2D). However, similar reduction was observed in TNF-α secretion (Fig. 2E), suggesting that the slight reduction in IL-1β secretion in Pkr−/− macrophages is not related to inflammasome activation. Our initial studies were performed with

PKR-deficient mice in which the N-terminal RNA binding domain of PKR was deleted [17]. In contrast, Lu et al. studied PKR using KO mice with deletion of the catalytic domain of PKR [18]. Although both KO mice lack expression of full-length PKR, some conflicting results have been reported for these two mouse mutant strains [19]. Therefore, we also studied inflammasome activation in macrophages from mutant mice with deletion of the catalytic domain of PKR. Analysis of macrophages Vismodegib nmr from this Pkr−/− mouse strain also revealed comparable caspase-1 activation and pro-IL-1β/IL-18 processing in response to activators of the NLRP3 inflammasome when compared LY294002 order with that of WT macrophages (Fig. 3A). As expected, caspase-1 activation

and pro-IL-1β/IL-18 procession were abrogated in macrophages from Nlrp3−/− mice (Fig. 3A). Likewise, caspase-1 activation and pro-IL-1β maturation induced by aluminum salts (Alum), another activator of NLRP3, were unimpaired in Pkr−/− macrophages, but abolished in Nlrp3−/‒ macrophages (Fig. 3B). 2-aminopurin (2-AP), a potent inhibitor of PKR, was reported Glutathione peroxidase to inhibit ATP-induced NLRP3 inflammasome activation at millimolar concentration [8]. Notably, addition of 2-AP at this high concentration inhibited ATP-induced NLRP3 inflammasome activation in both WT and PKR-deficient macrophages (Fig. 3C). This result suggests at this high concentration, 2-AP inhibits the inflammasome through off-target effects. Furthermore, caspase-1 activation in response to Salmonella or poly (dA:dT) were unaffected by deletion of the catalytic domain of PKR (Fig. 3D and E). Consistent with these results, IL-1β and TNF-α release induced by ATP, Salmonella and poly (dA:dT) were unimpaired in Pkr−/− macrophages (Fig. 3F and G). Our results indicate that the protein

kinase PKR plays a critical role in regulating iNOS production by macrophages after LPS challenge, which correlated with reduced intracellular killing of E. coli. However, we found no detectable role for PKR in the activation of the NLRP3, NLRC4 or AIM2 inflammasomes in macrophages. We do not have a clear explanation for the difference in results between our studies and those of Lu et al. [8]. It is possible that subtle variation in experimental conditions may account, at least in part, for the differences in results. In our studies, parallel experiments were performed using macrophages from mice deficient in NLRP3 and NLRC4 that showed requirement for these inflammasomes, but not PKR, for caspase-1 activation triggered by specific stimuli.

In the case of percentage change, the change should be calculated as 100 × [(final – original)/(original)] values. Most of the time, we wish to summarize a set of measurements and also report a comparison. Let’s look at our jumping frogs [3]. We reported the mean distances that these groups jumped, to the nearest AG-014699 chemical structure millimetre. Perhaps that was a little optimistic, as at the competition the jump length was measured to the nearest ¼ inch! How shall we summarize the results of that first test on trained and untrained frogs? To describe the first sample we studied (Figure 1) we need

to express two different concepts to characterize the samples. We give a measure of central tendency to summarize the magnitude of the variable (examples would be the mean or median values) and a measure of dispersion or variability (such as a standard deviation or quartiles). In the case of our frogs, the jump lengths and variation of the jump lengths are the important features of our sample. These distances were normally distributed (not a common feature in most biological experiments) so to describe the overall performance of the untrained frogs we report the sample mean distance jumped. To describe the variability or spread,

we report the sample standard deviation. The final value needed to characterize the sample is the number of measurements. So, in the case of the untrained frogs, we report that the distance jumped was 4912 (473) mm, BTK inhibitor clinical trial where these values are mean (SD), and we could add (n = 20) if we have not already stated the size of the sample used. These values summarize our estimate of the population characteristics. There is no added benefit in using

the symbol ± and reporting 4912 ± 473 mm. In fact the use of this convention can be confusing: is it the mean ± SD, the mean ± SEM (defined shortly) or a confidence Sitaxentan interval? Many authors choose to use the standard error of the mean (SEM) as a measure of the variability when describing samples. This is incorrect: this value should only be used to indicate the precision with which the mean value has been estimated. As we saw in our frog studies, this value depends very much on the sample size. We told our readers how many frogs we sampled, which is how we achieve the precision. Note that care should be taken when interpreting the SEM as it stands. Here, it is the standard deviation of the sampling distribution of the mean. It tells us how the sample mean varies if repeated samples of the same type (with same sample size) were collected and the mean calculated. In fact 68% of these estimated means would be expected to lie in a range between one SEM less and one SEM greater than the actual population mean. Thus there remain a substantial proportion of sample means that do not fall within this range. To assess how precisely a sample mean has been characterized, the preferred measure of precision of a mean estimate is the 95% confidence interval.

59 The menstrual regularity was maintained and women continued to have ovulatory cycles.60 No change in

bleeding profile was observed. With the approval of the Drugs Controller General of India and Institutional Ethics Committees, phase II efficacy trials were carried out with this vaccine in three major institutions: the All India Institute of Medical Sciences (AIIMS, New Delhi), Postgraduate Institute of Medical Education and Research (PGIMER, Chandigarh), and Safdarjung Hospital, New Delhi. A total of 148 sexually active women of proven fertility with two living children (of which one below 1 year to confirm their contemporary fertility) https://www.selleckchem.com/products/poziotinib-hm781-36b.html were enrolled with their informed consent. Many of them had come to clinics earlier for medical termination of unwanted pregnancy. The available contraceptives in the family planning basket either did not

suit these women or were not used consistently. Their husbands were reluctant to use condoms. Primary immunization was given by three intramuscular injections of the HSD-TT/DT vaccine adsorbed on alum at monthly interval. Sodium phthalyl lipopolysaccharide (SPLPS), a non-pyrogenic derivative of LPS, was used at 1 mg in the first injection only. Vaccine with the TT or DT as carrier was given alternatively, selleck chemical so as to avoid carrier-induced suppression of antibody response to HSD. All women made antibodies reactive with hCG.4 However, 110 of the 148 immunized women had hCG bioneutralization titers above 50 ng/mL (a threshold fixed for testing protection against pregnancy) for 3 months or longer. All women continued to ovulate and had regular menstrual cycles. The antibody titers declined with time but booster injections raised the titers (Fig. 4). Eight women completed more than 30 cycles by voluntary intake of booster injections as and when required without becoming pregnant. Nine completed 24–29 cycles, 12 completed 18–23 cycles, 15 completed 12–17 cycles, and 21 women completed 6–11

cycles. The personal diary of women indicated without doubt that they were sexually active with a minimum of two sexual intercourses per week. The semen parameters of husbands were good with high counts of motile sperms. The fact that the women were prone to become pregnant Fossariinae is supported by the record of 26 pregnancies taking place in women at titers falling below 35 ng/mL bioneutralization capacity. Fig. 5 is an illustrative example of a 30-year-old subject with two living children and one MTP. After three primary injections of the vaccine, she took two boosters and remained protected against pregnancy for 13 cycles. In the immediate cycle, when her antibody titers had fallen below 20 ng/mL, she conceived and had a positive pregnancy test. Although most conceptions occurring at or below protective threshold were terminated at the behest of the subjects (Medical termination of pregnancy is legal in India), four women decided to continue with their pregnancy.

In addition to documenting the safety of this

approach, we found that patients treated with OK432-stimulated DCs displayed unique cytokine and chemokine HDAC inhibitor drugs profiles and, most importantly, experienced prolonged recurrence-free survival. Inclusion criteria were a radiological diagnosis of primary HCC by computed tomography (CT) angiography, hepatitis C virus (HCV)-related HCC, a Karnofsky score of ≥ 70%, an age of ≥ 20 years, informed consent and the following normal baseline haematological parameters (within 1 week before DC administration): haemoglobin ≥ 8·5 g/dl; white cell count ≥ 2000/µl; platelet count ≥ 50 000/µl; creatinine < 1·5 mg/dl and liver damage A or B [23]. Exclusion criteria included severe cardiac, renal, pulmonary, haematological or other

systemic disease associated with a discontinuation risk; human immunodeficiency virus (HIV) infection; prior history of other malignancies; history of surgery, chemotherapy or radiation therapy within 4 weeks; immunological disorders including splenectomy and radiation to the spleen; corticosteroid or anti-histamine therapy; current lactation; pregnancy; history of organ transplantation; or difficulty in follow-up. Thirteen patients (four women and nine men) presenting at Kanazawa DAPT solubility dmso University Hospital between March 2004 and June 2006 were enrolled into the study, with an age range from 56 to Reverse transcriptase 83 years (Table 1). Patients with verified radiological diagnoses of HCC stage II or more were eligible and enrolled in this study. In addition, a group of 22 historical controls (nine women and 13 men) treated with TAE without DC administration between July 2000 and September 2007 was included in this study. All patients received RFA therapy to increase the locoregional effects 1 week later [24]. They underwent ultrasound, computed tomography (CT) scan or magnetic resonance imaging (MRI) of the abdomen about 1 month after treatment and at a minimum of

once every 3 months thereafter, and tumour recurrences were followed for up to 360 days. The Institutional Review Board reviewed and approved the study protocol. This study complied with ethical standards outlined in the Declaration of Helsinki. Adverse events were monitored for 1 month after the DC infusion in terms of fever, vomiting, abdominal pain, encephalopathy, myalgia, ascites, gastrointestinal disorder, bleeding, hepatic abscess and autoimmune diseases. DCs were generated from blood monocyte precursors, as reported previously [25]. Briefly, peripheral blood mononuclear cells (PBMCs) were isolated by centrifugation in LymphoprepTM Tubes (Nycomed, Roskilde, Denmark). For generating DCs, PBMCs were plated in six-well tissue culture dishes (Costar, Cambridge, MA, USA) at 1·4 × 107 cells in 2 ml per well and allowed to adhere to plastic for 2 h.

8,9 Similarly, in humans, correlative data suggest that Crohn’s disease is driven by exaggerated Th1

and Th17 responses, because inflamed lesions contain increased levels of Th1-associated and Th17-associated Bcl-2 inhibitor cytokines including interferon-γ, IL-12, IL-17 and IL-18.23–27 In contrast, although ulcerative colitis is in the same family of diseases, it is associated with a Th2 cell profile, and patients have high levels of IL-13 in the mucosa compared with Crohn’s disease patients or healthy controls.19,23,28 Hence, although in most cases T-cell dysfunction is unlikely to be the initiating cause of IBD,29 there is substantial evidence that dysregulated Th cell responses perpetuate the disease and the vicious cycle of chronic inflammation. Under normal conditions, compared LDK378 price with all other tissues, the intestinal lamina propria has the greatest proportion of CD4+ Tregs,30 which are thought to be primarily specific for antigens in food and commensal flora.29 As Crohn’s disease and ulcerative colitis are both T-cell-driven diseases, it logically follows that increasing appropriate Treg activity in the gut should help to restore the balance of suppression

in inflamed tissues. However, it is unknown whether the over-abundance of activated T cells in IBD is the result of a numerical lack of Tregs, a defect in their function, resistance of T effector cells to suppression, or a combination of these possibilities. These questions have not been widely studied in animal models, yet they

are key to understanding whether restoring/boosting Tregs is likely to have any effect in treating IBD in humans. There is evidence that simply lacking Tregs leads to IBD. Patients with genetic mutations in FoxP3 who have non-functional or absent Tregs always have severe intestinal inflammation associated with lymphocytic infiltration of the intestinal mucosa.31,32 Similarly, mice lacking Protein kinase N1 FoxP3+ Tregs,33 or the ability to suppress via Treg-derived cytokines such as IL-10,34,35 IL-35,36 and in some cases TGF-β,37 develop severe colitis. In the more common forms of IBD, however, there is little evidence to suggest that patients simply lack Tregs in the circulation and/or the affected tissues. Maul et al.38 found that although both Crohn’s disease and ulcerative colitis patients had decreased Treg populations in the peripheral blood during active disease, Treg numbers in intestinal tissue biopsies were not substantially different from those in patients with other inflammatory diseases. Other studies corroborate these results, and in most cases show a consistent expansion of Tregs in both inflamed and non-inflamed sections of the gut in adult and paediatric patients with IBD.

The superiority of IL-10−/− DC for vaccine delivery

is thus well explained immunologically by their improved abilities to provide both the antigen-specific and essential co-stimulatory signals 68, and to reach rapidly the secondary JQ1 solubility dmso lymphoid organs where adaptive immune responses are initiated. The findings are also in agreement with several previous studies on the role of suppressor of cytokine signalling (SOCS) molecules in regulating DC immunogenicity. SOCS are a group of intracellular negative regulators of JAK/STAT signalling, and the expression of some of its members (SOCS1 and SOCS3) is also associated with IL-10 receptor triggering 70. The SOCS1 molecule, for example, is a potent suppressor of DC and macrophage activation 71–73. DC from SOCS1-deficient mice are hyper-responsive in vitro, and spontaneously activated in vivo. Interestingly, SOCS1−/− mice also develop a spontaneous lupus-like disease MK-8669 nmr indicating a crucial role of this molecule in regulating self-reactivity 71. Most importantly, it has been demonstrated that the inhibition of SOCS1 could enhance significantly the abilities of DC to present tumour antigens, to produce IL-12 and to induce effectively anti-tumour responses 73–75. The lack of IL-10 could therefore

potentially render DC resistant to the tolerogenic tumour microenvironment, hence to the conversion of “regulatory” or “tolerogenic” DC 38. This may have further impact on DC functions by alleviating certain inhibitory signals through other negative receptors expressed on DC. DC-derived Ig receptor

2 (DIgR2) is, for example, an inhibitory receptor associated with immunoreceptor tyrosine-based inhibitory motifs (ITIM), which could be up-regulated on DC in response to IL-10. It has recently been demonstrated that selective blocking DIgR2 on DC could enhance their immunogenicity in Janus kinase (JAK) vitro, and tumour vaccines delivered by the DIgR2-silenced DC elicited potent anti-tumour immune responses in vivo in mouse models 76. In conclusion, emerging evidence indicates that one of the most effective ways to enhance the efficacy of DC-based tumour immunotherapy is by targeting the negative arm of immune regulation. The removal of DC-IL-10, in particular, breaks directly and effectively the negative feedback loop thus alleviating the immunosuppressive impacts of tumours on the host immune system. It allows the generation of immunologically optimised DC vectors, which can provide potentially both strong antigen-specific triggers and essential co-stimulatory signals, for inducing tumour-specific immunity even under the highly immunosuppressive tumourigenic microenvironment (Fig. 1).

Importantly, anti-tumour monoclonal antibodies (mAbs) or bispecific Abs (BsAbs) —

which link Fc receptors on immune cells and tumour-associated antigens (TAAs) on tumour cells — enhance neutrophil-mediated tumour cell lysis [8-10]. Initially, the immunoglobulin (Ig) G receptor FcγRI was proposed as a potent target for initiation of neutrophil-induced Ab-mediated tumour cell lysis. In recent years, however, it was demonstrated STI571 purchase that targeting the IgA Fc receptor (FcαRI) resulted in more effective neutrophil-mediated Ab-dependent tumour cell lysis [11-19]. Furthermore, killing was initiated through non-apoptotic pathways, which coincided with autophagic characteristics [20]. Moreover, triggering of FcαRI induced recruitment of click here neutrophils into tumour colonies [9]. We recently demonstrated that IgA induced significant release of the neutrophil chemoattractant leukotriene B4 (LTB4) [21]. Thus, neutrophils represent interesting effector cells for Ab immunotherapy of cancer. However, in order to achieve Ab-mediated lysis of solid tumours in vivo, neutrophils need to extravasate from the circulation into the tumour. Therefore, we now investigated Ab-induced neutrophil migration towards tumour colonies in the presence of an endothelial cell barrier. Neutrophils were previously

demonstrated to induce Ab-dependent killing, which resulted in tumour cell elimination [8, 9, 11-13, 16, 17, 19, 22]. Moreover, FcαRI proved a more potent trigger molecule, as compared buy Osimertinib with targeting FcγRs [9, 13, 15]. Interestingly, we recently demonstrated that cross-linking of neutrophil FcαRI by IgA resulted in release of LTB4, which is a potent neutrophil chemoattractant [21]. As such, rapid migration of neutrophils was observed towards the site of the IgA-immune complexes. Similarly, when we added an FcαRIxHer-2/neu BsAb to a 3D culture of tumour cells in collagen, we observed massive neutrophil migration towards tumour colonies within 2 h (Fig. 1A). At

this time point only minimal degranulation was observed (reflected by lactoferrin release, Fig. 1B). However, neutrophil degranulation increased over time in cultures in which FcαRIxHer-2/neu BsAb had been added. We previously showed in a 2D culture system that incubation of SK-BR-3 cells and neutrophils in the presence of an FcαRIxHer-2/neu BsAb resulted in tumour cell death [20]. Although we formally cannot show tumour cell killing in our 3D collagen cultures, the integrity of tumour colonies was clearly affected after 24 h incubation with neutrophils and FcαRIxHer-2/neu BsAb (Fig. 1A, panel VI; inset). Chemotactic activity was only observed in the supernatants of cultures in which FcαRIxHer-2/neu BsAb had been added, which was decreased in the presence of a blocking anti-BLTR1 mAb (Fig. 1C and D). This suggested that the observed rapid neutrophil migration was the result of LTB4 release after triggering of FcαRI [21]. Additionally, release of the pro-inflammatory cytokines IL-1β and TNF-α was observed (Fig.

Virulence factors such as elastase and protease have been proposed to play an

important role in the Ixazomib initial establishment of lung infections (Elsheikh et al., 1987; Smith et al., 2006b) and are also important in acute infections, such as keratitis (Wilcox et al., 2008). These virulence factors have as well been shown to be present at elevated levels during acute exacerbations in patients with CF (Grimwood et al., 1993; Jaffar-Bandjee et al., 1995). In contrast, reduced expression of these virulence factors is associated with chronic CF isolates (Smith et al., 2006a; Tingpej et al., 2007; Bjarnsholt et al., 2010). It was observed here that the STY variants of strain 18A showed a dramatic increase in elastase activity and thus appear to regain hallmarks of acute infection isolates. This suggests that the expression of such virulence factors and the switch between acute and chronic infection types may be a reversible process. Moreover, strain 18A variants showed an increase in the production of acylated homoserine lactones, further showing that the loss of such phenotypes

by chronic infection isolates is reversible. The ability of P. aeruginosa to convert back to an acute infection phenotype may also explain the development of acute exacerbations during lung infection in patients with CF (Grimwood et al., 1993; Jaffar-Bandjee et al., 1995). The morphotypic and phenotypic variants studied here were stable, indicating that the morphotypic conversion was linked to a mutation. To better understand the processes driving the development of these variants, the mutation frequencies of selleck the parental strains were investigated and the 18A parent strain was shown to have a 3.4-fold higher mutation frequency than strain PAO1. The lower mutation frequency observed for PAO1 may reflect differences in long-term selection, based on laboratory cultivation on defined media, compared to the lung environment with constant exposure to the host immune response, antibiotic challenge and invading strains. The mutation frequencies were determined at multiple stages of biofilm development for both strain Lepirudin 18A and strain PAO1, and it was observed that they fluctuated approximately

10- and 4.5-fold, respectively. The mutation frequencies also correlated with the growth phases of the biofilms (Fig. 5 and data not shown) with a decline in the mutation frequency when the biofilm biomass was increasing. Interestingly, Garcia-Castillo et al. (2011) have reported that biofilm populations of CF isolates showed a lower mutation frequency compared to planktonic cultures. In contrast, Conibear et al. (2009) demonstrated a 100-fold increase in mutation in biofilm cells, specifically within microcolonies, compared to planktonic cells. The biofilm populations studied [(our study) vs. subpopulations (Conibear et al., 2009)] could account for the differing findings. To explore further this generation of diversity, a number of genes that might contribute were subsequently sequenced.

Sensory nerves could play a role in the transient vasodilation, which

is less well understood [71]. Such transient vasodilation is more obvious when the cooling is rapid [147], making the rate of cooling an important parameter to consider when studying microvascular reactivity to local cooling. We recently assessed the reproducibility of skin blood flux measurements while cooling locally to 15°C or to 24°C on the forearm. Pembrolizumab The best seven-day reproducibility of a 30-minute cooling protocol was obtained at 15°C when data were expressed as percentage decrease from baseline flux (CV = 23%) [116]. This test has been recently used to characterize increased vasoconstriction and blunted vasodilation on the finger of patients with primary RP compared with matched controls [115]. LSCI is a recently marketed technique based on speckle contrast analysis that provides

an index of blood flow [12,50]. High frame rate LSCI allows continuous assessment of skin perfusion over wide areas, thus theoretically combining the advantages of LDF and LDI, with very good inter-day reproducibility of PORH and LTH measurements, whether data are expressed as raw values or as a function of baseline [117]. It should be noted that the skin penetration see more depth of LSCI is about 300 μm, whereas it is deeper (about 1–1.5 mm) with laser Doppler techniques [11,106]. There are little data about the linearity between the LSCI signal and actual skin blood flow in human skin, whereas LDI has been shown to provide a valid measure of skin blood flow [49,76]. Recent work based on computer simulations and laboratory measurements has shown that LDI and LSCI similarly provide a perfusion index proportional to the concentration and mean velocity of red PAK5 blood cells [131]. In vivo, Stewart et al. have shown a very good correlation between the

two techniques in burn scar perfusion assessment [127]. Such correlation between LSCI and LDI is maintained over a wide range of human skin perfusion when data are expressed as raw arbitrary perfusion units [98] (Figure 7). Subtracting BZ from raw arbitrary perfusion units did not affect the correlation between LSCI and LDI, but shifted the regression line toward the origin [98]. A potential problem of LSCI is its sensitivity to movement artifacts. Mahe et al. recently showed that movement-induced artifacts may be overcome by subtracting the signal backscattered from an opaque adhesive surface adjacent to the ROI [90]. This simple method could be useful in many investigations of skin microvascular function when strict immobility cannot be ensured. Analyzing LSCI is challenging, partly because of the large amount of data (i.e., an acquisition rate of 18 Hz provides more than 40,000 images for a single 40-minute LTH measurement). Rousseau et al.

The strongest response was induced by peptide 10–26 followed by peptides 289–306, 117–133/120–133, and 46–70, as determined by high levels of IFN-γ as well as the presence of IL-2 in culture supernatants (Fig. 2). The two peptides 117–133 and 120–133 led to a similar IFN-γ response, although the longer sequence click here induced significantly more IL-2 (p=0.009). In addition, peptide 46–70 stimulated the production of higher amounts of IFN-γ and IL-2 compared to peptide 50–70 (Fig. 2), showing the importance of flanking residues for the induction of an optimal T-cell response. Of note, peptide 305–322, indicated as good binder to DR*0401 by TEPITOPE

(Table 1), did not bind in our assay (Fig. 1) nor did it induce a T-cell response in DR*0401-Tg mice (Fig. 2). Therefore, this peptide was not selected for analysis in RA patients. In conclusion, the four best binders to DR*0401, as determined by binding assays and TEPITOPE program (Table 1), were also the most

immunogenic ones in DR*0401-Tg mice (Fig. 2 and Table 1). We next assessed the potential of the selected peptides to induce production of IFN-γ in PBMC from RA patients and healthy individuals. Freshly isolated PBMC from 33 RA patients and 16 healthy controls were cultured with 13 individual hnRNP-A2 peptides (indicated in bold in Table www.selleckchem.com/products/abt-199.html 1) in ELISPOT plates pre-coated with an anti-IFN-γ mAb. In this assay, PBMC from 6 out of 33 (18%) patients showed an IFN-γ response to hnRNP-A2 peptides, five of them (15%) to a main determinant contained in peptide 117–133 (Fig. 3 and Table 2). The mean frequency of IFN-γ-producing cells specific for this dominant epitope was 21±9 out of 106 cells (mean/duplicate for each patient: 25, 15, 11, 20, 39, 15) compared to 2±2 out of 106 cells (3, 0, 0, 4, 5, 0) Histidine ammonia-lyase for the medium background. Remarkably, when retesting two of the six reactive patients 3 months after the first evaluation, the T-cell response to the peptide was sustained (Fig. 3 and Table 2). Conversely, PBMC from none of the healthy individuals reacted to hnRNP-A2

peptides (Table 2). Of note, T-cell reactivity to hnRNP-A2 peptides was independent of disease duration, which varied between 3 and 14 years, and immunosuppressive medication (Table 2 and Supporting Information Table 1). Importantly, all six patients with peptide reactivity presented with active disease (DAS28 > 3.2), and four out of five had bone erosions. We next thought to confirm these findings, to show that the responses to peptides 117/120–133 are mediated by CD4+ T cells, and to investigate whether they are selectively found in RA patients. To demonstrate MHC class II restriction, we incubated the cells with an anti-class II Ab together with peptides 117/120–133 and analyzed the proliferative response in 25 additional RA and 28 disease control (DC) patients with osteoarthritis (Supporting Information Table 2).