The CYBB gene (OMIM # 300481), which encodes gp91phox, localizes to the short arm of chromosome X at Xp21.1 [48]. Functional analysis of its transcriptional regulation has demonstrated multiple overlapping positive regulatory elements and repressors in the proximal 5′ flanking region [18, 49–51] as well as more distant 5′ regulatory elements [52]. Current literature includes 1156 unrelated kindreds with 1259 patients, and

a total of 621 different mutations, of which 368 mutations (59.3%) are unique for an individual kindred [23]. Molecular defects leading to X-linked CGD have been BTK inhibitor library identified in the coding region, introns, and (rarely) in the 5′ flanking regulatory regions of the CYBB gene [23]. Molecular changes include deletions Temsirolimus cell line (21.1%), insertions (6.6%), deletion/insertion (1.6%), nonsense (28.6%), missense (21.7%), splice site (19.7%) and regulatory region mutations (0.7%). The mutations are distributed

in a similar frequency among the exons and gene boundaries, with no preferential mechanisms or loci [23, 24, 53, 54]. Protein expression phenotypes of X-linked CGD have been classified as X91°, X91− and X91+, where the superscript denotes whether the level of gp91phox protein is undetectable, diminished or normal, respectively [55]. Among patients in whom gp91phox expression was determined by immunoblot or spectral analysis, protein levels were undetectable (X91°) in 82%, diminished (X91−) in 12% and normal (X91+) in 6% [23]. The CYBA gene (OMIM # 608508) encodes p22phox, also known as the alpha subunit or light chain of cytochrome b558 [56, 57]. Several heterogeneous mutations have been identified, and they are widely distributed throughout the gene [44]. Unlike the

heterogeneous mutations that lead to other forms of CGD, most of the autosomal defects in the NCF1 gene (OMIM # 608512) that encodes p47phox have been attributed to a single mechanism. In 35 independent patients with p47phox deficiency, the same deletion of two nucleotides was Erastin manufacturer identified in a GTGT repeat, corresponding to the first four bases of exon 2 of the NCF1 gene encoding p47phox [58, 59]. Currently, more than 300 patients described with this type of deletion [44]. This common mutation probably derives from recombination events between the NCF1 gene and an adjacent pseudogene, NCF1B, with the GT deletion [60]. A second pseudogene, NCF1C [61], shows greater homology to the functional gene, but its role in NCF1 mutation is unknown. Patients with mutations in the NCF1 gene have more benign clinical course compared to other CGD forms [22, 62]. A type II, not functional NCF1 pseudogene that presents the same sequence GTGT of the NCF1 functional gene has also been identified [63]. Mutations in the NCF2 gene that encodes p67phox (OMIM # 608515) include missense and nonsense mutations, substitutions at splice sites, a dinucleotide insertion and a variety of deletions [44, 64]. Nunoi et al.

The CD19+ CD25+ population was enriched in PB and in the inflamed synovial fluid compared with BM (Fig. 4a). Mononuclear cells in PB sorted into CD19+ CD25+ and CD19+ CD25− subsets were stimulated with EBV (3·6 × 106 copies/ culture). The CD25+ cultures responded to EBV stimulation with a significant increase in the number of immunoglobulin-producing cells, but no increase was observed in CD25– cultures of

the same RA patient (Fig. 4b). The stimulatory effect was seen on the IgM- and IgG-producing CD25+ cells. Similar EBV stimulation of the CD25+ cultures from healthy subjects had no increase of immunoglobulin-producing cells (Fig. 4c). We have previously shown that RA patients with EBV replication in BM present a better clinical response to RTX treatment.[25] Interestingly, RTX treatment was associated with a clear reduction of EBV load in patients with RA. These CB-839 mw data allowed us to speculate that active EBV might be harboured within the RTX-sensitive B-cell populations in vivo. As a consequence, in the present study we assessed the impact of EBV infection on the phenotype and function of B cells in blood and BM of patients with RA. The present study identifies the CD25+ subset of B cells to be enriched in PB of EBV+ RA patients suggesting that this CAL-101 solubility dmso population might be an important source of EBV infection for reactivation and re-infection of the RA patient.

Importantly, EBV transfection has shown an induced CD25 expression in Hodgkin’s lymphoma cells and in Burkitt’s lymphoma cells[51, 53] and in natural killer cell lines.[52] Similarly,

EBV-specific T cells can be selected using CD25.[54] In patients with RA, the CD25+ B-cell subset belongs to the memory pool of B cells, which is functionally characterized by an increased IL-10 secretion and low spontaneous immunoglobulin secretion.[43-45] We found that the CD25+ B-cell population was enriched with the cells Urocanase expressing the activation and apoptosis marker CD95. This is supported by our previous data where we observed that EBV replication gave rise to a concomitant expression of CD95 on CD19+ B cells and this might increase the sensitivity to RTX-induced depletion.[25] On the other hand, it has been shown that cells from patients with RA may be resistant to CD95-mediated apoptosis.[55] In EBV+ RA patients an increased frequency of CD25+ CD27+ memory cells are found. CD27 is shown to be critical for several steps of EBV infection, and CD27+ B cells are considered as a reservoir of EBV in the viral latency phases.[56, 57] CD27 expression has recently been identified as essential for combating EBV infection, because individuals with CD27 deficiency develop combined immunodeficiency, hypogammaglobulinaemia and persistent symptomatic EBV viraemia.[58, 59] Interestingly, it has been shown that B cells in the rheumatic synovia express latent membrane proteins 1 and 2A, the EBV-encoded proteins that provide additional survival and maturation signals to B cells.

Moreover, Selleckchem Seliciclib differences in the nature of cell stimuli has

been proposed as the reason for NETs consisting of either nuclear DNA [3], mitochondrial DNA [6] or a combination of both [16]. While the majority of reports of NET release involve the eventual rupture of the neutrophil plasma membrane [3,4,15], early S. aureus-stimulated NET release (5–60 min) has been reported to occur via a process akin to exocytosis without plasma membrane rupture [16]. Furthermore, NETs comprising only mitochondrial DNA are reported to originate from cells remaining viable [6]. Here we demonstrate, for the first time, the requirement of hypochlorous acid (HOCl) for NET release and a potential regulatory role for endogenous taurine in this process. In our studies, for NET stimulation, PMA was employed to stimulate PKC in place of the endogenous activator, diacylglycerol. PMA, which is known to stimulate NADPH oxidase generation of ROS in neutrophils, has also been reported to elicit dramatic NET release [3]. Although less physiologically relevant, PMA provides direct intracellular stimulation, removing the complication of multiple simultaneous signalling pathways and responses that are likely to be evoked in neutrophils

stimulated with more physiologically relevant receptor-mediated stimuli such as un-opsonized (Toll-like receptor; TLR) or opsonized (Fcγ-receptor) bacteria. In addition, this form of stimulation is consistent with many previously reported studies. RPMI-1640 was obtained from Biosera (Ringmer, UK), Percoll from check details GE Healthcare (Little Chalfont, UK), SYTOX green nucleic acid stain was obtained from Invitrogen (Paisley, UK), 96-well plates were from Corning (Lowell, MA, USA), InnoZyme Myeloperoxidase Activity Kit was from Calbiochem (Nottingham, UK), micrococcal nuclease was from Worthington Biochemical Corporation (Lakewood, NJ, USA) and tryptone soy agar and broth were from Oxoid (ThermoFisher Scientific, Basingstoke, UK). All other chemicals were purchased from Sigma (Gillingham, UK). Unless specified otherwise, all ex-vivo experiments were conducted using human neutrophils from medically healthy volunteers and were isolated from venous blood by discontinuous

Mephenoxalone Percoll density gradient followed by ammonium chloride lysis of red blood cells, as described previously [19]. Patients with chronic granulomatous disease (CGD) were recruited from the Department of Immunology, Birmingham Heartlands Hospital, following informed consent (West Midlands Research Ethics Committee number 10/H/1208/48). Neutrophils (1 × 105) in RPMI-1640 were seeded into bovine serum albumin (BSA)-coated [1% in phosphate-buffered saline (PBS)] 96-well plates and allowed to settle for 30 min at 37°C in the presence of the inhibitors or enzymes being tested. Cells were stimulated with 25 nM PMA or 0·75 mM HOCl and incubated for 3 h at 37°C [2]. NET-DNA was quantified using a modified version of a previously published method [20–22].

Based on the above findings, we next examined the intracellular expression of IL-10 and TGF-β1 in TLR-stimulated MLN B cells. Representative results of flow cytometry are shown in Fig. 5(a) for IL-10 and Fig. 6(a) for TGF-β1. Stimulation of TLR ligands increased the total number of B cells producing IL-10 and TGF-β1. In particular as seen from the bar diagram, CpG-DNA significantly increased the expressions of IL-10 and TGF-β1 in MLN B cells isolated

from AKR/J mice (Figs 5b, 6b), compared with those from SAMP1/Yit mice. These findings confirmed our results obtained with EIA. Previous studies have shown that CD1d and CD5 are possible cell surface markers for identification

of B cells producing IL-10 and TGF-β1,41 Selleckchem Roscovitine we therefore examined the expressions of these markers on MLN B cells stimulated by TLR ligands. Our flow cytometric results showed that B cells producing IL-10 and TGF-β1 were mainly contained in populations characterized by the cell surface markers CD1d+ from both SAMP1/Yit and AKR/J mice (Figs 5b, 6b). On the other hand, we observed the presence of the regulatory subset in both CD5+ and CD5− populations of MLN B cells. In addition, decreased expression of IL-10 and TGF-β1 in CpG-DNA-stimulated MLN B cells of SAMP1/Yit mice was confirmed by the results of real-time PCR (Figs 5c and 6c). Although the SAMP1/Yit B-cell functional Small molecule library research buy problem has been demonstrated previously,42 the plausible mechanism underlying the alteration in cell signalling pathway had not been explored. However, it was anticipated that an enlarged MLN with increased numbers of pathogenic B cells in SAMP1/Yit mice might be involved in ileitis. In our present study, we noted

an increase of CD5+/− CD1d+ IL-10+ or CD5+/− CD1d+ TGF-β1+ B-cell population in AKR/J as compared with the SAMP1/Yit mice (Figs 5a, 6a) and therefore, depending on this fact, we expect a possible ground for increased production of IL-10 and TGF-β1 produced by B cells from AKR/J mice treated with Epigenetics inhibitor TLR ligands. However, to gain detailed insight into the cell signalling events, we stimulated isolated B cells from AKR/J and SAMP1/Yit strains with CpG-DNA, as this ligand exhibited a better response than LPS for both IL-10 and TGF-β1 secretions, after which a TLR pathway focused PCR array assay was performed using total extracted RNA. Although we observed that the B cells from both strains of mice were responsive to CpG-DNA, they did not exhibit any marked difference between the B-cell types from two different strains in terms of inducing the expression of some familiar TLR pathway-related genes, e.g., Myd88, TRAF6, IRAK-1/4 (Fig. 7a).

In addition, some evidence indicates that co-activation of c-Kit 15, CD28 16, CD226 7 and CCR1 17 with FcεRI results in the modulation of the MC response. Several studies provide supporting information about the

expression of co-stimulatory cell surface molecules, including members of the B7 family (ICOSL, PD-L1 and PD-L2) 8, 10, 18 and the TNF/TNFR families (OX40L, CD153, Fas, 4-1BB and GITR) 10, 19, 20. More recently, RG-7388 in vivo considerable progress in understanding the importance of physical contact and cell surface receptors was yielded by the discovery that MCs and Tregs interact via OX40L and OX40. This axis defines a previously unrecognized mechanism controlling both MC degranulation and Treg suppression 4, 5. The finding that the interaction of OX40-expressing Tregs with OX40L-expressing MCs decreased the extent of MC degranulation in vitro and reduced the amplitude of the immediate hypersensitivity response in vivo highlights

the existence of functionally important selleck screening library MC–Treg cross-talk, raising the question of whether these cognate interactions might occur in the course of the immune response. Interestingly, the cross-talk between MCs and Tregs results in inhibition of early events induced by FcεRI triggering, such as release of histamine and proteolytic enzymes, without affecting cytokine and chemokine secretion 4. To investigate how conjugates could be established between murine and human MCs and CD4+CD25+ Tregs and how they could explain

selective T-cell-mediated modulation of MC functions, we examined the kinetics, morphological features and functional profile of cell–cell interaction and cell conjugate formation. We have reported that Tregs, but not activated T cells, can inhibit the MC allergic response without affecting cytokine release, through a cell–cell contact-dependent interaction 4. To analyze the dynamics of this process, we performed real-time imaging of MC–Treg cognate interactions. By time-lapse bright-field video microscopy, we analyzed the formation of conjugates between IgE-presensitized murine bone marrow MCs (BMMCs) and CD4+CD25+ Tregs. The time series started after Ag addition and cell behavior was observed every minute for Clomifene a total of 30 min. Each cell type was distinguished by its unique morphological characteristics. BMMCs were large (about 15–20 μm) and round, whereas Tregs were smaller (8–10 μm) with tiny cytoplasm. Under resting conditions both BMMCs and T cells were typically rounded, while during cell–cell contact both cell types became elongated and flattened (Fig. 1A). Early BMMC tethering failed to result in firm adhesion; the BMMC moved across the T cell, forming a mobile junction with a dynamic contact plane (not shown). Individual interactions showed sequential phases of adhesion, slow later movement and dynamic crawling in different proportions and duration.

The systematic review by Richter et al.48 assessed the effects of pioglitazone in the treatment of type 2 diabetes. The relevant outcomes for these guidelines are mortality (kidney disease) and morbidity (nephropathy). Overall the evidence for a positive patient-oriented outcome for the use of pioglitazone was considered not to be convincing. Three studies were identified that included endpoints relevant to the assessment of kidney disease namely, Hanefeld et al.,49 Matthews et al.50 and Schernthaner et al.51 The Hanefeld et al.49 study compared pioglitazone plus sulfonyl urea with metformin plus sulphonyl urea over 12 months in 649 people with type 2 diabetes with

a history of poorly controlled AP24534 diabetes. The pioglitazone treatment resulted in a 15% reduction in the urinary ACR compared with a 2% increase in the metformin group with both treatments giving clinically equivalent glycaemic control. The Matthews et al.50 study compared pioglitazone plus metformin with glicazide plus metformin in 630 people with poorly managed type 2 diabetes over 12 months. The pioglitazone treatment gave a 10% reduction in the ACR compared

with a 6% increase in the glicazide PD0332991 mw group with no significant difference in HbA1c. The Schernthaner et al.51 study included 1199 people with type 2 diabetes inadequately treated by diet alone (HbA between 7.5% and 11%) and aged between 35–75 years from 167 centres located across 12 European countries. Pioglitazone treatment resulted in a 19% decrease in ACR compared with 1% in the metformin group. Blood pressure was not statistically different between groups. The results

were considered to be consistent with previous studies that troglitazone but not metformin or glibenclamide reduced urinary albumin excretion rate. The systematic review by Richter et al.52 assessed the effects of rosiglitazone in the treatment of type 2 diabetes. The study by Lebovitz et al.53 was identified as including an outcome measure relevant to kidney disease. The study examined the use of rosiglitazone as a monotherapy in 493 people with type 2 diabetes over a 7 month period. Urinary albumin excretion was decreased significantly compared with the placebo. For the subgroup of people with microalbuminuria, both doses of rosiglitazone gave a reduction Tryptophan synthase in ACR from baseline of around 40%. Only a small percentage of patients were receiving antihypertensive therapy which the authors suggested indicates the effect to be a result of improved glycaemic control or a different effect of rosiglitazone. The measurement of urinary ACR was a secondary prospective outcome of the study of 203 people with type 2 diabetes by Bakris et al.54 comparing rosiglitazone with glyburide in a randomized controlled trial. RSG significantly reduced ACR from baseline and strongly correlated with changes in blood pressure and little relation to changes in FPG or HbA1c.

Proliferation was measured using MTT and BrdU kits and the role of STAT1 and chemokines was determined by use of siRNA and recombinant proteins. Stimulation of lymphatic endothelial cell cultures with IL-27 induced JAK dependent phosphorylation of STAT1 and STAT3 and inhibited lymphatic endothelial cell

proliferation and migration. Expression of CXCL10 and CXCL11, both STAT1 target genes, was profoundly up-regulated upon IL-27 stimulation, and recombinant CXCL10 and CXCL11 inhibited FGF-2-induced proliferation in vitro. siRNA targeting of STAT1 almost completely abrogated CXCL10 and CXCL11 expression as well as the proliferative effect of IL-27. IL-27 function as an anti-lymphangiogenic regulator in vitro by up-regulating chemokines and interfering with the mitogenic effect of growth factors through STAT1 activation. “
“Women with PCOS may present abnormal hemodynamic buy Seliciclib alterations and thus may develop vascular damage. This study performed LDF measurements on the skin surface around the leg to verify if beat-to-beat waveform and spectral analysis can help to discriminate the MBF characteristics between PCOS and healthy subjects. ECG and LDF signals were obtained noninvasively in PCOS (n = 16) and control (n = 8) subjects. Beat-to-beat waveform and spectral analysis was performed on the LDF signals

to obtain the AD, FDT, FRT, and REC of five frequency bands. FRT was significantly larger, AD LBH589 nmr was significantly smaller, REC of the myogenic-related band was significantly smaller and REC of the heartbeat-related band was significantly larger in the PCOS than in the control subjects. This study is the first to reveal that time-domain waveform and spectral analysis performed on skin-surface LDF signals can be used to discriminate the differences in the MBF perfusion condition and the microcirculatory regulatory activities at local vascular beds between PCOS and healthy subjects. These findings

may aid the noninvasive early detection of PCOS-induced vascular damage. “
“Please cite this paper as: Arrick and Mayhan (2010). Inhibition of Endothelin-1 Receptors Improves Impaired Nitric Oxide Synthase-Dependent Mephenoxalone Dilation of Cerebral Arterioles in Type-1 Diabetic Rats. Microcirculation17(6), 439–446. Objective:  Endothelin-1 has been implicated in the pathogenesis of many cardiovascular-related diseases, including diabetes. The goal of this study was to examine the influence of endothelin-1 receptors (ETA) in impaired responses of cerebral (pial) arterioles in type-1 diabetic rats. Methods:  We measured responses of cerebral arterioles in non-diabetic rats to endothelial nitric oxide synthase (eNOS)-dependent (ADP), neuronal nitric oxide synthase (nNOS)-dependent (N-methyl-d-aspartic acid [NMDA]) and NOS-independent (nitroglycerin) agonists before and during application of BQ-123, an ETA receptor antagonist.

Antioxidants, free radical scavengers or substances inhibiting I/R injury may reduce bladder damages caused by BOO or overdistention. As any organ in the body, the urinary bladder needs an adequate blood supply to obtain oxygen and nutrition to function normally. Ischemia with the accompanied click here hypoxia would expect to impair bladder function. Cumulated evidences have demonstrated that ischemia, hypoxia and ischemia/reperfusion (I/R), with the accompanying generation of reactive oxygen/nitrogen species, are important etiologic factors in obstructive

bladder dysfunction.1,2 The present paper reviews and summarizes the effects of ischemia and hypoxia on the energy metabolism and contractile AZD9291 function of the urinary bladder. I/R injury on the bladder and its role in chronic bladder outlet obstruction and acute overdistention are further reviewed. Chronic partial ischemia of the bladder has been shown to impair bladder function. Gill et al. have shown that bladder ischemia induced by ligation of the vesical artery impaired contractile responses of the detrusor strips.3 Lin el al. found that chronic ischemia of the bladder resulted in a decrease of bladder compliance with a reduction in the contractility of the whole bladder.4

Lit et al. further demonstrated that chronic ischemia deranged glucose metabolism of the detrusor with a reduction in glycogen content and an increase GNA12 in anaerobic metabolism, resulting in a much lower production of high-energy molecules.5 Using an atherosclerosis rabbit model, a recent study also demonstrated that chronic ischemia of the urinary bladder resulted in mitochondrial injury, fibrosis, microvasculature damage and neurodegeneration.6 Lin et al. have demonstrated that urinary bladder blood flow was reduced by outlet obstruction

and the reduction in blood flow was associated with decreased tissue level of high-energy phosphates, adenosine triphosphate (ATP) and creatine phosphate.7 They further showed that the BOO-induced blood flow reduction could be recovered gradually after relieving outlet obstruction and was in parallel with the recovery of energy producing-related mitochondrial enzyme activity and energy producing capability of the bladder.8,9 During bladder emptying, the increased intra-wall tension results in blood vessel compression, decreased blood flow and tissue hypoxia. This occurs in normal bladders; nevertheless, this phenomenon is significantly exaggerated in the obstructed hypertrophied bladder.2,10 Under conditions of increased oxidative stress, cellular and subcellular membranes become subject to attacks when the generation of free radicals outweighs the system’s ability to eliminate.

Cells were analyzed by a FACScan equipped with Cell Quest software (Becton Dickinson,

Mountain View, CA, USA). CXCL10, CCL2, and CXCL8 were measured with OptEIA™ kits (BD Pharmingen), LY294002 nmr in cell-free supernatants [sups] from resting or stimulated keratinocyte cultures, according to the manufacturer’ protocols. The plates were analyzed in an ELISA reader mod. 3550 UV Bio-Rad. Results are given as ng/106 cells ± SD. Skin biopsies were minced with a scalpel and placed in culture in complete medium plus 60 U/mL IL-2. After 2–5 days, T cells emigrated from tissue samples were collected for phenotypic and functional characterization and T-cell cloning by limiting dilution (0.6 cells per well), in the presence of irradiated allogeneic feeder cells plus 1% PHA. Subconfluent keratinocytes seeded in culture slides (BD Biosciences) were pretreated with the indicated concentrations of peptides and, then, stimulated with IFN-γ. After 24 h, cells were cocultured with 5 × 105 CFSE-stained autologous T cells clones. In blocking experiments, keratinocytes were incubated for 1 h with 10 μg/mL anti-CD54 prior to cocultures with effector T cells. After 6 h, cocultures were extensively washed in PBS, fixed in 4% paraformaldehyde, and counterstained with hematoxylin. T cells

that adhered to keratinocytes were counted in 20 casual fields for each Poziotinib in vitro condition, as fluorescent dots using a fluorescent microscope (Zeiss, Oberkochen, Germany), and average T-cell number per square millimeter ± SD was calculated. Complete RPMI with 0.5% BSA alone or supernatants (sups) from untreated or 24 h IFN-γ-stimulated keratinocyte cultures

(0.6 mL total amount) were added to the bottom chamber of 24-well Transwell chambers with uncoated 5 mm pore polycarbonate filters (Corning Janus kinase (JAK) Costar, Cambridge, MA). T autologous cells were resuspended in complete RPMI with 0.5% BSA, and 0.1 mL cell suspension (106 cells/mL) was added to the top chamber. Transwells were then incubated for 1 h at 37°C with 5% CO2. The number of cells transmigrated to the lower chamber relative to the input was measured with a FACScant by 60 s acquisition at a flow rate of 100 mL/min. The experiments were carried out in triplicate for each condition and the results are given as ng/106 cells ± SD. Five-millimeter punches of normal human skin from three healthy donors undergoing to plastic surgery. Biopsies were taken after informed consent and the study was approved by the Ethical Committee of the Istituto Dermopatico dell’Immacolata (IDI)–IRCCS (Rome, Italy). Biopsies were placed in Keratinocye Basal Medium with 0.1% normal human serum in a humidified incubator at 37°C, with enough medium to just cover the explants.

These cross-reactive T cells were found to be subdominant during the primary response, and the sequence of infection influenced the

selleck products hierarchy of these subdominant cross-reactive T cells after secondary heterologous challenge 32, 33. In our model, the immunodominant CD8+ T-cell epitope was found to be cross-reactive, but to differing degrees, following either JEV or WNV infection. Our detailed characterization of these epitope-specific responses did not demonstrate an alteration in epitope hierarchy, but rather differences in cytokine profiles and T-cell phenotype. As previous studies have elucidated a role for subdominant cross-reactive CD4+ and CD8+ T cells in protection as well as immunopathology, future experiments will address www.selleckchem.com/products/acalabrutinib.html the role of the two cross-reactive CD4+ T-cell epitopes we identified and subdominant cross-reactive CD8+ T-cell epitopes along with the immunodominant cross-reactive CD8+ T-cell epitope in secondary heterologous JEV and WNV infections 10, 11. Here, we have shown that primary infections with JEV and WNV give rise to functionally and phenotypically distinct CD8+ T-cell responses. These

differences are due to the infecting virus (JEV versus WNV) rather than the stimulating variant (WNV S9 versus JEV S9) or viral pathogenicity. The JEV/WNV cross-reactive CD4+ and CD8+ T-cell epitopes we have identified will be useful tools to study the pathogenesis of sequential heterologous flavivirus infections. Flaviviruses continue to emerge into new geographic regions of the world, giving rise

to the possibility of new patterns of sequential infection with unknown outcomes (e.g. WNV into dengue- and yellow fever virus-endemic regions of South America). Altered CD8+ T-cell effector functions between flaviviruses may to lead to immunopathology or protection upon a secondary flavivirus infection. Additional experiments are needed to determine whether cross-reactivity C1GALT1 occurs between other members of the flavivirus family and its possible impact on disease outcome. JEV strain SA14-14-2 was provided by Dr. Thomas Monath (Acambis, Inc.). JEV strain Beijing was provided by Dr. Alan Barrett (University of Texas Medical Branch, Galveston, TX, USA). WNV strain 3356 was provided by Dr. Kristen Bernard (Wadsworth Center, Albany, NY, USA). Flaviviruses were propagated and titered in Vero cells (ATCC). The EL-4 T-cell lymphoma cell line (H-2b) served as target cells. Peptide (15–19mer) arrays corresponding to the entire proteome of WNV were obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH (BEI Resources, Manassas, VA, USA). Peptide truncations (>70 or >90% purity) were obtained from AnaSpec (San Jose, CA, USA) and 21st Century Biochemicals (Marlborough, MA, USA).