Mutations in WT1 and NPHS2 genes analyzed by polymerase chain reaction (PCR) and direct sequencing. Clinical and pathological reviews were done too. Results: There

was a significant relationship between both primary creatinine and hypertension in the first visit and resistance to therapy. Pthological views of focal segmental glomerulosclerosis (FSGS), glomerular fibrosis, and glomerular sclerosis were significantly related to steroid resistance group (P < 0.001). Genetic analysis for mutations of WT1 and NPHS2 genes among 29 children with idiopathic nephrotic syndrome showed 2 and 5 different mutations in WT1 and NPHS2 genes, respectively. All of the mutations were seen Small molecule library chemical structure in steroid-resistant group. Conclusion: This study demonstrates

the importance of WT1 and NPHS2 analysis and pathological study in children with nephrotic syndrome. VACHVANICHSANONG PRAYONG, DISSANEEWATE PORNSAK, McNEIL EDWARD Prince of Songkla University Introduction: Primary vesicoureteral reflux (VUR) is usually detected when complications such as urinary tract infection (UTI), hydronephrosis, hypertension, proteinuria or chronic kidney disease (CKD) occur. However, to date, little research has been done on the association between VUR and renal Belnacasan ic50 damage and the potential impact on a child’s long-term health. Objective: To examine the association between VUR and renal damage in Thai children with VUR and determine its impact on long-term health. Materials and Methods: We retrospectively reviewed the medical records of children ≤15 years diagnosed with primary VUR at the Department of Pediatrics, Prince of Songkla University, Thailand between 1987 and 2013. Associations between age, sex, VUR grade, laterality and history of confirmed UTI with renal damage and renal complications were assessed using multiple logistic regression. Results: There were 332 patients identified during the study period; 149 boys and 183 Fossariinae girls. The median (IQR) age at the time of the DMSA scan

was 14.5 (11.0–22.9) months in boys and 30.9 (17.0–63.5) months in girls (p < 0.001). Of the 663 renal units (one patient had a single kidney) 149 had unilateral and 183 bilateral disease. The frequencies of VUR grades I, II, III, IV and V were 67, 121, 137, 140 and 50, respectively. Technetium-99 m dimercaptosuccinic acid (DMSA) renal scan abnormalities were found in 173/515 (33.6%) VUR kidneys and 6/148 (4.1%) non-VUR kidneys (p < 0.001). DMSA abnormalities were strongly associated with VUR grade (abnormal in 17.8% of VUR grades I-III vs 60.5% of VUR grades IV and V, p < 0.001). Age 1–4 years (OR:1.8, 95% CI: 1.1–2.9), age >5 years (OR:3.0, CI: 1.6–5.5) vs age <1 year, males (OR: 2.8, CI: 1.7–4.5), grade 1–3 (OR: 5.9, CI: 2.3–15.0) and grade 4–5 (OR: 41.2, CI: 16.0–105.0) vs no VUR, and multiple UTI (OR: 2.3, CI: 1.3–3.9) vs single UTI were independent risk factors for renal damage on multivariable analysis.

trachomatis infection of an immortalized primary endocervical epithelial cell (A2EN). Our data suggest that NK cells lyse C. trachomatis-infected cells more efficiently at 34 hpi, when secondary differentiation to infectious EB is at an early stage, compared with a later stage (42 hpi). The increased activity of NK cells toward early stage C. trachomatis-infected cells may be beneficial to the host by reducing the levels of infectious EBs that can be released. We also investigated the effect of NK-mediated lysis of C. trachomatis-infected cells on the level of recoverable IFUs. Curiously,

although we observed that the recoverable IFUs decreased in the presence of NK cells, the magnitude selleckchem of this decrease

was smaller than effects on cytolysis efficiency. NK cytolytic activity is primarily mediated by perforin, a pore-forming protein that acts as a channel for entry of granzymes (Reviewed in Lieberman, 2003), both of which are expressed in the NK cell line used here. Granzymes induce apoptosis see more in target cells, consistent with the membrane blebbing and cytolysis we observed when C. trachomatis-infected A2EN cells were exposed to the NK cell line (NK92MI). Therefore, while NK lysis may deprive C. trachomatis of its intracellular niche, we hypothesize that C. trachomatis may be equipped with a mechanism to survive or escape NK cell-mediated host cell lysis. Thus, we believe that our data warrants further

investigation on the these impact of NK cell activity on C. trachomatis, as this may reveal novel survival mechanisms used by this bacterium against host innate immune response. This capacity of Chlamydia is reminiscent of recent observations made with the sexually transmitted pathogen Neisseria gonorrheae, which is able to escape/suppress the effects of neutrophil-associated oxidative bursts (Johnson & Criss, 2011). Interestingly, while our data and that of Hook et al. (2004) demonstrate increased susceptibility of C. trachomatis-infected cells to NK cell lysis, Mavoungou et al. (1999) have demonstrated that NK cells purified from the peripheral blood of C. trachomatis-infected patients have reduced IFNγ release and lytic capacity. These patients included those with genital and nongenital C. trachomatis serovars. Discrepancies among existing human studies on the role of NK cells in clearing C. trachomatis may reflect heterogeneity among NK cell receptors and their host-expressed ligands. Gene polymorphism in the site encoding the human activating NK cell receptor, NKG2D, has been shown to influence NK cell activity and susceptibility to some infectious diseases (Ma et al., 2010). Polymorphisms in human MICA have also been reported and may alter susceptibility to NK cell lysis (Ahmad et al., 2002; Karacki et al., 2004; Tosh et al., 2006). In light of the recent findings by Mei et al. (2009) that C.

The lower limit appears in this case to be –40 cm and unless we allow backward jumping, that’s not very likely! Although the standard deviation remains a valid estimate of variation [1], it is less helpful for distributions that are not symmetric, and there are alternative methods for analysis that are perhaps more appropriate. Non-symmetric distributions can be presented using median and the quartile values. For example, in Figure 2, the ‘skewed’ sample can be

described as having an estimated median of 141 with an inter-quartile range of (122, 142), where 122 and 142 are the first and third quartile values (the 25 and 75 percentiles). Alternatively, we could transform the data into a form that makes it more symmetric. Values that have been calculated as a ratio, for example as ‘% control’, can Pifithrin-�� be highly skewed. This is a common method of presenting data in many experiments. In such cases, the range of possible results may be limited in the lower values (it may be impossible to obtain values that are less than 0%), but not for the larger values (easy to obtain 150%, or 300%). In such cases, the logarithm of the values may be more convenient for analysis. Rank order tests such as the Wilcoxon do not specifically test for

equality of median values, so transforming the data to a more symmetrical distribution may have an advantage. However, when presenting data in a figure, it can be helpful to present in the original scale, as a logarithmic scale is selleckchem less easy to appreciate (as can be seen in Figure 2). Although such suggestions have not received universal acceptance, and valid differences of opinion have been voiced, most guidelines advocate these procedures. An easily applied checklist for authors and editors will help their incorporation into practice. “
“Microcirculation (2010) 17, 333–347. doi: 10.1111/j.1549-8719.2010.00034.x Objective:  Chronic and acute ischemic diseases—peripheral artery disease, coronary

artery disease, stroke—result in tissue damage unless blood flow is maintained or restored in a timely manner. Mice of different strains recover from arteriolar ligation (by increasing collateral blood flow) at different speeds. We quantify the spatio-temporal patterns of microvascular 3-oxoacyl-(acyl-carrier-protein) reductase network remodeling following arteriolar ligation in different mouse strains to better understand inter-individual variability. Methods:  Whole-muscle spinotrapezius microvascular networks of mouse strains C57Bl/6, Balb/c and CD1 were imaged using confocal microscopy following ligation of feeding arterioles. Results:  Baseline arteriolar structures of C57Bl/6 and Balb/c mice feature heavily ramified arcades and unconnected dendritic trees, respectively. This network angioarchitecture identifies ischemia-protected and ischemia-vulnerable tissues; unlike C57Bl/6, downstream capillary perfusion in Balb/c spinotrapezius is lost following ligation.

Continued and extensive progress in stem cell research in both basic and pre-clinical settings should support the hope for development of NSC-based therapies for neurodegenerative diseases. This review focuses on the utility of stem cells, particularly NSCs, as substrates for structural and functional repair see more of the diseased or injured brain. Parkinson’s disease, characterized by an extensive loss of dopamine (DA) neurons in the substantia nigra pars compacta and their terminals in the striatum,

affects more than 500 000 people in the US and about 50 000 new cases are reported annually.[20, 21] While the etiology of idiopathic PD is not known, several predisposing factors for the dopamine depletion associated with the disease have been suggested, including programmed cell death, viral infection, and environmental toxins. As an effective treatment for

PD, patients have been given L-dihydroxyphenyl alanine (L-DOPA), a precursor of dopamine, but long-term administration of L-DOPA consequently produces grave side effects.[22, 23] More recently, surgical deep brain stimulation has been adopted as a successful treatment for PD patients.[24] Since the late 1980s, transplantation of human fetal ventral mesencephalic tissues into the striatum of PD patients has been used as a successful therapy for patients with advanced disease.[25-28] However, this fetal tissue transplantation has serious problems associated Poziotinib cell line with ethical and religious questions and logistics of acquiring fetal tissues. In addition, recent reports have indicated that the survival Janus kinase (JAK) of transplanted fetal mesencephalic cells in the patients’ brain was very low and it was difficult to obtain enough fetal tissues needed for transplantation.[29] To circumvent these difficulties, utilization of neurons with dopaminergic (DA) phenotype generated from ESCs, iPSCs, MSCs or NSCs could serve as a practical and effective alternative for the fetal brain tissues

for transplantation. DA neurons were generated from mouse ESCs after treatment with fibroblast growth factor 8 (FGF8) and sonic hedgehog,[30, 31] over-expression of Nurr1[32, 33] or Bcl-XL,[34] or co-culture with a mouse bone marrow stromal cell line.[35] Neurons with DA phenotype have been generated from monkey ESCs by co-culturing with mouse bone marrow stromal cells and behavioral improvement was seen in MPTP-lesioned monkeys following intra-striatal transplantation of these cells.[36] DA neurons were also generated from neural progenitor cells derived from fetal brain and induced functional recovery following brain transplantation in parkinsonian monkeys.[37] Transplantation of NSCs in the brain attenuates anatomic or functional deficits associated with injury or disease in the CNS via cell replacement, the release of specific neurotransmitters, and the production of neurotrophic factors that protect injured neurons and promote neuronal growth.

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). BVD-523 price 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 RG7204 mouse 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;

Rapamycin purchase 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.

This team will promote Asian collaborative studies concerning these important issues in the nephrology community. It is not yet clear whether creation of a common equation for estimated GFR in Asians can be achieved, but the first step of the collaboration is quite successful. Once a common eGFR equation is established

in the future, IDMS-traceable creatinine assay is important to establish the comparable eGFR values. Establishment of a mutual cooperation network among Asian countries is strongly needed to promote the project to overcome CKD, a life-threatening disease for humans. GSK2126458 mw “
“A patient with known steroid-dependent rheumatoid arthritis (RA) developed an acute symmetrical polyarthropathy of small and medium-sized joints associated with markedly elevated inflammatory markers suggestive of RA flare, on day 4

after deceased-donor renal transplantation. ABT263 The patient received standard induction immunosuppression with methylprednisolone and basiliximab, and had commenced prednisolone, tacrolimus and mycophenolate mofetil. Serological investigations and joint aspirate to exclude infective causes and crystal arthropathy were unremarkable. High-dose prednisolone (50 mg daily) resulted in partial but unsustained symptomatic improvement. On suspicion of a medication-related adverse event, tacrolimus and mycophenolate mofetil were changed to cyclosporine A and azathioprine on day 16. This was followed by rapid improvement in symptoms and normalization of inflammatory markers. Unexpected sequelae

in the early post-transplantation period create diagnostic and management challenges. Medication-related adverse events are not uncommon, and we speculate in this case on the potential for medication-induced immune system dysregulation stimulating disease activity in a Loperamide chronic autoimmune condition after introduction of new immunosuppressants. A 63-year-old male underwent deceased donor renal transplantation in May 2012. His past medical history included end stage kidney disease and haemodialysis since 2009 from post-infectious glomerulonephritis in the setting of polyarticular septic arthritis (Staphylococcus aureus) and a solitary kidney. Other relevant history included stable ischaemic heart disease, atrial fibrillation, type 2 diabetes, nephrectomy (renal cell cancer) in 1988, osteoporosis and rheumatoid arthritis (RA). The RA was diagnosed at age 28, and managed with methotrexate and prednisolone until the patient commenced haemodialysis. Methotrexate was then ceased and prednisolone continued at a minimum of 15 mg daily. Despite relatively quiescent disease he had significant joint deformity, joint destruction and bony erosions. The patient either did not tolerate or declined other disease-modifying agents such as hydroxychloroquine and had not received biologics. Deceased-donor renal transplantation was uncomplicated.

gondii infection. Therefore, this Selleck Staurosporine disparity led to an increased Tact cell elimination by the mAb in B6 mice (67%), whereas in BALB/c animals, the same treatment led to the elimination of 45.3% of Tact. Because CD25 expression is not restricted to Tregs or Tact, we analyzed CD8+, CD19+ and natural killer

(NK) cells, which are also activated during T. gondii infection and could be eliminated after depletion. As can be observed in Fig. 3, in uninfected animals from both strains, the proportion of these activated subsets was very low (<3.6%), and after depletion, a slight nonsignificant reduction was detected. At the time point of infection analyzed, the proportion of these activated populations was dramatically increased in B6, but not in BALB/c mice (Fig. 3), a pattern similar to that observed in the CD4+ subset (Fig. 1). Despite the slight increase of activated CD8+, CD19+ and NK cells in BALB/c mice after infection, treatment with PC61 before infection did not modify these proportions significantly (Fig. 3). However, see more depleted/infected B6 mice showed

a significantly reduced proportion of activated CD19+ and NK cells. Therefore, PC61 treatment before infection eliminates other activated cells, and the different pattern of depletion observed between strains is a consequence of the contrasting expansion and activation of effector cells. A summary of the effect of depletion on all cell types analyzed is shown in Table 1. Because of the potent immune response generated in B6 mice, the injection of PC61 mAb eliminates a very high proportion of most activated cell subtypes (up to 69%), but only low levels of Tregs (38.1%). Hence, it is impossible to analyze the role of Tregs in T. gondii-infected B6 animals using classical CD25 depletion experiments, and any interpretation drawn from this model, including mortality rates, could be more related to a role of activated cells than to the role of Tregs. Our results

agree with a previous report (Couper et al., 2009) triclocarban and extend the current knowledge on the effect of depletion in other cell types using an infectious model. Our results were obtained using a single low dose of mAb (200 μg); therefore, it is clear that repeated injections of the mAb or the use of higher concentrations are unnecessary and would lead to the complete elimination of all subtypes expressing CD25. Even though other activated cell subtypes are also eliminated in BALB/c mice using the same treatment, Tregs are the largest eliminated cell subtype in this strain. Thus, the results obtained by Tregs depletion with anti-CD25 mAbs could provide an insight into the role of Tregs during T. gondii infection only in the BALB/c strain. As a consequence of the contrasting immune response against the same pathogen generated by two mice strains of different haplotype, the depleted cell subtypes differ.

[18] The (−) mating type cells ultimately produce trisporic acid from methyltrisporate. On the other hand, 4-dihydrotrisporin in the (−) mating type is converted into trisporin and trisporol, both of which have to be transferred to the mating partner. In the (+) mating type cells, the trisporol is then converted into the final product, trisporic acid. The key difference between the (+) and (−) mating type partners selleck chemicals llc during trisporic acid production is the fate of 4-dihydrotrisporin: which is converted into 4-dihydromethyl

trisporate in (+) and trisporin in (−).[19] The 4-dihydrotrisporin-dehydrogenase is a key enzyme, which mediates the conversion of 4-dihydrosporin into trisporin in the (−) mating type cells. Wetzel et al. found that the activity of 4-dihydrotrisporin-dehydrogenase

ZD1839 datasheet is highly upregulated in only the (−) mating type.[20] It is interesting that the two mating types need to cooperate to complete the synthesis of trisporic acid, in which intermediate products must be interchanged. Analogy is also found in the pathway of mating hormone synthesis in the plant pathogens Phytophthora species, where the alpha2 hormone produced by the A2 mating type is transferred to the A1 mating type and serves as a precursor to produce the alpha1 hormone.[21] Convergent evolution may result in an analogous mating pheromone synthetic pathway in the two distantly related lineages.[22] Sexual reproduction is governed by a small region of the genome, called the mating type (MAT) or sex locus in fungi. The MAT locus of a single species comprises two (or more) distinct alleles or idiomorphs and in general encodes key transcription factors, including homeodomain or high-mobility group (HMG) proteins. The sex locus of the Mucorales was first identified in Phycomyces blakesleeanus.[23] Unlike MAT loci in the dikarya, from which typically include two or more genes, and in some cases multiple genes in a genomic region spanning >100 kb, the P. blakesleeanus sex locus comprises a single HMG gene. Each mating type encodes an allelic HMG gene, sexP

for the (+) and sexM for the (−) mating types respectively. Both sex genes are flanked by a putative triose phosphate transporter gene (tptA) and RNA helicase gene (rnhA), forming a unique syntenic TPT/HMG/RNA helicase gene cluster (Fig. 2). The study by Idnurm et al. found that the sexP and sexM loci segregate 1 : 1 following mating, and progeny encoding sexP only mate with isolates with sexM.[23] In addition, sexMΔ mutants of Mucor circinelloides are sterile in any combination of mating with (+) and (−) mating type strains.[24] These results further support that the single HMG gene sex locus controls sexual development in the Mucorales. A series of studies identified the sex loci in other Mucorales fungi, including M. circinelloides, M. mucedo, R. oryzae, and S. megalocarpus.

The C57BL/6 mice analyzed represent the

progeny of C57BL/6J mice bred in the UAB vivarium. The ΔD-iD DH allele mutation, which had been generated in BALB/c Selleckchem SB525334 mice [19], was backcrossed onto C57BL/6 mice for 22 generations. Both strains of mice were maintained in a specific pathogen-free barrier facility. All experiments with live mice were approved by and performed in compliance with Institutional Animal Care and Use Committee regulations. Flow cytometric analysis and cell sorting of bone marrow mononuclear cells was performed as previously described [8, 17, 19, 28]. Developing B lineage cells were identified on the basis of the surface expression of CD19, CD43, IgM, BP-1, and/or IgD (Supporting information Fig. 1). Due to the decreased expression of CD43 on early C57BL/6 B-cell progenitors when compared to BALB/c B-cell progenitors, the scheme of Melchers was used to isolate the equivalent of Hardy fractions B (B220+ cKit+, CD25−, BP-1−) and C (B220+ Raf inhibitor cKit− CD25+ and BP-1+). The following sets of monoclonal antibodies were used: For the equivalent of Hardy fractions B and C, anti-B220 (PerCP) (BD Pharmingen, San Diego, CA, USA (, anti-BP-1 (PE) (a gift from JF Kearney), and anti-IgM (Cy5) (Jackson ImmunoResearch), West Grove, PA, USA), anti-cKit (allophycocyanin) (BD Pharmingen) and anti-CD25 (FITC) (BD Pharmingen). For Hardy fractions D, E, and F, anti-CD19 (SPRD) (Southern Biotech, Birmingham, AL, USA), anti-CD43

(FITC) (BD Pharmingen), anti-IgD (PE) (Southern Biotech), and anti-IgM (Cy-5) (Jackson ImmunoResearch). Total RNA isolation, VH7183-specific VDJCμ RT-PCR amplification, cloning, sequencing, and sequence analysis was performed as previously

described [8, 17, 19]. A listing of the 577 wild-type C57BL/6 VDJCμ unique, in-frame sequences used for analysis in this work is provided in Supporting Information Table 1. A listing of 52 VDJCμ sequences from the congenic C57BL/6 IgHa ΔD-iD mature, recirculating fraction F bone MRIP marrow B-cell subset are provided in Supporting Information Table 2. Differences between populations were assessed where appropriate by Student’s t-test, two tailed; Fisher’s exact test, two tailed; χ2; or Levene’s test for the homogeneity of variance. Analysis was performed with JMP version 8 (SAS Institute, Cary, NC, USA), or with GraphPad Prism 5.03 (GraphPad Software, La Jolla, CA, USA). Means are accompanied by the SEM. The authors wish to thank Dr. Peter D. Burrows for his invaluable advice and support. This work was supported by NIH AI42732 (HWS), NIH AI48115 (HWS), NIH HD043327 (RLS), and by core facilities supported by NIH G20RR025858, P30 AR48311, P30AI027767, and P30 CA13148. The authors declare no financial or commercial conflict of interest. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset.

2A). In this experimental setting, we also observed a significant increase PI3K Inhibitor Library in vivo in the expression of the activation marker CD38 on B-cell surface after IFN-β treatment (Supporting Information Fig. 2B). Given that this protein is notoriously type I IFN inducible

[20], this result clearly shows that B lymphocytes are target of the IFN-β therapy confirming previous study by Zula et al. [21] who described a rapid activation of IFN signal transduction pathways in B cells present in unseparated blood from RRMS patients soon after IFN-β injection. In the past, we dissected the regulation of TLR7 in maturing monocyte-derived DCs and observed that its transcription was dependent on the endogenous IFN-β release [22]. Thus, to evaluate whether IFN-β therapy would modulate TLR7 expression in MS patients, we first monitored by real-time RT-PCR TLR7 level of transcription, together with that of TLR9, in MS patients versus HDs. It was of great interest to find that PBMCs obtained from MS patients display a clear defect, as compared with those of HDs, in TLR7 expression that was statistically significant (25 HDs and 45 MS patients analyzed) (Fig. 2A). This difference was not observed in the transcription

of TLR9 gene (Fig. 2B), demonstrating that in MS patients, the defective TLR7 expression is specific. Furthermore, we observed that in PBMCs isolated from the same MS patients GPCR Compound Library high throughput following 1 month of IFN-β therapy, the level of TLR7 mRNA was restored to the level observed in HDs, while that of TLR9 was not modulated (Fig. 2A and B). In the attempt to investigate which TLR7-expressing cell types in the peripheral blood might be responsible for this defect in MS patients, B cells and monocytes were purified from both HDs and MS patients at baseline and 1 month after the beginning of IFN-β therapy, since these two leukocyte populations express TLR7. Data on TLR7 expression in B cells isolated from HDs or MS (7 and 13 individuals, respectively) did not mirror the impairment observed in the context of the

mixed cell population of PBMCs (Fig. 2C and D), although a slightly enhanced level of TLR7 transcription in response to IFN-β N-acetylglucosamine-1-phosphate transferase occurred also in this experimental setting. As observed in unseparated PBMCs, TLR9 levels of B cells did not differ in HDs and MS patients irrespective of IFN-β treatment. Interestingly, when the expression of TLR7 was analyzed in monocytes of MS patients (13 individuals), a different picture appeared. Indeed, a lower TLR7 mRNA level was highlighted in monocytes from MS patients than that obtained from HD (8 individuals) and, moreover, also a robust induction was observed in response to IFN-β therapy (longitudinal analysis of 5 patients at baseline and 1 month after IFN-β treatment) (Fig. 2E). TLR9 expression was absent in monocytes (data not shown). These data for the first time indicated a defect in TLR7 signaling in monocytes of MS patients.