Subsequently, Tan et al. also published work on competitive PCR assay quantification of WSSV in tissues, using P. monodon as the study species [10]. Li et al. reported that infection

of the crayfish Procambarus clarkii with 103–105 WSSV copies resulted in a mortality of 100% after 10 days, whereas administration of 1.06 × 106 virus copies resulted in 100% mortality after 3–7 days [3]. Recently, viral loads measured in shrimp were 109–1010 copy Paclitaxel molecular weight numbers/g of tissue at the onset of mortality [11]. White spot syndrome virus outbreaks coincide with the onset of the monsoon in Malaysia, when intense rainfall decreases the salinity of aquaculture ponds [12]. It has been suggested that acute salinity find more changes over a particular range weaken the immune systems of shrimp, making them highly vulnerable to pathogens. In shrimp culture, few studies have been performed on environmental influences over disease susceptibility and the influence of salinity on immune variables that affect disease outbreaks [13]. Fenneropenaeus indicus is one of the major commercial species. Changes in environmental factors such as salinity may regulate (both positively

and negatively) this species’ immune and biochemical variables, which could lead to greater susceptibility to, and increased mortality from, WSSV. Accordingly, we carried out the present study to investigate the role of salinity on the susceptibility of F. indicus to WSSV and the influence of WSSV on relevant metabolic and immune variables. We here report inducing acute variations in rearing salinity and their impact on biochemical and immune variables of hemolymph of F. indicus challenged with WSSV. White spot syndrome virus-free F. indicus (confirmed by WSSV PCR assay) produced Rutecarpine from specific pathogen-free brooders were collected from a shrimp farm at Tuticorin, Tamil Nadu, India and acclimatized

in the laboratory for 2 weeks before experimentation. The animals were kept in tanks with sand beds supplied with a flow-through system of sand-filtered, ozone-treated sea water at 28 ± 0.5°C. The animals were fed with commercial crumbled feed at 5% of body weight per day before and during the experiment. F. indicus in the intermolt stage were used for the study. The molt stage was identified by finding partial retraction of the epidermis on examination of the uropods [15]. The shrimp ranged from 14.7 to 20.3 g (mean ± standard deviation, 17.75 ± 3.60 g) with no significant size differences among the treatments. Before starting the experiment, the disease-free status of randomly selected samples of the experimental shrimp was screened by PCR for WSSV infection. WSSV-free F. indicus maintained at 25 g/L were selected for further studies. White spot syndrome virus inoculums were prepared from WSSV-infected shrimp (Fenneropenaeus indicus) with prominent white spots collected from shrimp farms in Tuticorin, Tamil Nadu, India.

We applied the Mann–Whitney U-test to assess the sensitivity or robustness of the results, and the results were consistent. We set the criterion for statistical significance a priori at α = 0·05. All P-values were reported to two decimal places. We have previously shown that CB CD34+ progenitor cells express functional TLR4 and respond to LPS stimulation through Eo/B CFU find more formation.[12] To confirm and extend those findings,

freshly isolated CD34+ cells were stimulated with LPS and haematopoietic cytokines for 14 days in methylcellulose cultures. Although LPS alone could not induce Eo/B CFU formation, the combination of GM-CSF (P = 0·02) and LPS resulted in a significant increase in the number of enumerable Eo/B colonies (Fig. 1a). Although the mean value was increased, IL-5-responsive Eo/B CFU formation in the presence of LPS did not reach significance (Fig 1b). We next assessed whether CD34+ cells stimulated with LPS secrete the Eo/B differentiation-inducing

cytokines, GM-CSF and IL-5, using a bioplex cytokine assay. Although none of these cytokines was found in the culture medium, CD34+ cells alone do secrete ambiently low levels of cytokines. As shown in Fig. 2(a), LPS induces significant levels of GM-CSF (P = 0·02) from CB progenitors. The mean level of IL-5 was increased in LPS-stimulated supernatant but this did not reach significance (Fig 2b). Phospho-flow cytometry is an especially valuable tool for investigating signalling HA-1077 price pathways buy AG-014699 in rare cell populations,[20] like CD34+ progenitor cells.

As it has been previously used to detect MAPK and STAT5 signalling pathways,[16] which may be involved in cytokine secretion from TLR-stimulated CB progenitor cells,[21] we investigated whether these pathways were activated by LPS stimulation of CB CD34+ cells. As shown in Fig 3, detectable levels of phosphorylated p38 MAPK were seen 5 min after LPS stimulation (P = 0·046) followed by a steady decline thereafter. Additionally, there was a trend to increased ERK 1/2 between 5 and 30 min (P = 0·06) with LPS stimulation. No significant differences in STAT5 expression, as evaluated over time, were detected in LPS-stimulated CB progenitor cells. As we show that LPS induces a significant increase in GM-CSF secretion from CB CD34+ cells (Fig 2), and that LPS can induce the rapid activation of p38 MAPK (Fig 3), we next assessed whether these pathways were involved in GM-CSF secretion by CB CD34+ cells. To do this, CD34+ cells were pre-incubated with MAPK inhibitors SB203580 (p38 MAPK inhibitor) or PD98059 (ERK 1/2 inhibitor) or a STAT5 inhibitor and GM-CSF secretion was assessed by Luminex.

By contrast, current knowledge on symbionts of nematodes is still mainly restricted to Wolbachia

and its interaction with filarial worms that lead to increased pathogenicity of the infected nematode. In this review article, we aim to highlight the main characteristics of symbionts in term of their ecology, host cell interactions, parasitism and co-evolution, in order to stimulate future research in a field that remains largely unexplored SCH772984 ic50 despite the availability of modern tools. Endosymbiosis is a symbiosis in which one symbiont dwells within the body of the other. Usually, when talking about endosymbionts, we refer to bacteria or less frequently to fungi living inside the eukaryotic cell or simply inside the body. Interestingly, the endosymbiotic

theory first articulated by the Russian botanist Konstantin Mereschkowski in 1905 (Emelyanov, 2007) describes chloroplasts, mitochondria and other organelles as originating from bacterial endosymbionts. Nearly 90 years ago, Paul Buchner, the father of symbiosis research, documented a remarkable array of both endosymbiotic fungal selleck inhibitor and bacterial associates of arthropods (Buchner, 1965). More recently, evidence has also emerged that bacterial symbionts are present in a large variety of additional eukaryotes, including nematodes, amoebae and plants (see Table 1 for a summary of selected discoveries illuminating research on symbionts). In the present review, we will focus on bacterial symbionts associated with nematodes, arthropods and free-living amoebae. Nematodes

or ‘roundworms’ form a highly successful and abundant group of organisms found in every ecosystem on Earth. Considering their ubiquity and enormous diversity, it is surprising that only relatively few examples of bacterial endosymbioses have been described in nematodes compared with amoebae and arthropods. Of these few examples, only three have been investigated in sufficient detail to unravel some Thiamet G of the biological features of the symbiotic relationship. The most extensively studied systems are the closely related Gammaproteobacteria, Photorhabdus and Xenorhabdus, which colonize the guts of Heterorhabditis and Steinemema nematodes, respectively (see Goodrich-Blair & Clarke, 2007; Herbert & Goodrich-Blair, 2007; Clarke, 2008; for in-depth reviews). The bacteria have intricate and distinct roles in the nematode’s life cycle. On entering its insect prey, the infective juvenile stage of the nematode regurgitates its bacteria into the haemolymph, which rapidly grows and kills the insect, releasing nutrients to support the growth and development of the nematode. After the adults reproduce, a process that is dependent upon symbionts, environmental cues stimulate the progeny to enter the infective juvenile stage, which becomes re-colonized with one or two bacteria through maternal inoculation.

8 million new cases of extrapulmonary tuberculosis (EPTB) were observed in 2010 worldwide (WHO, 2011). EPTB Enzalutamide clinical trial has become more common since the advent of human immunodeficiency virus (HIV) infection (Cabandugama et al., 2011; WHO, 2011). EPTB constitutes about 15–20% of TB cases and can constitute up to 50% of TB cases in HIV-infected individuals (Noussair et al., 2009; Peto

et al., 2009; Cortez et al., 2011). As India has high burden of TB cases, thus proportionately higher number of EPTB cases are also observed in this country (WHO, 2011). The diagnosis of smear-positive PTB has been considerably established, but the diagnosis of smear-negative PTB, TB–HIV co-infection and EPTB poses serious challenges (Golden & Vikram, 2005; Chang, 2007). Diagnosis of EPTB, in particular, is difficult owing to paucibacillary nature of the specimens, lack of adequate clinical sample volumes and nonuniform distribution of bacteria in those specimens as well as the disease localized in sites that are difficult to access (Chakravorty et al., 2005; Cheng et al., 2005; Galimi, 2011). Various methods are employed for the diagnosis of EPTB such as smear microscopy, culture identification, histopathology, tuberculin skin test (TST), serological assays, interferon-gamma release assays (IGRAs) and nucleic acid amplification (NAA) tests (Katoch, 2004; Lange & Mori, 2010). Smear microscopy is widely used in the diagnosis

of EPTB but has drawbacks owing to Phospholipase D1 low and variable sensitivity values (0–40%) and could not differentiate between Mycobacterium tuberculosis CCI-779 concentration and nontuberculous mycobacteria (NTM; Liu et al., 2007; Haldar et al., 2011; Derese et al., 2012). Culture identification for M. tuberculosis also has variable sensitivities (0–80%) in different extrapulmonary specimens (Padmavathy et al., 2003; Sharma & Mohan, 2004; Takahashi et al., 2008; Abbara & Davidson, 2011) with turnaround time of 4–8 weeks and requires skilful technicians (Mehta et al., 2012). Diagnosis of EPTB from tissue samples is usually made by histopathological examination that depends on the presence of granulomatous inflammation and caseous

necrosis (Liu et al., 2007; Almadi et al., 2009). However, histology does not distinguish between EPTB and infections from other granulomatous diseases such as NTM, sarcoidosis, leprosy and systemic lupus erythematosus (except for the presence of acid-fast bacilli; AFB; Bravo & Gotuzzo, 2007; Chawla et al., 2009). The TST is useful for the diagnosis of EPTB; however, false-positive reactions occur as a result of previous Bacille Calmette–Guérin (BCG) vaccination or sensitization to NTM, and false-negative results occur in the immunocompromised patients, elderly persons or overt forms of TB (Lange & Mori, 2010). The in vitro T-cell-based IGRAs have been used for the diagnosis of both latent and active TB, but these assays do not differentiate between latent and active TB infection (Pai & O’Brien, 2008).

In addition, MMPs have also been shown to be important in many malignant and inflammatory diseases with tissue destruction [7, 8]. The cleavages of non-matrix substrates including cytokines and chemokines can be decisive and direct both pro- and anti-inflammatory actions of MMPs [9]. The mechanism of action of MMPs in arterial disease and aneurysm formation has largely been attributed to their ability to proteolytically process the extracellular matrix of the aortic wall [10]. Endogenous tissue inhibitors of MMP (TIMPs) provide a balancing mechanism to prevent excessive extracellular matrix

degradation [7]. Degranulation FDA-approved Drug Library of neutrophils upon the stimuli of inflammatory and microbial virulence factors JQ1 purchase releases also oxidative proinflammatory myeloperoxidase (MPO), and a serine protease neutrophil elastase (HNE), which can further promote the cascades of inflammatory tissue destruction [11]. Series of inflammatory reactions as measured by increased serum inflammatory markers have been shown to be associated with atherosclerosis, carotid artery stenosis, and AAA [12–14]. The role of MMPs and their regulators in arterial disease remains; despite several existing publications,

unclear, and the balance between MMPs and their regulators requires further investigation. Identification of markers reflecting the MMP-system may help to identify patients with arterial disease. Thus, we investigated the serum concentrations of these markers

in the patients with degenerative arterial disease including occlusive manifestations, i.e. aorto-occlusive disease and carotid disease as well as aneurysmal manifestations, i.e. abdominal aortic aneurysms. In addition, we studied, if the values differ from those of generally healthy subjects. The study population comprised 126 patients, who underwent surgery because of symptomatic AOD (n = 18), carotid artery stenosis (n = 67) or AAA (n = 41) in the Department Palmatine of Vascular Surgery, Helsinki University Central Hospital between the years 2002–2004. Preoperative blood samples were collected from all patients before the induction of anaesthesia from an upper arm arterial line in the operation theatre. Demographic characteristics and vascular risk factors are described in Table 1. Carotid surgery was performed on symptomatic patients with a moderate (50–69%) or high-grade (70–99%) carotid stenosis. Aneurysm operations were all elective repairs for AAAs with a mean maximum diameter of 61.6 mm (range 40–112 mm). Three patients with small aneurysms had disabling claudication as well. All patients with AOD had disabling claudication caused by aortoiliac lesions, which were so extended that endovascular treatment was not feasible. None of the patients had chronic critical limb ischaemia. The serum reference values were determined from samples provided by healthy blood donors (n = 100) collected by the Finnish Red Cross, Oulu, Finland.

She also developed new-onset diabetes after PD0325901 molecular weight transplantation (NODAT) that resolved after her corticosteroid dose was lowered. Although asymptomatic bacteriuria was documented on at least one occasion over the next year, she remained well until July 2011, when she presented with dysuria, fevers and pain over the allograft. A fully susceptible Escherichia coli was isolated from blood and urine cultures and a small collection at the antero-inferior pole of the allograft was noted on ultrasonography. She was treated with intravenous ceftriaxone and

discharged with a 21-day course of oral cephalexin. She was admitted again 18 days later with similar symptoms. Multiple blood cultures did not reveal a pathogen and Pseudomonas aeruginosa was isolated from urine. The lymphocoele was aspirated and the fluid had a few mononuclear cells seen on microscopy, but was sterile on culture. She was treated initially with ceftriaxone and changed to a 2-week course of oral ciprofloxacin after the P. aeruginosa was identified. Further episodes of pyrexia in August 2011 were investigated extensively with negative blood and urine cultures. Quantiferon testing showed low mitogen response with no evidence of active tuberculosis,

reflecting subnormal immune response. Mycobacterial urine cultures were also negative. A gallium scan revealed mild asymmetrical activity in left iliac fossa that localized to the perinephric lymphocoele. Each febrile episode responded to antibiotics Romidepsin purchase directed against common urinary pathogens. E. coli, P. aeruginosa and mixed coliforms were detected on five occasions over the next 6 months. During

this period she Immune system received a 2-week course of parenteral piperacillin/tazobactam as an outpatient, followed by a 4-week course of ciprofloxacin. The latter isolates were now resistant to ciprofloxacin. From March 2012 onwards, the only organism isolated was Klebsiella pneumoniae that was resistant to all commonly available oral antibiotics. She received a further 4-week course of intravenous piperacillin/tazobactam. The diagnosis of malakoplakia was made in May 2012, following biopsy of a allograft parenchymal lesion seen on computed tomography (CT) that consisted of oedematous, multi-focal, septated high-attenuation foci measuring 5.4 × 3.4 cm and 4.7 × 4.6 cm. The bladder lining was also noted to be abnormal, with a trabeculated appearance with high attenuation signal. An urgent biopsy of the lesion excluded active malignancy and confirmed parenchymal malakoplakia with CD68+ histiocytic response and intracytoplasmic basophilic microcalcifications, with typical targetoid/owl’s eye appearances characteristic of Michaelis–Gutmann (MG) bodies (see Fig. 1).

Growth curves were generated as described in Vohra & Poxton (2011), and culture supernatants were collected by centrifugation at 13 000 g for 1 min. Supernatants were collected at 8 and 12 h (late exponential phase) and 20 and 24 h (stationary phase). The SLP, flagella and HSP preparations Cabozantinib mouse were visualized on SDS-PAGE gels stained with colloidal Coomassie blue stain G250 (Severn Biotech), and Western blots were performed with rabbit antiserum

prepared against whole UV-killed cells of C. difficile (McCoubrey & Poxton, 2001). The protein concentrations in the preparations were determined using the Bradford reagent (Sigma-Aldrich). The quantities of toxin A and toxin B were determined as described in Vohra & Poxton (2011). Endotoxin contamination in the antigen preparations was determined by an end-point LAL

assay using the Pyrochrome® reagent (Associates of Cape Cod) as per the manufacturer’s instructions. THP-1 cells (European Collection Of Animal Cell MK-2206 order Cultures, ECACC 88081201) were cultured in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% heat-inactivated foetal bovine serum, 6 mM l-glutamine, 10 mM HEPES with 100 U mL−1 penicillin and 10 μg mL−1 streptomycin (sRPMI) at 37 °C in 5% CO2. Monocytic THP-1 cells at a density of 5 × 105 cells mL−1 were incubated with PMA (Sigma-Aldrich) at 10 and 50 ng mL−1 at 37 °C for 24 h for differentiation into macrophage-like adherent cells. Immunofluorescence analysis was performed on the BD FACSCalibur (BD Biosciences) machine, and differentiation

was confirmed using FITC anti-human CD4 antibody and APC anti-human CD11b antibody (eBioscience) and also visually under a microscope. The data were analysed using the Flowjo 9.0 software. Macrophage-like cells were washed with several washes of prewarmed PBS and subsequently challenged with 100 μL of the C. difficile antigens prepared in sRPMI at concentrations of 5 and 10 μg mL−1. For the challenge with culture supernatants, 100 μL supernatant was added to the macrophage-like cells for 3 h, following which the cells were washed and the culture supernatants were replaced with fresh sRPMI. LPS from E. coli R1 (100 ng mL−1) was used as a control. The optimum times for detection of the different cytokines were determined by repeated collection of supernatants at 4 and 24 h (results ID-8 not shown), and these were found to be 4 h for TNF-α and 24 h for IL-1β, IL-6, IL-8, IL-10 and IL-12p70. The supernatants were stored at −20 °C until use. In-house ELISAs were developed and standardized for the quantification of TNF-α, IL-1β, IL-6, IL-8, IL-10 and IL-12p70. The details of the antibodies and the amounts used are described in Table 1. From repeated assays, the ELISAs were found to be suitable to detect cytokines in the range of 32 ng mL−1–31.25 pg mL−1. Recombinant proteins used as standards for TNF-α, IL-1β, IL-6, IL-10 and IL-12p70 were obtained from PeproTech and that for IL-8 was obtained from eBiosciences.

It was noted that the punctate immunostaining for MSA-1 was accompanied by sparse CD13 staining and always in juxtaposition to redistributed iDCs. We have previously shown that maturation of splenic iDC from naïve calves in vitro results in the loss of CD13 expression and gain in capacity to present antigen (12,41). Thus, similar to the P. chabaudi model in mice (23), these results

support the hypothesis that iDC mature during processing of the parasite and migrate as antigen-presenting cells to lymphocyte-rich domains. The spleen-dependent innate response of naïve find protocol calves to infection with B. bovis is also characterized by early IL-12 production with subsequent IL-10 modulation (6), the major sources of which in cattle are iDCs and monocytes/macrophages, respectively (8,14,42). We have also shown that monocytes/macrophages of cattle can produce NO with direct babesiacidal activity (14,27,43). It was interesting to note that following haemoparasitic infection, intense acute hyperplasia of monocytes/macrophages is restricted to the red pulp of both mice (23) and calves (present study). Thus, in addition to regulatory function through cytokine production, our collective findings are consistent with monocytes/macrophages acting as effector cells in close juxtaposition with infected erythrocytes as they enter

the splenic sinuses. Regarding the distribution of small leucocytes, dual-labelling experiments demonstrated acute progressive accumulation of numerous CD3+ CD4− cells and TcR1+ WC1− cells within the red Smoothened Agonist datasheet pulp. Thus, it is likely that at least a portion of these accumulated Methane monooxygenase lymphocytes were WC1−γδ T cells. The role of these cells is still not clear but as bovine WC1−γδ T cells express CD2 and CD8, can produce

IFN-γ in response to cytokine stimulation, and are found in largest proportion in the spleen and intestine (15,16,44,45), it is intriguing to consider the possibility that cells with this phenotype might be the bovine functional equivalent of NKT cells (46–48). If so, then the observed accumulation of these cells in the red pulp of naïve calves infected with B. bovis is consistent with their expected role in the transition from innate to acquired immunity. Our results are in agreement with previous reports (49,50) that demonstrate relatively small accumulations of WC1+γδ T cells within the splenic marginal zones of uninfected calves. The splenic decrease in WC1+γδ T cells during the acute response of calves to B. bovis infection may indicate their activation within the marginal zone is followed by redistribution to effector sites outside of the spleen. Indeed, several reports indicate WC1+γδ T cells are most numerous and reactive within the blood of young calves (45,49,51–53).

Finally, by using primary microglia from IL-12 receptor β1-deficient (IL-12Rβ1−/−)

and IL-12Rβ2−/− mice, we demonstrate that IL-12 induces the expression of IL-7 in microglia and macrophages via both IL-12Rβ2 and IL-12Rβ1. These studies delineate a novel biological function of IL-12 that is absent in IL-23 and other p40 family members. “
“Similarly to Helicobacter BGB324 order pylori but unlike Vibrio cholerae O1/O139, Campylobacter jejuni is non-motile at 20°C but highly motile at ≥37°C. The bacterium C. jejuni has one of the highest swimming speeds reported (>100 μm/s), especially at 42°C. Straight and spiral bacterial shapes share the same motility. C. jejuni has a unique structure in the flagellate polar region, which is characterized by a cup-like structure (beneath the inner membrane), a funnel shape (opening onto the polar surface) and less dense space (cytoplasm). Other Campylobacter species (coli, fetus, and lari) have similar motility and flagellate polar structures, albeit with slight differences. This is especially true for Campylobacter fetus, which has a flagellum only at one pole and a cup-like structure composed of two membranes. With the recently increasing consumption of poultry https://www.selleckchem.com/products/pexidartinib-plx3397.html and poultry products [1-3], Campylobacter, mainly C. jejuni, are the leading cause of bacterial food poisoning in Japan and in many other countries. In Japan, eating of raw animal products such

as chicken meat (“sasami”), chicken liver and cow liver is associated with Campylobacter infections. This organism is also one of the important causes of travelers’ diarrhea [4]. C. jejuni infection commonly causes enteritis, which can manifest as watery diarrhea or bloody Pyruvate dehydrogenase diarrhea with fever and abdominal cramps [5, 6]. It is also associated with systemic infections such as bacteremia and GBS [6, 7]. Death is rare [5]. In contrast to humans, C. jejuni are part of the normal flora of the intestines of chickens (which have a higher

body temperature, 42°C, than do humans) and are secreted into their stools. This organism almost never causes intestinal diseases in chickens [8]. C. coli is also associated with human infection, accounting for 1–25% of them [3]. Campylobacter jejuni is spiral in shape, has a single flagellum at each pole and exhibits high motility, this last feature being required for its colonization of animal and human test subjects [9]; motility is also important for C. jejuni adherence and invasion in vitro [10]. Over 40 genes are involved in biogenesis and assembly of C. jejuni flagella [11]; however, the bacterial polar structures responsible for their extremely high motility are not known. In this study, we examined the structures in the flagellate polar region of C. jejuni (and other Campylobacter species) by scanning and transmission electron microscopy to gain a better understanding of C. jejuni motility.

2c and d). However, in response to

the peptide pools of RD15 and its individual this website ORFs, PBMC of TB patients showed weak responses in IFN-γ assays (<40% positive responders) (Fig. 2c), whereas PBMC from healthy subjects showed strong responses to the peptide pool of RD15 (positive responders=83%), moderate responses to RD1501, RD1502, RD1504–RD1506 and RD1511–RD1515 (positive responders=42–56%) (Fig. 2d) and weak responses to the remaining ORFs (<40% positive responders). The statistical analysis of the results showed that positive responses induced by RD15, RD1502, RD1504, RD1505 and RD1511–RD1515 were significantly higher (P<0.05) in healthy subjects than in TB patients (Fig. 2c and d). With respect to IL-10 secretion in response to complex mycobacterial antigens, moderate responses were observed

with MT-CF and strong responses with M. bovis BCG in both TB patients (positive responders=50% and 90%, respectively) and healthy subjects (positive responders =50% and 90%, respectively) (Fig. 3a and b). However, in response to all peptide pools, IL-10 secretion by PBMC in TB patients and healthy subjects was weak (<40% positive responders), except for a moderate response to RD1508 and RD15 in TB patients and healthy subjects, respectively (positive responders=40% and 42%, respectively) (Fig. 3c and d). The analyses of IFN-γ : IL-10 ratios revealed that the complex mycobacterial antigens MT-CF and M. bovis BCG induced strong Th1 biases, which were stronger in both TB patients and healthy Resminostat subjects in response selleck chemical to MT-CF (median IFN-γ : IL-10 ratios=162 and 225, respectively) than M.

bovis BCG (median IFN-γ : IL-10 ratios=59 and 61, respectively) (Fig. 4a and b). The peptide pool of RD1 also induced strong Th1 biases in both TB patients and healthy subjects (median IFN-γ : IL-10 ratios=57 and 34, respectively) (Fig. 4c and d). However, peptide pools of RD15 and its individual ORFs exhibited neither Th1 nor anti-inflammatory biases in TB patients (median IFN-γ : IL-10 ratios=0.8–1.0), except for a weak Th1 bias to RD1504 (median IFN-γ : IL-10 ratios=2.0) (Fig. 4c), whereas all of these peptide pools, except RD1507 (median IFN-γ : IL-10 ratios=1.0), showed Th1 biases in healthy subjects (IFN-γ : IL-10 ratios=3–54) (Fig. 4d). In particular, strong Th1 biases were observed with RD15 and RD1504 (IFN-γ : IL-10 ratios=54 and 40, respectively) (Fig. 4d), and moderate Th1 biases with RD1502, RD1505, RD1506 and RD1511–RD1514 (IFN-γ : IL-10 ratios=10–16) (Fig. 4d). Furthermore, the IFN-γ : IL-10 ratios induced by all the peptide pools, except for RD1, RD1501, RD1507 and RD1509, were significantly higher in healthy subjects than in TB patients (P<0.05) (Fig. 4c and d). In this study, cellular immune responses to the ORFs of RD15 were analyzed with PBMC obtained from pulmonary TB patients and M.