposition at the site of injection (alum, IFA and CFA) selected a more diverse CD4 T-cell response that was characterized by an increased prevalence of lower affinity clonotypes expressing Vβ3-Jβ2.

5% versus 0%, P = 0.001). Body weight did not change significantly in the icodextrin group, but body weight in the control group increased from 63.3 ± 14.5 kg at baseline to 64.2 ± 14.2 kg

at day 5 (P = 0.0002) and 65.2 ± 14.1 kg at day 10 (P < 0.0001). Conclusion: As compared with glucose-based peritoneal dialysis solution, use of icodextrin achieved better ultrafiltration and fluid control during acute peritonitis complicating continuous ambulatory peritoneal dialysis, although we found no evidence of a worthwhile clinical benefit on peritonitis resolution. (ClinicalTrial.gov number, NCT0104446 [ClinicalTrial.gov].) SUGIURA TOSHIHIRO1, AKAGAKI FUYUKO1, KUBOTA KEIICH1, NAKAMORI AYA1, WADA AKIRA2 1Otemae PI3K inhibitor cancer Hospital, Japan; 2Osaka National Hospital, Japan Introduction: Recent studies have shown that renal resistive index (RI) reflects systemic vascular stiffness as well as renal arteriolosclerosis. While this fact makes it difficult to interpret the increase in RI, we have shown that high RI is an independent risk factor for worsening renal function and can estimate renal prognosis in CKD [Nephrol Dial Transplant 2009; 24: 2780–5, Clin Exp Nephrol 2011; 15: 114–20]. The purpose of the present study is to determine the relative risks with an increase in RI for progression of CKD. Methods: We

performed a 2-year follow-up study with an observational cohort of 429 CKD patients (GFR 45 ± 31 mL/min/1.73 m2, age 57 ± 17 years). The patients were examined by Doppler ultrasonography for RI [(peak-systolic velocity – end-diastolic selleck chemicals velocity) / peak-systolic velocity] to be calculated. Glomerular filtration rate (GFR; mL/min/1.73 m2) was estimated from serum creatinine (s-Cr) and age with the revised Japanese equation: 194 × s-Cr−1.094 × Age−0.287 (×0.739 for women).

Worsening renal function was defined as a decrease in GFR of at least 20 mL/min1.73 m2 or the need for long-term dialysis therapy until the end of the 2-year follow-up. Results: Among the 429 CKD patients, 107 patients presented with worsening renal function during the 2-year follow-up. When we divided the patients into P-type ATPase three groups by RI value of 0.70 and 0.80, Kaplan-Meier analysis showed that the event-free survival rates of worsening renal function at 24 months were 0.93, 0.70 and 0.35 in patients with RI ≤ 0.70, 0.7 < RI ≤ 0.80 and RI > 0.80 respectively (Log-rank test, P < 0.001, Fig. 1). Cox proportional-hazard analysis showed that the adjusted hazard ratio (HR) for worsening renal function was 4.54 [95% confidence interval (CI) 2.31–8.96, P < 0.001] and 2.81 [95% CI 1.48–5.35, P < 0.01] in patients with RI > 0.80 and 0.7 < RI ≤ 0.80 respectively, as compared with the patients with RI ≤ 0.70. HR was adjusted by the factors that could influence RI itself and/or renal outcome, namely, age, GFR, urinary protein excretion, systolic blood pressure, and use of renin-angiotensin system (RAS) inhibitors.

While HIV-1 has been reported to induce DC maturation [47,62], there is considerably more evidence to suggest that HIV-1 does not induce maturation [44,63–67]. Because one measure of DC maturation is the surface

expression of distinct surface molecules, we first determined if HIV-1 infection influences the cell surface phenotype of MDDC during the course of maturation. After incubation with PF-02341066 concentration HIV-1 for 24 h and 48 h of culture, there was no change in the expression of CD80, CD86, CD83, CD40, CCR7, MHC-I or MHC-II, indicating that HIV-1 itself was not capable of inducing DC maturation. There was, however, an increase in DC-SIGN expression following HIV-1 infection (Fig. 3a). After iMDDC were infected with HIV-1 and then stimulated to mature, they expressed lower levels of CCR7 and MHC-II than that observed in uninfected cells (Fig. 3b,c), suggesting that HIV-1 inhibits the full maturation of iMDDCs. Functional analysis.  Analyses were conducted as follows. 1. Endocytosis: while a phenotypic analysis of MDDC can be used to partially

identify the maturation status of an MDDC, determining the effects of HIV-1 on the functional character of MDDC over the course of maturation is required to elucidate a comprehensive picture of the effects of HIV-1 on MDDC maturation. One critical function of DC is the uptake of antigen from HDAC phosphorylation the periphery for processing and presentation in lymphoid organs [3]. After endocytosing antigens, immature DC undergo maturation and move from the anatomic periphery to secondary lymphoid organs where their role becomes that of antigen presentation and not uptake [3,68]. As a measure of endocytic activity, and therefore the maturation state

of MDDC, the effect of HIV-1 on dextran uptake was evaluated. As expected, maturation of uninfected iMDDC resulted in during a decrease in FITC–dextran uptake (Fig. 4a). While HIV infection had no impact on the ability of iMDDC to take up dextran (Fig. 4b), HIV-1 infection was associated with blunted down-regulation of endocytosis by iMDDC (Fig. 4c). HIV-1 infection therefore appeared to inhibit maturation reflected by the fact that HIV-1 infected DC partially retain their endocytic function. 2. Antigen presentation: a primary function of DC is the presentation of antigens to naive T cells in peripheral lymphoid tissue [3]. The effect of HIV-1 infection on the ability of MDDC to present antigen to autologous CD8+ T cells was determined by incubating HIV-1-infected MDDC with autologous PBMC in the presence of a CEF peptide pool, as described previously [69]. After culturing CEF peptide-pulsed iMDDC with autologous PBMCs for 7 days, CD8+ T cells proliferated as expected (Fig. 5). When iMDDC were infected with HIV-1, however, CD8+ T cell proliferation in response to the CEF peptide pool was not observed (Fig. 5), suggesting that HIV-1 infection of DC prevented or interfered with antigen presentation. 3.

Briefly, race has been shown to modify the association between bacterial vaginosis and incident STI.25 One study found that certain cytokine and chemokine single-nucleotide polymorphisms were associated with ethnicity among HIV-infected individuals. The authors hypothesized ALK inhibitor that heritable variations in certain of these loci may contribute

to the acquisition or progression of HIV infection.26 Further, the concept of race is a complicated one. The National Institutes of Health has historically used self-identified racial categories. Individual patients frequently do not self-identify with one of these categories and thus are classified as ‘other’. A newer technology uses single-nucleotide polymorphisms to create families of ethnic derivation called ancestry informative markers.27 These require obtaining biologic samples and laboratory work by a reputable facility so are not used frequently. However, if race is an important component of an individual HIV risk study, consideration

can be given to collection of more detailed ethnicity data. There exists a vast body of literature detailing the association between genital tract infections and HIV acquisition find more and transmission. Much recent work has focused on herpes simplex virus-2 (HSV2) given the ulcerative and inflammatory nature of the infection and the high prevalence of the infection. If having HSV2 impacts shedding of HIV and the risk of transmission, then curbing the

shedding caused by this infection alone might decrease the burden of HIV infection worldwide. Herpes simplex virus-2 has been shown to increase viral load of HIV in both plasma and the genital tract, independent of the level of immunodeficiency.28 The etiology of increased shedding of HIV in the presence of HSV appears to be immunologically mediated. Rebbapragada et al.29 termed the interaction between HSV2 and HIV-1 ‘negative mucosal synergy’. While HSV suppression appears to decrease the risk of shedding HIV among women already infected with HIV, it does not appear to protect against acquisition or transmission of HIV-1.30,31 Herpes simplex virus-2 is not the only infection that alters mucosal immune handling of HIV. A less noticed but still not highly prevalent virus that may impact on genital shedding of HIV is human cytomegalovirus (CMV). The prevalence of CMV varies by geographical location, but after infection, it establishes lifelong latency. It can reactivate or hosts can be re-infected. A group well known for their CMV expertise recently developed a cervical explant study of CMV and HIV co-infection. They found that HIV appeared to enhance CMV in co-infected tissues which produced inflammatory cytokines. This explant model may be a useful tool for future studies examining the impact of CMV on HIV expression and vice versa.32 Frequently encountered STI have also been implicated in altering mucosal immunity.

Serum NGF levels varied greatly. Serum

NGF concentrations in healthy humans are not normally distributed. About 10% of healthy people have relatively high NGF concentrations.68,69 We also noted the same findings of serum NGF levels in OAB patients, but not in normal controls. The high serum NGF levels in healthy humans in other studies might result from underlying systemic conditions that affect serum NGF levels. C-reactive protein (CRP) is a protein found in the blood, the levels of which rise in response to inflammation. CRP is synthesized by the liver in response to factors released by fat cells (adipocytes).70 Serum CRP level can be used as a nonspecific marker of systemic inflammation. Chronic prostatic inflammation has been hypothesized to be associated with the BAY 57-1293 manufacturer pathogenesis of benign prostatic hyperplasia. However, the association between histological prostatic inflammation and LUTS is relatively weak.71 Rohrmann et al.72 reported that men with serum CRP levels >0.30 mg/dL were more likely to show three or four symptoms

(i.e. nocturia, incomplete emptying, hesitancy, and weak stream) from the Third National check details Health and Nutrition Examination Survey (NHANES III). Another report using longitudinal data from the Olmsted County study73 showed that patients with higher serum CRP levels were approximately two times more likely to exhibit a rapid increase in storage LUTS and almost 2.5 times more likely to show a rapid decrease in peak flow rate. Kupelian et al.74 reported a significant association between serum CRP level and overall International Prostate Symptom Score (IPSS) in both men and women included in the Boston Area Community

Health (BACH) survey. We this website also reported the serum CRP levels are associated with residual urgency symptoms in patients with benign prostatic hyperplasia after medical treatment.75 In women, serum CRP was also found to elevate in OAB patients. CRP levels were significantly higher in women with OAB-wet than in those with bladder oversensitivity and in the normal control group. Women with voiding dysfunction also had a non-significantly higher CRP level. Further analysis revealed that body mass index and maximum flow rate were two independent factors influencing CRP levels. However, serum CRP level is not considered a suitable biomarker for discriminating female non-SUI LUTD. As patients with OAB may have frequent detrusor contractions during the storage phase, it is possible that sustained isometric detrusor contractions could result in increased muscle bulk and hence increased detrusor wall thickness (DWT) or bladder wall thickness (BWT). It has been hypothesized that DWT increases in patients with DO.

The samples were then examined click here by phase-contrast and fluorescence microscopy for the level of phagocytosis.

To determine the numbers of colony-forming units of engulfed S. aureus, macrophages incubated with bacteria (macrophages : bacteria = 1 : 500) for 30 min were washed to remove unengulfed bacteria and further incubated for 30 min. The macrophages were lysed with water 0 and 30 min after washing, and the lysates at serial dilutions were seeded on agar-solidified mannitol salt medium or Luria–Bertani medium, the latter of which contained tetracycline and was used for bacteria transformed with the pHY300PLK-based plasmid. The plates were incubated overnight at 37°, and the number of colonies (only selleck kinase inhibitor those surrounded by yellow rings in the mannitol salt medium) was determined and presented relative to that obtained at time 0 after washing. For the determination of superoxide production, macrophages maintained on coverslips in serum-free RPMI-1640 medium were incubated with unlabelled bacteria (macrophages : bacteria = 1 : 1000) at 37°, and the amount of superoxide

released into the culture medium was determined by a chemiluminescence reaction using Diogenes, as described previously.10 To determine the activity of α-N-acetylglucosaminidase, whole-cell lysates of peritoneal macrophages were incubated in a reaction mixture containing 4-methylumbelliferyl N-acetyl-α-d-glucosaminide (Sigma-Aldrich), and the level of cleaved substrates

was measured with a fluorometer, as described previously.25 HEK293 cells were transfected by the calcium/phosphate method overnight with a mixture of plasmid DNA including pELAM26 (a gift from Dr Douglas Golenbock at the University of Massachusetts, Worcester, MA), a reporter gene vector expressing firefly luciferase under the control of a promoter activated by NF-κB; pRL-TK (Promega Corp.), a control reporter constitutively expressing luciferase from Renilla reniformis (Promega Dual-Luciferase Reporter Assay System) used for the normalization of transfection efficiency; and mouse TLR2 cDNA in pDisplay (Invitrogen, Carlsbad, CA) (a gift from Dr Yoshiyuki Adachi at Coproporphyrinogen III oxidase Tokyo University of Pharmacy and Life Science, Tokyo, Japan).27 The cells were further cultured with fresh medium for 1 day and subsequently incubated with S. aureus for 2 hr, and the cell lysates were examined for the amounts of firefly luciferase and Renilla luciferase using the Dual Luciferase Assay kit. The ratio of firefly luciferase to Renilla luciferase was determined and considered to represent the level of NF-κB activation. Data are representative of at least three independent experiments (n = 2–3 in each experiment) that yielded similar results. Data from quantitative analyses are expressed as the mean ± standard deviations of the results from at least three independent experiments.