Metabolic syndrome has become a major public health challenge worldwide. Patients with MS are twice as likely to have cardiovascular disease and four times as likely to have type II diabetes mellitus than patients without MS. The mortality rate of cardiovascular or coronary arterial diseases is also increased 2.9–4.0 times in patients with MS.1–3 Recently, the incidence of obesity has risen rapidly, even in

Asian countries, making obesity-related disease a paramount concern.4,5 MS is one of the most important obesity-related phenotype complexes of hypertension, insulin resistance, low high-density lipoprotein (HDL)-cholesterol and hypertriglyceridemia.6 There is increasing evidence from clinical and epidemiological studies of associations between lower urinary tract symptoms (LUTS) and major chronic medical diseases as well as related lifestyle factors.7 These associations have stimulated interest in the contribution of factors R428 outside the urinary tract to urological symptoms; the so-called “beyond the bladder” hypothesis.8,9 Traditionally, the pathogenesis of benign prostatic Fulvestrant mw hyperplasia (BPH) and LUTS was explained as interaction between hormonal

and genetic factors. Increasing clinical significance of BPH and LUTS with increasing age is not only a urological issue. With increasing life expectancy, people demand better health-related life quality, which could cause an obesity epidemic. Several cohort studies have independently reported that components of MS, including obesity, influence the development of BPH and LUTS. Increasing evidence suggests that modifiable risk factors may participate in the development of BPH and LUTS. The characteristics of MS may differ due to a population’s socioeconomic and cultural basis. Metabolic syndrome is defined based on the Adult Treatment Panel III of the National Cholesterol Education Program (ATPIII NCEP) diagnosis criteria as three or more of the following cardiovascular risk factors: (i) abdominal obesity (waist circumference ≥88 cm); (ii) elevated blood pressure

(≥130 mmHg systolic or ≥85 mmHg diastolic); (iii) high triglyceride level (≥150 mg/dL); (iv) glucose intolerance (fasting glucose Anacetrapib ≥110 mg/dL); and (v) low HDL-cholesterol level (<50 mg/dL).10 In the present study, body mass index (BMI) ≥25 kg/m2 was used as an indicator of obesity instead of waist circumference.11 For Asian populations, BMI ≥25 kg/m2 the new cut-off value for obesity because Asians have a proportionally higher percentage of total body fat and abdominal fat than Caucasians with the same BMI and thus obesity-related complications occur at a lower BMI.12–14 It is important to apply unified definition criteria, but racial and cultural differences should be considered among different populations in different countries. In a study of Asian North Indians, modified NCEP ATP III criteria showed the highest occurrence of MS in incident-type II diabetes mellitus (DM) patients.

Soluble CD23 is also found in the saliva of Sjögren’s syndrome

patients41,42 and in the plasma of patients with systemic lupus erythematosus,41,42 though in the case of systemic lupus erythematosus the effect of sCD23 is likely to be mediated via its interaction with CD21 on autoimmune B cells rather than via integrins on monocytic cells.43 The finding of high sCD23 levels in such syndromes has made both sCD23 protein itself and its various receptors attractive targets for therapeutic intervention. This aspiration is supported by data from rodent systems where anti-CD23 mAbs have been shown to both prevent initial and ameliorate existing NVP-BKM120 clinical trial arthritic disease,25,26 and by the success of Lumiliximab, a humanized macaque anti-CD23 antibody, in treatment of B chronic lymphocytic leukaemia,44 a disease characterized by strikingly high plasma sCD23 levels.45 A different strategy, employing a CD23-binding peptide identified by phage display technology, also shows promise in preventing onset of adjuvant-induced arthritis

and reducing severity of established disease in rats.46 The identification of αVβ3 as an sCD23 receptor linked to TNF-α release in human monocytes18 suggested that antibodies to this integrin might be useful in autoimmune inflammatory disease.47 The Etaracizumab Dorsomorphin mAb (Abergrin, Vitaxin),48,49 a humanized form of the LM609 anti-αVβ3 reagent, was shown to be potent in inhibiting angiogenesis.50,51 However, Etaracizumab was also assessed in psoriatic arthritis but was not found to have a therapeutic effect and this is potentially explained by the fact that the parent LM609 mAb does not inhibit sCD23-driven TNF-α release from monocytes,18 a finding that implies that the mAb does not influence the site on the integrin responsible for control of cytokine release. Our data that showed LM609 did not induce cytokine production from either THP-1 or U937 cells (Fig. 3) were also in agreement with this

suggestion. Etaracizumab retains significant Vasopressin Receptor promise, however, and is currently in trials for therapy of metastatic melanoma.52 It is important to bear in mind that most previous studies on integrin function have been performed in adherent cells. The possibility of an alternative mode of integrin signalling illustrated by sCD23 is particularly interesting in the context of haematopoietic cells, including monocytes, which are non-adherent cells, but nonetheless express a wide range of integrins, and are the precursors of a number of adherent, terminally differentiated cells, such as macrophages and osteoclasts. The differentiation of monocytes into adherent counterparts is the result of paracrine or autocrine signalling in response to cytokines, such as those released by the interaction of sCD23 with integrins.

Here, we investigated the effects of VEGF on selleck chemicals sciatic nerve regeneration. Methods: Using light and electron microscopy, we evaluated sciatic nerve regeneration after

transection and VEGF gene therapy. We examined the survival of the neurones in the dorsal root ganglia and in lumbar 4 segment of spinal cord. We also evaluated the functional recovery using the sciatic functional index and gastrocnemius muscle weight. In addition, we evaluated the VEGF expression by immunohistochemistry. Results: Fluorescein isothiocyanate-dextran (FITC-dextran) fluorescence of nerves and muscles revealed intense staining in the VEGF-treated group. Quantitative analysis showed that the numbers of myelinated fibres and blood vessels were significantly higher in VEGF-treated animals. VEGF also Selleckchem Tamoxifen increased the survival of neurone cell bodies in dorsal root ganglia and in spinal cord. The sciatic functional index and gastrocnemius muscle weight reached significantly higher values in VEGF-treated animals. Conclusion: We demonstrate a positive relationship between increased vascularization and enhanced nerve regeneration, indicating that VEGF administration can support and enhance the growth of regenerating nerve fibres, probably through a combination of angiogenic, neurotrophic

and neuroprotective effects. “
“The antiphospholipid syndrome (APS) is an autoimmune disease characterized by high titers of auto-antibodies (aPL) leading to thrombosis and consequent infarcts. However, many affected patients develop neurological symptoms in the absence of stroke. Similarly, in a mouse model of this disease (eAPS), animals consistently develop behavioral abnormalities despite lack of ischemic brain injury. Therefore, very the present study was designed to identify structural alterations of hippocampal neurons underlying the neurological symptoms in eAPS. Adult female Balb/C mice were subjected to either induction of eAPS by immunization with ß2-Glycoprotein 1 or to a control group.

After sixteen weeks animals underwent behavioral and cognitive testing using Staircase test (experiment 1 and 2) and Y-maze alternation test (experiment 1) and were tested for serum aPL levels (both experiments). Animals of experiment 1 (n=7/group) were used for hippocampal neuron analysis using Golgi-Cox staining. Animals of experiment 2 (n=7/group) were used to analyse molecular markers of total dendritic integrity (MAP2), presynaptic plasticity (synaptobrevin 2/VAMP2) and dendritic spines (synaptopodin) using immunohistochemistry. eAPS mice developed increased aPL titers and presented with abnormal behavior and impaired short term memory. Further, they revealed a reduction of dendritic complexity of hippocampal CA1 neurons as reflected by decreased dendritic length, arborization and spine density, respectively. Additional decrease of the spine-associated protein expression of Synaptopodin points to dendritic spines as major targets in the pathological process.

Thus, IL-7 must be controlling naïve T-cell survival by mechanisms other than simply regulating expression level of Bcl2 family members. Taken together, our data

strongly suggest that IL-7 controls homeostatic fitness of T cells in replete hosts by non-transcriptional mechanisms. IL-7 can activate PI3K 23, 37 and downstream Akt/PKB whose kinase activity can potentially modulate multiple pathways and that could constitute such non-transcriptional mechanisms. Consistent with this view, IL-7 has been reported to prevent apoptosis in IL-7 responsive cell lines by inhibiting Bad activity following Y-27632 order Akt/PKB phosphorylation of Bad 31. However, using F5 T cells over-expressing Bad, we could find no evidence that Bad was regulating naïve T-cell fitness in vitro, or in vivo in a range of homeostatic environments or in the absence of IL-7 signalling altogether. This is also consistent with experiments showing that inhibition of PI3K does not block the

pro-survival properties LDK378 of IL-7 23 in vitro. However, in vitro, any potential pro-apoptotic consequence of PI3K blockade may be masked by the effects of upregulation of Bcl2 expression by IL-7. Furthermore, it is unclear whether or to what extent IL-7 activates PI3K in naïve T cells in vivo. Thus, it is not possible to exclude a potential pro-survival role for IL-7-dependent PI3K activation in vivo. The non-transcriptional mechanisms by which IL-7 promotes T-cell survival in vivo remain obscure. However, since we observed no differences in abundance of key Bcl2 family members in IL-7R− F5 T cells, it seems likely that regulation at the level of sub-cellular localization of pro- or

anti-apoptotic proteins and/or their interaction with one another may rather account for the perturbed mitochondrial homeostasis we observed in IL-7R− F5 T cells. Furthermore, another study suggests that posttranslational regulation of glucose transporters may be involved 36. In conclusion, we show for the first time that homeostatic fitness of T cells is dynamically regulated by IL-7, involving multiple mechanisms that differ between lymphoreplete and lymphopenic conditions. Bacterial neuraminidase The view that T-cell fitness is not a digital state of either survival or death but rather dynamic state is consistent with concepts of competition for survival resources. Such a view is also consistent with the recent insights into the high mobility of lymphocytes within the 3-dimensional structure of the lymph node 38, 39, and that the source of IL-7, and likely other survival factors within these structures, is not homogeneously distributed, but rather focal and from specific cell types 11. In such a context, a dynamic fitness model of T-cell survival would permit integration and interpretation of multiple and likely sporadic survival cues.

The membrane was incubated with primary antibody and an appropriate secondary horseradish peroxidase-conjugated antibody. Signals were detected by enhanced chemiluminescence (GE Healthcare Bio-sciences, Little Chalfont, UK). The immunoreactive Vemurafenib supplier bands were scanned to produce digital images that were quantified employing SCION Image software, and fold phosphorylation was calculated from the amount of phospho-protein relative to the corresponding non-phospho loading control. The IgE-sensitized cells (1×106) were loaded with 4 μM Fluo3-AM (Dojindo, Kumamoto, Japan) for 30 min at 37°C. The cells were resuspended in 1×Tyrode’s

buffer, and then changes in dye fluorescence upon the addition of stimulants were monitored employing flow cytometry. [Ca2+]i mobilization was expressed as the relative fluorescence intensity. Data shown are the mean±SD. Statistical analysis was performed using Student’s t-test. Probability values <0.05 were considered to indicate statistically significant differences. This work was supported by the grants-in-Aid for private universities from the Ministry of Education, Culture, Sports, Science (C. Ra), and Technology of Japan, the

Grants-in-Aid for Scientific Research from the Nihon University (C. Ra). Conflict of interest: The authors declare no financial or commercial conflict of interest. “
“Although data show Palbociclib solubility dmso the importance of type I interferons (IFNs) in the regulation of the innate and adaptive immunity elicited in response to viral, bacterial and parasitic infections, the functional activities of these cytokines during fungal infections are poorly

understood. We examined here the impact of IFN-β on the response of human monocyte-derived dendritic cells (DCs) infected in vitro with Aspergillus fumigatus. Having found that A. fumigatus-infected DCs do not express IFN-β, we evaluated the effect of the exogenous addition of IFN-β on the maturation of human DCs induced by the infection with A. fumigatus conidia. Although the phagocytosis of the fungus was not affected by IFN-β treatment, the expression of CD86 and CD83 induced upon A. fumigatus challenge was enhanced in IFN-β-conditioned DCs, which also showed an increased expression of PAK5 IL-27 and IL-12p70, members of IL-12 family. Through these modifications, IFN-β improved the capacity of DCs to promote an anti-Aspergillus T helper type 1 response, as evaluated by mixed leucocyte reaction, which plays a crucial role in the control of invasive aspergillosis. Our results identified a novel effect of IFN-β on anti-Aspergillus immune responses which, in turn, might open new perspectives on the use of IFN-β in immunotherapy for fungal infections aimed at enhancing the immunological functions of DCs. Aspergillus fumigatus ( A. fumigatus) conidia are ubiquitous in the environment.

As COX-2 expression crucially depends on p50 homodimer binding to distinct promotor sites,[19-21] this pathway might also be responsible for up-regulation of COX-2 expression under the conditions used in the present study. Further investigations will have to elucidate the exact molecular mechanisms leading to this potential converse effect of n-butyrate on different NF-κB signalling pathways. In conclusion, we have demonstrated Y-27632 in vitro that n-butyrate potently up-regulates expression of key enzymes and receptors of the eicosanoid pathway when activated via bacterial stimulation, leading to an increased release of PGE2, 15d-PGJ2,

LTB4 and thromboxane B2. Through selective induction of several eicosanoid mediators and up-regulation of its receptors we speculate that such effects of SCFAs might contribute to the generation of the gut intrinsic milieu, thereby specifically regulating the local gastrointestinal

immune response. Figure S2. n-Butyrate up-regulates cyclo-oxygenase 2 (COX-2) expression in monocytes after both this website lipopolysaccharide (LPS) and Staphylococcus aureus cell (SAC) stimulation as demonstrated by Western blot. Results are representative of four independent experiments. Table S1. Names of investigated genes. “
“Type 1 diabetes results from a T cell-mediated destruction of insulin-producing pancreatic β cells. Little is known on local factors contributing to migration of T cells to pancreatic tissue. We recently demonstrated evidence of viral infection in β cells in several recent-onset type 1 diabetes patients. Islet inflammation was analysed in a series of new- or recent-onset type 1 diabetic patients and non-diabetic control subjects. Autoimmune T cell reactivity was studied in lymphocytes derived from pancreas-draining lymph nodes of one recent-onset type 1 diabetes patient in partial clinical remission. Insulitic lesions were characterized 4-Aminobutyrate aminotransferase by presence of β cells, elevated levels

of the chemokine CXCL10 and infiltration of lymphocytes expressing the corresponding chemokine receptor CXCR3 in all pancreatic lesions of type 1 diabetes patients, regardless of enterovirus infection of β cells. CXCR3 and CXCL10 were undetectable in pancreata of non-diabetic control subjects. T cells isolated from draining lymph nodes of a recent-onset patient with virally infected β cells and in clinical remission reacted with multiple islet autoantigens and displayed a mixed interferon (IFN)-γ/interleukin (IL)-10 cytokine pattern. Our data point to CXCL10 as an important cytokine in distressed islets that may contribute to inflammation leading to insulitis and β cell destruction, regardless of local viral infection. We demonstrate further pro- and anti-inflammatory islet autoreactivity, indicating that different adaptive and innate immune responses may contribute to insulitis and β cell destruction.

Indeed, as shown in Fig. 6B similar Foxp3 staining could be observed

in the lamina propria of TNBS-treated mice that received PI when compared to TNBS-treated mice that received saline suggesting that Foxp3+ T cells are normally present in the lamina propria of PI-treated mice. Our data demonstrate that the phospholipid PI inhibits T-cell proliferation and Proteases inhibitor differentiation but does not affect Treg differentiation. In vitro experiments established that PI strongly inhibited PMA-stimulated T-cell activation. Moreover, T cells stimulated with plate-bound anti-CD3 and soluble anti-CD28 antibodies were also effectively inhibited by PI. The latter experiments excluded that PI acted through interference with processes such as transmembrane diffusion of PMA. Inhibition was dose dependent and did not involve anergy, apoptosis or deletion as reflected by the fact that suppression learn more was reversible when PI was removed (Supporting Information Fig. 3). Moreover, suppression was not restricted to a specific subset of T cells as both CD4+ and CD8+ T-cell activation was inhibited. In contrast, PI did not affect T-cell proliferation indirectly by inhibiting antigen-presenting DCs, as loading of DCs in the presence of PI before co-incubation did not

inhibit T-cell activation. These data strongly implicate an effect of PI on the intracellular pathways that are associated with T-cell activation. By determining phosphorylation activity of various intracellular checkpoints of T-cell activation we established that PI exerted its effects on the PKC–MAPK pathway. As we found decreased ERK1/2, P38 and JNK phosphorylation upon PI treatment, we hypothesize that PI targets ADP ribosylation factor one of the common inositol lipid pathways that precedes these downstream routes. Recently, it was established that inositol lipid signaling is regulated through cellular expression of a class of PI-transfer proteins PI-TPα and PI-TPβ 11. Modulation of this signaling has been associated with changes in cellular proliferation. In particular, overexpression of PI-TPα in fibroblasts induces an increased growth rate, whereas the growth rate of PI-TPβ overexpressing cells

is decreased 12, 13. Future studies into the specific regulation of expression of such transfer proteins may lead to the development of novel PI-based immunosuppressants. The inhibition of signal transduction in T cells cultured with PI led to reduced IL-2 mRNA expression and low levels of IL-2 protein release, which abrogated T-cell proliferation. In consequence, PI inhibited inflammatory CD4+ Th cell proliferation and cytokine release. Crucially, Foxp3+ Treg differentiation was not aborted by addition of PI, which is a pivotal characteristic for a potential immunosuppressant. As such, the immunosuppressive effects of PI share more similarity with the inhibitor rapamycin, which preserves Foxp3+ T cells while inhibiting inflammatory responses than with cyclosporine, which suppresses both inflammatory and Tregs 14, 15.

Briefly, for the last 18 h of culture, 20 μl 3H-thymidine (NEN PLX3397 ic50 Life Science Products, Amsterdam, The Netherlands) at a concentration of 5μCI/ml was added. 3H-thymidine incorporation was determined by liquid scintillation counting, expressed as counts per minute (CPM) according to standard procedures. For data storage and management, Microsoft Excel (Microsoft, Redmond, WA, USA) was used. Graphic presentation was performed with GraphPad Prism version 5.00 (GraphPad Software, San Diego, CA, USA), and statistical analysis was performed

with SPSS version 15.0 (IBM, SPSS, Armonk, NY, USA). Data are shown as median with range unless stated otherwise. Data were analysed by Wilcoxon signed ranks test. Statistical significance was denoted at P < 0.05. We first investigated the expression of the four PARs at mRNA levels on freshly isolated naïve monocytes. Primers specific for PAR-1, PAR-2 and PAR-3 yielded bands of PD0325901 mw the expected respective size (Fig. 1). Only a faint band of PAR-4 amplification product was observed. Analysis of monocyte RNA without reverse transcriptase did not lead to amplification of any product, indicating that the PCR products obtained

were not due to genomic DNA contamination (data not shown). In all cases, positive control expression of β-actin at mRNA level was found. We next investigated expression of the four PARs and TF at the protein level on freshly isolated naïve CD14+ monocytes. As an example, freshly isolated naïve CD14+ monocytes showed clear expression of PAR-1, PAR-3 and PAR-4, but not of PAR-2 and TF (Fig. 2). The expression profile is representative for the other individual donors. These results support that PAR-1, PAR-3 and PAR-4 mediated cell signalling in naïve monocytes are possible. To test whether PAR- and TF expression on naïve CD14+ monocytes changed upon stimulation with possible PAR signalling molecules changed, PAR and TF expressions were evaluated in naïve CD14+ monocytes

cultured for 24 h in the presence of FVIIa, the binary TF-FVIIa complex, the binary TF-FVIIa complex with FX, FX, FXa, thrombin and as a positive control LPS. As shown in Figs. 3 and 4, both the percentage positive PAR-1, PAR-3 and PAR-4 expressing naïve monocytes and the mean fluorescence for PAR-1, PAR-3, and Olopatadine PAR-4 were not altered. Percentage positive monocytes for medium conditions were 97% (range 4), 5.84% (range 1.1), and 99.9% (range 0.1), and 3.2% (range 2.86) for PAR-1, PAR-3 and PAR-4, respectively. The median mean fluorescence for medium conditions was 73.5 (range 1), 286.5 (range 97), 183 (range 131) and 38.2 (range 13.4) for PAR-1, PAR-3 and PAR-4, respectively. Also, TF expression was evaluated on freshly isolated monocytes, and the change in expression upon the different coagulation proteases tested. TF (3.2%; range 2.86) was hardly detectable on the freshly isolated naïve monocytes (Fig. 2E).

NSG mice were either irradiated with 200 cGy or not irradiated (0 cGy) and mice from each group were then implanted with 1 mm3 fragments of human fetal thymus and liver in the renal subcapsular space. All mice were then injected intravenously with 1 × 105 to 5 × 105 CD34+ haematopoietic stem cells derived from the autologous human CD3-depleted fetal liver. Human B cell subsets were defined as follows: immature/transitional (CD10+/CD27–/CD38+/IgD–), transitional [CD10–/CD27–/CD38+/immunoglobulin (Ig)Ddim], naive (CD10–/CD27–/CD38–/IgD+) and memory (CD10–/CD27+) CD20+ B cells. The gating

strategy used to identify the human B cell subsets is shown in (a). The proportion of immature/transitional (b), transitional (c), naive Selleck R788 (d) and memory (e) CD20+ B cells is shown for the blood and spleen at 16 weeks post-implant and for human blood. *P < 0·05; **P < 0·01; ****P < 0·0001. Fig. S7. GSK-3 inhibitor Irradiation does not alter human innate immune cell development in non-obese diabetic (NOD)-scid IL2rγnull-bone marrow, liver, thymus (NSG–BLT) mice. NSG mice were irradiated with 200 cGy or not irradiated

(0 cGy) and mice from each group were then implanted with 1 mm3 fragments of human fetal thymus and liver in the renal subcapsular space. All mice were then injected intravenously with 1 × 105 to 5 × 105 CD34+ haematopoietic stem cells derived from the autologous human CD3-depleted fetal liver. Human innate immune cell subsets were defined as follows: macrophage (CD14+/CD33+), myeloid dendritic cells (mDC, CD11c+/CD33+) and plasmacytoid dendritic cells (DC) (pDC, CD123+/CD33+). The gating strategy used to identify the human innate subsets is shown in (a). The proportion of monocyte/macrophage (b), mDC (c) and pDC (d) is shown for the blood, spleen and bone marrow at 16 weeks post-implant and for human blood. **P < 0·01; ***P < 0·001. Fig. S8. Influence of the number of injected

human CD34+ haematopoietic stem cells (HSC) and T cell levels on the incidence of xeno-graft-versus-host disease (GVHD) in non-obese diabetic (NOD)-scid IL2rγnull-bone marrow, liver, thymus (NSG–BLT) mice. NSG mice were irradiated with 200 cGy and implanted with 1 mm3 fragments of human fetal thymus and liver in the renal subcapsular space and then injected Ureohydrolase intravenously with the indicated number of CD34+ HSC derived from the autologous human CD3-depleted fetal liver. (a) NSG–BLT mice were monitored for survival and the day of death compared to the number of injected HSC is shown. (b) The peripheral blood of recipient NSG mice was screened for development of human CD3+ T cells at 12 weeks after implant and compared to the day of death. (c) The incidence of GVHD was also compared for male NSG mice engrafted with either female or male donor tissues. Each point shown represents an individual mouse. Survival was monitored over 200 days after implant. Fig. S9.

For MIF stimulation, 1 × 107 spleen cells were incubated for 24 hr in RPMI-1640 medium containing 100 ng/ml recombinant MIF as described previously.29 Splenocytes (1 × 106 cells) were incubated with anti-CD74 (Santa Cruz Biotechnologies, Santa Cruz, CA), anti-CD44 (Southern Biotechnology Associates, Birmingham, AL), or anti-B220 (eBioscience, San Diego, CA) specific antibodies and analysed by flow cytometry. For Annexin-V and propidium iodide staining, cells were analysed using the Phosphatidyl Serine Detection Kit (IQ Products, Groningen, the Netherlands), according to the manufacturer’s instructions, and were analysed by FACS. Lysates extracted

from either B cells, brain hippocampi or kidneys were separated on SDS–PAGE as described previously.8 The membranes were Ribociclib manufacturer incubated with the antibodies anti-CD74, www.selleckchem.com/products/Vorinostat-saha.html anti-Bcl-2, anti-Bcl-xL (Santa Cruz Biotechnologies) and anti-β-actin (Sigma-Aldrich, Poole, UK) antibodies. Membranes were incubated with the appropriate second antibody coupled to horseradish peroxidase. Detection was performed using the enhanced chemiluminescence method. Densitometric units were determined using the NIH Image program (National Institutes of Health, Bethesda, MD). Total RNA was prepared from isolated B cells, brain hippocampi or kidneys using TRI Reagent (Molecular Research Center, Cincinnati,

OH). Complementary DNA was prepared and real-time reverse transcription-PCR was performed using the LightCycler system (Roche,

Mannheim, Germany), according to the manufacturer’s instructions. The following primer sequences were used (forward and reverse, respectively): CD74 (5′-CAACGCGACCTCATCT-3′, 5′-TGTTGCCGTACTTGGTAA-3′), CD44 (5′-GCTATCCTGGCCTACC-3′, 5′-TGTCCTACCACAACCCAACT-3′), MIF (5′-CGCTTTGTACCGTCCT-3′, 5′-CGTGCCGCT-AAAATCA-3′), Bcl-xL (5′-GGACCGCGTATCAGAG-3′, 5′-GCATTGTTCCCGTAGAG-3′), Bcl-2 (5′-CCATGTGGCTATGCG-3′, 5′-ATCAGCCACGCCTAA-3′), β-actin (5′-GTGACGTTGACATCCG-3′, 5′-CAGTAACAGTCCGCCT-3′). The levels of β-actin were used for normalizing the expression levels of Tacrolimus (FK506) the studied genes. Results are presented relative to the vehicle-treated group (considered as 100%). Statistical analysis was performed using Mann–Whitney U-test and Student’s t-test. Values of P < 0·05 were considered significant. Eight-month-old BWF1 mice with established disease were divided into three groups (n = 8 to n = 12) and injected subcutaneously with hCDR1, the scrambled peptide (both 50 μg per mouse) or vehicle alone, once a week for 10 weeks. The clinical data of three treatment experiments are summarized in Table 1. It can be seen in the table that mice treated with the vehicle or with the control peptide exhibited high levels of anti-dsDNA autoantibodies. In mice treated with hCDR1, however, these levels were significantly reduced.