3A), and a dramatic reduction of blood flow (Fig. 3B). Brain edema/swelling was documented in infected WT mice during acute ECM by measuring three distances (Fig. 3A), namely line 1 from the pituitary gland to Sylvius aqueduct, line 2 crossing the medial cerebellar nucleus and line 3 stemming from the cerebellar obex [30]. PbA-infected WT mice showed increased distance 1, indicative of brain stem swelling, and cerebellum compression documented by distance 2 reduction and distance 3 increase, as compared with noninfected mice (Fig. 3D–F), in agreement with the data from Penet et al. [30]. We document

buy AUY-922 here, for the first time, that IFN-γR1−/− mice present unaltered MRI/MRA signals upon PbA infection, with no change in cerebral vasculature nor significant alteration of the metric parameters, as compared with noninfected WT mice (Fig. 3B–F), in line with their ECM-resistant phenotype. IFNAR1−/− mice presented a intermediate phenotype, with hyper-intense signal corresponding to some swelling at the corpus callosum, modest alterations of cerebellar structure, and lower brain stem swelling that were not significantly different from PbA-infected WT mice, while the blood

flow reduction was more heterogeneous, affecting only limited areas of the brain in these mice (Fig. 3B–F). Therefore, IFN-γR1−/− mice present no MRI/MRA detectable brain alteration, confirming that type II IFN-γ signaling is critically involved in microvascular obstruction development and Midostaurin supplier ischemic brain damage consecutive to PbA infection, while the contribution of the type I IFN-α/β pathway is of lesser importance. In order

to validate the functional data obtained by MRI/MRA, we further investigated the brain microvascular lesions on day 7 after blood-stage PbA infection. Microscopically, the brain vascular blood flow perturbation in PbA-infected WT mice was associated to microvascular lesions, with perivascular hemorrhage and intravascular accumulation of mononuclear cells and erythrocytes (Fig. 4A). many These parameters were reduced in PbA-infected IFNAR1−/− mice and absent in IFN-γR1−/− mice (Fig. 4A). The brain microvascular obstruction severity and local hemorrhage was assessed semiquantitatively and a significant reduction of brain pathology was documented in IFNAR1−/− mice, with an absence of pathology in IFN-γR1−/− mice (Fig. 4B). Thus, brain microscopic examination was in agreement with MRI results. Similarly, the perivascular hemorrhage and mononuclear cells and erythrocytes sequestration seen in WT mice after PbA sporozoite-initiated infection were reduced in IFNAR1−/− mice and furthermore in IFN-γR1−/− mice (data not shown). In mice, as in human, severe malaria can be associated with respiratory distress characterized by inflammatory-mediated increased capillary permeability or endothelial damage [34-37].

Using TEM, the number of neutrophils and MCs were counted on two intestinal grids for each infected fish. The number of each type of granulocyte was determined in an area measuring 1800 μm2 in close proximity to the point of cestode attachment (i.e. the interface region) and in a second area measuring 1800 μm2 at a distance of approximately 200 μm from the site of cestode attachment. Prior to analysis, the Gaussian distributions (i.e. normality) Nivolumab purchase and the homogeneity of variances of the data were assessed; the data were subsequently square

root transformed to meet these assumptions. Using the software package Statistica 7, anovas (Statistica 7, Praha, Cech Republic) were performed to detect significant differences in the number of granulocytes determined from the uninfected and infected tench and in the abundance of neutrophils and MCs at the point of cestode attachment and then at a distance of 200 μm away. Bonferroni post hoc tests and a P < 0·01 level of

significance were used throughout. Fourteen (60·9%) of the 23 tench were parasitized with M. wageneri; identity of the cestodes was confirmed using morphology and standard taxonomic keys. The intensity of infection ranged from 3 to 130 worms per host (39·5 ± 47·7, mean ± SD). The anterior part of the intestine bore the heaviest infections with the vast majority of tapeworms still attached with their scolices embedded within the intestinal wall (Figure 1a). Upon dissection in situ, M. wageneri were noticed in groups of variable numbers and in some portion of the host intestine the presence of more than one foci was frequent (Figure 1a). In tench gut wall, at the site Cediranib (AZD2171) of M. wageneri attachment, selleck chemical a raised plaque-like formation or round nodule encircled the firmly attached scolex (Figure 1b). Histological sections revealed that specimen of M. wageneri had penetrated by means

of bluntly truncated scolex deep into the mucosa and submucosa (Figure 2a, b) and in some instances into the muscularis layer (Figure 2c). This parasite anchoring system provided a secure attachment to the tench intestine (Figures 1a, b and 2b). At the site of attachment, the tapeworms induced necrosis, degeneration and/or loss of the epithelium (Figure 2a). M. wageneri elicited intense immune cells and fibroblasts proliferation within the thickness of the tench gut wall (Figure 2b, c). Diffuse hyperplastic inflammation was noticed in tench with few M. wageneri as well as in those harbouring numerous tapeworms (Figure 2a–c). Within the submucosa layer, beneath the point of M. wageneri scolex insertion, numerous granulocytes (e.g. neutrophils, MCs) (Figure 2d), rodlet cells (Figure 2e) and collagenous fibres were observed. Degranulation of the granulocytes, which was visible by light microscopy (Figure 2d), was common in the submucosa. Parasitized intestines were determined to have a significantly higher number of granulocytes than those that were uninfected (Table 1; anova, P < 0·01).

The perinephric haematoma seen on ultrasound underscores the risk of anticoagulation in the early post-transplant period. Evidence for treatment of APS-related renal TMA is limited to case reports and retrospective series.[8, 72] In APS-related allograft TMA (Table 4) plasma exchange has been associated with a good response in two cases,[39,

73] and may have contributed to partial renal recovery in a further two cases.[34, 38] However, a patient in the HCV/aCL transplant series died of multiorgan infarction despite plasmapheresis.[42] In the current case, TMA resolved following prompt intervention with daily plasma exchange, Navitoclax clinical trial IVIg and high dose steroids, before eventual reinstitution of warfarin. In CAPS, it is postulated that plasma exchange removes pathogenic aPL antibodies and other prothrombotic

factors.[74, 75] Plasma is generally recommended as replacement fluid,[75] although the potential for procoagulant factors in plasma to learn more exacerbate CAPS has led some to suggest albumin as the replacement fluid.[72, 76] FFP was predominantly used in this case in order to minimize the risk of bleeding from concomitant anticoagulation. In a previous case report, perioperative unfractionated heparin and plasmapheresis was associated with supratherapeutic anticoagulation and retroperitoneal haemorrhage.[77] Evidence from animal models suggests a role for complement inhibition at the C5 level in the treatment of APS.[6] Eculizumab is a monoclonal antibody blocking C5 activation approved for use in aHUS (including in transplantation[31, 32, 78]). Eculizumab has been associated with successful prevention and treatment of AbMR[28, 29] and post-transplant APS-related TMA;[33, 34, 71, 79, 80] the latter includes cases where APS-related allograft TMA was unresponsive to anticoagulation and plasma exchange, but resolved after the addition of eculizumab.[33, 71] A phase 2 clinical

trial is investigating whether eculizumab administered in the course of renal transplantation is beneficial in recipients with a pre-transplant history of CAPS (NCT01029587). Cyclin-dependent kinase 3 Finally, successful use of rituximab has been reported in conjunction with other therapies in patients with APS and renal-limited TMA,[81, 82] CAPS with renal involvement[83-85] and previous CAPS undergoing renal transplantation.[34] Renal transplantation in patients with APS may be associated with macrovascular thrombosis or TMA. Consideration should be given to the range of available therapies to address both the large vessel occlusive and microangiopathic manifestations. Based on current evidence, this includes anticoagulation in conjunction with plasma exchange (with or without use of IVIg) and/or eculizumab. Results of ongoing studies are awaited with interest. Dr Barbour is a Kidney Research UK (KRUK) Clinical Research Fellow (TF12/2011). The authors wish to thank Dr Anna Richards for some very helpful suggestions.

2 (Thy1.2)-coated microbeads (Miltenyi Biotec, Germany). T cells from Thy1.1 mice were isolated with the Pan T Cell Isolation Kit (Miltenyi Biotec). In experiments involving the transfer of Thy1.1 T cells, all donor T cells were isolated with the Pan T Cell Isolation Kit. For adoptive transfer experiments, 1–3×107 T cells were i.v. transferred into recipient mice. In brief, 5×107 cells were incubated in 1 mL of 10 μM 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE, Sigma) in PBS, 0.1% FCS for 10 min at 37°C. Labeling of cells was stopped by adding five volumes of ice-cold IMDM 10% FCS and washing three times with IMDM 10% FCS. Briefly, 2–3×107 Thy1.2-sorted splenocytes

from P14 TCRtg, P14×LMP7−/− TCRtg or P14×MECL-1−/− TCRtg mice were CFSE labeled and transferred i.v. into either naïve Thy1.1 mice or Thy1.1 mice that had been infected with 2×104 PFU LCMV-WE 24 h earlier. In total, 16 and 40 h after transfer, splenocytes were Tyrosine Kinase Inhibitor Library analyzed with a FACSCalibur™ flow cytometer after RBC-lysis with 1.66% NH4Cl w/v and staining for CD8+ cells (APC rat anti-mouse CD8a, clone 53–6.7, BD Pharmingen). To determine the percentage of transferred cells currently undergoing apoptosis versus T cells that are already dead, the splenocytes have been stained

with PerCP rat anti-mouse CD8a (clone 53–6.7, BD Pharmingen), Annexin-V-Pacific Blue (Molecular Probes) and To-Pro-3 (Molecular Probes) after RBC-lysis. In this case, acquisition was done with the LSRII™ flow cytometer (BD Biosciences). To statistically assess Smoothened Agonist differences between groups, Student’s

unpaired t-test was performed using the GraphPad software. A p-value<0.05 was considered statistically significant for all analyses. The authors thank Ulrike Beck for excellent technical assistance. John Monaco and Oliver Planz are acknowledged for contributing gene targeted and transgenic mice; Dirk Busch is acknowledged for contributing recombinant Listeria. This work was supported by grants from the German Research Foundation (DFG) No. GR1517/4-1/2 and GR1517/5-1/2. Conflict of interest: The authors declare no financial or commercial conflict of interest. Methane monooxygenase Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. “
“Interacting pathogens and hosts have evolved reciprocal adaptations whose function is to allow host exploitation (from the pathogen stand point) or minimize the cost of infection (from the host stand point). Once infected, two strategies are offered to the host: parasite clearing (resistance) and withstanding the infection while paying a low fitness cost (tolerance). In both cases, the immune system plays a central role. Interestingly, whatever the defence strategy adopted by the host, this is likely to have an effect on parasite evolution.

All experiments were conducted according to the Chinese Council on Animal Care guidelines. The heterotopic cardiac xenotransplantation model was performed by the modified cuff technique. Briefly, BIBW2992 mw a median abdominal incision was performed on the donor, and the heart graft was slowly perfused with 1.0 ml of cold heparinized saline solution (50 U/mL) through the inferior vena cava before the superior vena cava and pulmonary veins were ligated and divided. The ascending aorta and pulmonary artery were transected, and then the graft was removed from the donor. In the right side of neck of the recipient, the

external jugular vein and common carotid artery were dissected, clamped, and cut. The distal end of the external jugular vein and common carotid artery were ligated, and their proximal end were placed into the tubes (Becton Dickinson) and turned back over the cuff where tightly ligated by 8-0 nylon suture (Jinhuan, China). The incision was flushed thoroughly with heparinized saline solution (50 U/mL) in order to clean intraluminal blood clots and to prevent thrombosis after surgery. The donor heart was then transferred to the neck of the recipient, the pulmonary artery was drawn over the vein cuff, see more and a circular ligature was applied. The aorta was anastomosed to the carotid artery in a similar fashion. The beating of the grafted heart

was monitored by direct cervical palpation. The degree of pulsation was scored as follows: A, beating strongly; B, noticeable decline in the intensity of pulsation; or C, complete cessation

of cardiac impulses. Eight transplants were performed to determine heart xenograft survival time. The experimental animals were divided into three groups: group A, BALB/c mouse to BALB/c mouse isografting (syngeneic control group, Selleck Fludarabine n = 16); group B, BALB/c mouse to F344 rat xenografting (xenogeneic group, sacrificed at 24 hours post-transplantation, n = 8); and group C, BALB/c mouse to F344 rat xenografting (xenogeneic group, sacrificed at 40 hours, n = 8). In group A, eight heart graft samples were harvested at 24 hours for HE staining and quantitative real-time PCR (QRT-PCR) assay, three of which were randomly selected for microarray hybridization. Another eight heart graft samples were harvested at 40 hours for HE staining. In groups B and C, eight heart graft samples were used for HE staining and QRT-PCR assay, three of which were randomly selected for microarray hybridization. Heart graft samples were collected at each time point and fixed in 10% buffered formaldehyde, embedded in paraffin, and sectioned at 5 μm for HE staining. The ensuring morphological examination was performed using an Olympus Microscope (X51, Japan). Criteria for graft rejection included the presence of lymphocyte infiltration, hemorrhage, vasculitis, and thrombosis. Individual heart graft samples were taken randomly from each group for the microarray experiment.

Among the factors involved in iontophoretic drug transfer, the concentration and the pH of the solution, the intensity of the current applied, the duration of iontophoresis, and the nature of the skin surface (thickness, glabrous or not) play a key role [74]. Combined with laser Doppler, Ach, and SNP, iontophoresis has been widely used to assess

microvascular endothelial-dependent and -independent vasodilation, respectively [25,139]. It is of note that vasodilator iontophoresis has been proposed as a new therapy in diseases affecting skin microcirculation of the digits, like systemic this website sclerosis [102,103]. This is particularly interesting, but must be distinguished from iontophoresis as a tool to explore microvascular function, and is beyond the scope of this review. The mechanisms by which Ach iontophoresis induces vasodilation

of the microvessels remain unclear Ibrutinib in vitro [25,139]. A COX-dependent pathway seems to be predominant [41,64,105], although data are conflicting [6,29]. On the other hand, NO does not extensively contribute to the response [64,105]. Interactions between prostaglandin and NO pathways could explain the discrepancies between the results of these different studies [139]. Besides the endothelium-dependent vasodilation, iontophoresis of Ach induces C-fiber (axon reflex)-mediated vasodilation [6]. The variable effect of COX inhibition and local anesthesia [6,29] on Ach-induced vasodilation may be attributed to these different components of the response to Ach iontophoresis. One of the main issues to be taken into account with iontophoresis is the non-specific effect of the current itself, which interferes with the vasodilation potency of administered drugs. Indeed, current-induced vasodilation is observed when pure water alone is used in iontophoresis (sometimes referred Silibinin to as “galvanic response”); the reaction is more pronounced at the cathode and delayed at the anode [7,38]. The amplitude of current-induced vasodilation depends on the delivered electrical charge (i.e., the product of current intensity by

duration of application) [38] (Figure 3) and the current delivery pattern. For a similar charge, repeated applications induce more non-specific effects than continuous iontophoresis [39]. Durand et al. showed that current-induced vasodilation was abolished by local anesthesia and largely reduced after desensitization of C-nociceptive fibers by capsaicin [38], suggesting a role of neural axon reflex. Moreover, prostaglandins are likely to be essential for this axon reflex-related vasodilatation [40], mainly through the COX-1 pathway [128]. Nonetheless, the exact underlying mechanisms of the interference of current with vasodilation remain unclear. Different vehicles have been used to dilute drugs (e.g., tap water, deionized water, and saline) with various galvanic responses [139].

17 In general, duplex PCR amplification of BT2 yielded clear CP-690550 cost Scedosporium-specific bands. Although the closely related species P. desertorum was also amplified, it gave a signal exclusively with the group-specific probe PS_P on the blot. This assay was found positive in five of six

clinically relevant Scedosporium species. Non-specific signals were found for S. dehoogii strains when probes of P. apiosperma, P. boydii, and P. minutispora were applied. No other cross-reactions with non-target Scedosporium species or other clinically relevant fungi were observed. The detection limit of the PCR-RLB method was found to be 50 cells μl−1 or 0.2 pg genomic DNA. Fifty-nine sputum samples, comprising five culture-positive samples and 54 culture-negative samples, were analysed by PCR-RLB hybridisation assay (Table 1). Twenty-two of the samples proved to be negative by PCR-RLB. The PCR-RLB hybridisation assay permitted the detection of members of the P. apiosperma/P. boydii complex in 32 of 52 patients (61.5%). Pseudallescheria

apiosperma was detected in 20 samples, while P. boydii and S. aurantiacum were detected in 17 and eight samples, respectively. Only two samples were found positive for S. prolificans and P. minutispora, respectively. ICG-001 cost Eight samples contained two distinct species or three species simultaneously. Figure 1 shows a typical result of PCR-RLB for some sputum samples and for a number of Scedosporium reference strains. Four of the five Scedosporium culture-positive samples proved also to be positive with PCR-RLB hybridisation assay. All isolates of the P. boydii/P. apiosperma complex were identified morphologically,

except one strain many recovered from sample 10 which was identified as S. aurantiacum and confirmed by sequencing the ITS1-ITS2 (99% identity with NCBI sequence AJ889599 from S. aurantiacum strain IHEM 144-458) and BT2 region (100% identity with the NCBI sequence AJ888441 from S. aurantiacum strain IHEM 15-458); this last sample gave a positive signal by PCR-RLB hybridisation exclusively with the S. aurantiacum-specific probe. Considering all analysed samples, PCR-RLB yielded more positive results than culturing (47 vs. 5, respectively). Among the 54 Pseudallescheria/Scedosporium culture-negative samples analysed, 21 were also found negative by PCR-RLB. Twenty-six DNA extracts gave a positive signal with one species-specific probe, while six samples gave a positive reaction with two distinct species-specific probes and one sample with three probes. Antifungal treatment (mostly with the azoles itraconazole or voriconazole) during the months preceding the sampling took place in seven of the patients. However, for the remaining Pseudallescheria/Scedosporium culture-negative samples producing discrepant results (26 samples), the patients did not receive any antifungal treatment preceding the sampling date and Scedosporium species were never detected by culture in previous or later sputum samples.

[11] Many transcription factors [e.g. promyelocytic leukaemia zinc finger, T box transcription factor (T-bet), retinoic

acid receptor-related orphan receptor-γt and GATA-binding protein 3] that mediate the development of MHC-restricted CD4+ T-cell subsets also function in type I NKT cell subsets. The acquisition of expression of NK receptors by NKT cells during thymic maturation is driven by the transcription factor T-bet.[13] However, it selleck screening library is not yet known whether plasticity (change in function in response to an experience) is manifested among the type I NKT cell subsets. This section will focus primarily on the functional roles of the type I and type II NKT cell subsets. Activation of type I NKT cells with a strong agonist such as α-galactosylceramide (αGalCer), an exogenous marine-derived glycolipid, stimulates the rapid release of many cytokines that elicit both Th1 [interferon-γ (IFN-γ)] and Th2 [interleukin-4 (IL-4) and IL-13] responses.[6-17] The widely studied type I NKT cells are more prevalent than type Selleckchem LDE225 II NKT cells in mice than in humans,[1, 18, 19] and comprise about 50% of murine intrahepatic lymphocytes.[20-22] A major difference between the two subsets resides in their TCRs. The type I NKT cell invariant TCR is encoded predominantly by a germline Vα gene (75–88%) (Vα14/Jα18

in mice and Vα24/JαQ in humans), as well as more diverse non-germline Vβ chain genes (Vβ8.2/7/2 in mice and Vβ11 in humans).[1-19, 23-25] Type I NKT cells respond to both α- and β-linked glycolipids. The semi-invariant TCR on type I NKT cells binds to CD1d in a parallel configuration that mainly involves the α-chain.[2, 4, 15, 24] Whereas type II NKT cells comprise a minor subset in the mouse, they belong to a more predominant subset in humans.[1, Phosphoribosylglycinamide formyltransferase 26] A major

proportion of type II NKT cells recognizes a naturally occurring self antigen known as sulphatide, which is enriched in several membranes, including myelin in the central nervous system (CNS), pancreas, kidney and liver (Table 2). Generally, sulphatide-reactive type II NKT cells mediate protection from autoimmune diseases by down-regulation of inflammatory responses elicited by type I NKT cells.[27, 28] However, non-sulphatide-reactive type II NKT cells may play a pathogenic role in other diseases, such as ulcerative colitis.[29] Sulphatide-reactive type II NKT cells express oligoclonal TCRs by utilization of a limited number of Vα- and Vβ-chains. In contrast to type I NKT cells, only about 14% of TCR Vα and 13–27% of TCR Vβ chains in type II NKT cells are encoded by germline gene segments.[28] Notably, type II NKT TCRs contact their ligands primarily via their β-chain rather than the α-chain, suggesting that the TCR Vβ-chain contributes significantly to antigen fine specificity.[30] The mechanism of binding of type II NKT TCRs to antigens uses features of TCR binding shared by both type I NKT cells and conventional T cells.

TAN LI PING, MOHAN YASHINI, LIM SOO KUN, NG KOK PENG, KENG TEE CHAU, KONG WAI YEW, WONG CHEW MING, WA HAFIZ, WONG MUN HOE, LIM LI HAN, JALALONMUHALI MAISARAH University of Malaya Medical Center Introduction: Cardiovascular disease is a leading cause of death among kidney patients. Screening for cardiovascular disease is therefore thought to be an essential step in the evaluation of the kidney transplant recipient. However, controversy exists

regarding the optimal assessment technique. The American Heart Association and the American College of Cardiology advise no preoperative cardiac evaluation if the patient has a good functional status. The American Society of Nephrology on the other hand, recommends myocardial perfusion imaging as part of the evaluation. selleck kinase inhibitor MLN0128 chemical structure In Malaysia, there is currently no consensus addressing this issue. We conducted a retrospective review of cardiac assessment modalities among potential kidney transplant recipients in our hospital. Methods: All living donor kidney transplant recipients who underwent a kidney transplant

evaluation in our center from 2001 to 2013 were eligible for inclusion. Basic demographic data was collected. Key variables of interest were history of ischemic heart disease, presence of heart failure, stroke, diabetes mellitus. Information regarding methods of cardiac evaluation and results were obtained. Data was analyzed with SPSS v16.0. Results: 180 check details patients

were identified, however due to missing data only 68 patients were included in the study. 66.2% were male. Mean age was 35.8 yrs (S.D 9.69). 11.8% had diabetes mellitus and 7.4% had a history of ischemic heart disease. All patients had a screening ECG done of which 85.3% were normal while the remaining had mild abnormalities. 66 (97.1%) patients had a stress ECG which was read as normal in 86.8%. The remainder had inconclusive results. 13 patients underwent coronary angiogram of which 23% (n = 3) had significant coronary stenosis requiring PCI. All of those who required PCI had history of ischemic heart disease. Conclusion: In our single center cohort of potential kidney transplant recipients, only 0.04% required PCI for cardiac optimaization, all of whom were among patients with preexisting ischemic heart disease. Due to cost constraints, more advanced techniques for cardiac evaluation like myocardial perfusion imaging of dobutamine stress echocardiograms were not done. But in our limited sample of mostly non diabetic patients; basic cardiac evaluation including screening ECG and stress ECG appeared to be sufficient. Further follow up of post operative outcomes would be important to support this. AN GUN-HEE, YU JI HYUN, HWANG SEUN DEUK, CHUNG BYUNG HA, PARK CHEOL WHEE, YANG CHUN WOO, KIM YONG-SOO, CHOI BUM SOON Transplant Research Center, Division of Nephrology, Department of Internal Medicine, Seoul St.

At 4 weeks post-immunization, mice were sacrificed, and their spleens were removed. Splenocytes were restimulated with ESAT-6 protein in vitro, and the number of IFN-γ-secreting cells and the concentration of TNF-α in the supernatant were measured using ELISPOT and ELISA, respectively. No significant differences in the number of IFN-γ-secreting cells or the concentration of TNF-α were observed

between the two groups (Fig. 3B,C). Thus, the addition of CFP-10 to the calreticulin–ESAT-6 fusion did not provide an enhancement of the Fluorouracil ic50 ESAT-6-specific immune response. We next investigated the ability of the vaccine-induced immune response to reduce the mycobacterial burden after low-dose aerosol infection in the mouse model. Mice were C59 wnt purchase vaccinated with AdCRT–ESAT-6–CFP10 via the intranasal route and BCG via the subcutaneous route, only once as described in Materials and methods. At 4 weeks post-immunization, mice were infected with M. tuberculosis. Four weeks after challenge, the M. tuberculosis burden of infected animals was determined to evaluate the

protective efficacy in both lung and spleen. The trends were similar in both organs (Fig. 4A,B). BCG caused a reduction in CFU in both the lungs and spleen of infected animals. However, there was no significant difference between mice vaccinated with the adenovirus constructs and the saline-treated group for both organs. The high incidence of TB has Non-specific serine/threonine protein kinase stimulated interest in understanding the immune response to infection, resulting in the accelerated identification of novel immunodominant mycobacterial proteins as possible vaccine candidates. Culture filtrates and RD sequences have attracted particular interest as a source of antigens. ESAT-6, TB 10.4, CFP10, MTB12, MTB39 and Ag85 A and B have all been shown to elicit protective immune responses in various animal models of TB [12, 16, 27, 28]. Even though many strategies for vaccination increase the overall immune response, this may not be the ideal solution. When multiple antigens are presented to the immune system, they will compete for

presentation, and the antigens dominating the response will not necessarily be those most relevant for protection. Thus, a targeted approach may be ideal. It has been repeatedly demonstrated that calreticulin can enhance immune responses when linked to antigens in DNA and viral vaccines [23–26]. This suggests that the use of calreticulin may be broadly applicable as a strategy to enhance vaccine efficacy. In addition, several reports have suggested the efficacious use of vaccines against TB in mice using adenoviral vectors expressing different M. tuberculosis antigens [10]. We herein demonstrate the effects of a replication-deficient adenoviral vector that contains the M. tuberculosis ESAT-6 antigen fused to calreticulin and show that there is an increased immune response to this antigen as demonstrated by increased cytokine expression.