90% and 76% vs. 67%, respectively; Losi et al., 2007). Therefore, QFT-GIT is more sensitive

and rapid than conventional microbiological tests, and more specific than conventional TST for the diagnosis of tuberculous pleurisy. Furthermore, we evaluated M.tb-specific nested-PCR to aid the diagnosis of tuberculous pleurisy. The sensitivity and specificity were 94.8% and 90.0%, respectively, both of which were comparable to QFT-GIT. The greatest concern with molecular biology techniques is false-positives due to cross-contamination during processing. To eliminate any possibility of cross-contamination RGFP966 research buy from the positive controls, the Seeplex® MTB Nested ACE Detection kit used in this study designs the amplification sizes of the positive control PCR products differently from those of the specimen PCR products. Two patients in the non-TB group were nested-PCR positive and QFT-GIT negative, which indicated that the combined immunoassay and molecular Enzalutamide in vitro detection would improve the accuracy of diagnosis. The detailed analysis confirmed that both QFT-GIT and nested-PCR positive results increased the specificity to 100%,

with the sensitivity of up to 90.0%. In conclusion, QFT-GIT is more sensitive and rapid than conventional microbiological tests, and more specific than conventional TST in the diagnosis of tuberculous pleurisy. Thus, the combination of immunoassay and molecular detection holds promise in the clinical treatment of tuberculous pleurisy. The present study was partly supported by the National Natural Science Foundation of China (30901277), the US–China Biomedical Collaborative Research

Foundation (81161120426) and Wuxi Social Development Guiding Program (CSZ00N1229). All authors have stated that they do not have any conflict of interest. Y.G. and Q.O. contributed equally to this work. “
“T cells are exquisitely poised to respond rapidly to pathogens and have proved an instructive model for exploring the regulation of inducible genes. Individual genes respond to antigenic stimulation in different ways, and it has become Cobimetinib supplier clear that the interplay between transcription factors and the chromatin platform of individual genes governs these responses. Our understanding of the complexity of the chromatin platform and the epigenetic mechanisms that contribute to transcriptional control has expanded dramatically in recent years. These mechanisms include the presence/absence of histone modification marks, which form an epigenetic signature to mark active or inactive genes. These signatures are dynamically added or removed by epigenetic enzymes, comprising an array of histone-modifying enzymes, including the more recently recognized chromatin-associated signalling kinases. In addition, chromatin-remodelling complexes physically alter the chromatin structure to regulate chromatin accessibility to transcriptional regulatory factors.

These findings suggest that the activation of TLRs in pregnancy causes not only preterm labor and pregnancy loss, but also pre-eclampsia. The fetus is not indifferent to a viral or bacterial infection, and the immunological responses by the Selleck Decitabine maternal immune system or the placenta or fetal immune system may have important consequences on the normal development and survival of the fetus. In the following section, we will discuss some studies

related to the long-term effects of TLR ligation in the offspring. Administration of LPS to pregnant mice was shown to cause acute fetal cardiovascular depression,54 and inhibit structural development of the distal fetal mouse lung in a TLR4-dependent manner.66 Similarly, cerebral white matter damage, which is one of the biggest problems seen in preterm neonates because of its strong association with their lifetime adverse outcome, is also believed to be caused by TLR4 activation in the fetus.67 It is worthy to mention that low-dose LPS, which has no adverse effects on pregnancy outcome, dramatically increases brain injury to subsequent hypoxic–ischemic challenge in a newborn rat animal model.67 These findings ERK inhibitor are compatible with clinical findings showing that maternal exposure to bacteria not only causes preterm labor, but also

contributes to long-term adverse outcome in the offspring such as cerebral white matter damage. Adverse effects of maternal TLR3 activations were also found in fetuses in various animal models. Maternal poly(I:C) or virus exposure cause marked behavioral changes in the offspring mouse,68 which is relevant to many epidemiological studies showing that maternal exposure to virus causes not only abortion or preterm Exoribonuclease birth but also fetal schizophrenia and autism.69,70 Offspring of poly(I:C)-treated pregnant mice were shown to have less expression of brain-derived neurotrophic factor (BDNF), nerve

growth factor (NGF) and TNF-α in their placenta, liver/spleen and brain, which may represent a potential mechanism through which maternal viral infection increases a risk of such neurodevelopmental disorders.71 In contrast to the ‘adverse’ effects of maternal infection on fetus, there is a notion called ‘hygiene hypothesis’, that is, ‘adequate’ maternal microbial exposure has protective effects on neonatal allergic disease. Very recently, it was demonstrated that TLR system is also involved in this effect. Conrad et al.72 showed that an administration of non-pathogenic microbe Acinetobacter Iwoffii F78 to pregnant mice has a protective effect on postnatal asthma, and the effect was completely abolished in TLR2/3/4/7/9/null mice.

The frequency of cells producing CCL3 among Galunisertib supplier tetramer+

CD8+ T cells was also twice as high and equivalent to that of mice immunized with wt Lm (Fig. 1C) suggesting that increasing the immunizing dose of secA2−Lm restored the ability of reactivated memory CD8+ T cells to secrete CCL3 in vivo. Of note, this analysis gave comparable results on two distinct mouse genetic backgrounds and over three distinct naturally presented Lm-derived H2-Kd-restricted epitopes 19 and the H2-Kb-restricted SIINFEKL OVA-derived model epitope (Table 1). Therefore, protective immunity in mice immunized with wt and 107secA2−Lm correlates with CCL3 expression and higher numbers of effector memory CD8+ T cells. Thus, we established an original experimental system using different doses of the same mutant bacteria that do or do not prime protective immunological memory, and in which the signals integrated by the priming APCs are likely distinct. Efficient induction of long-term protective immunity requires the escape and the growth of Lm in the cytosol of infected cells 16, 20. We therefore looked for the

cell subsets that sustain active Lm growth inside their cytoplasm in vivo. To define such cells, mice were immunized i.v. with 106 or 107secA2−Lm-expressing GFP that is only expressed by viable Lm as GFP expression is rapidly lost Protease Inhibitor Library purchase upon bacterial death 16. 2.5, 5 and 10 h later, spleens were harvested and stained with cell surface markers allowing the discrimination of the different myeloid-derived cell subsets containing live bacteria (Supporting Information Figs. 2 and 3). At both early time points analyzed (2.5 and 5 h), CD8α+ cDCs were the main subset of cells expressing GFP (75.2 and 64.4 % respectively), and containing viable bacteria (Supporting Information Fig. 3A), 16), as also reported for wt Lm21. Interestingly, intracellular

staining of spleen cells using serum against Lm antigens, which detects both live and dead Lm as well as secreted bacterial antigens, showed that innate phagocytes, Ibrutinib ic50 i.e. neutrophils, inflammatory monocytes and macrophages, represented 69 and 62% of the positive spleen cells 2 and 5 h after the immunization respectively (Fig. 2 and data not shown), a result supporting their role in the uptake and the killing of Lm22. Therefore, while CD8α+ cDCs represent 20–30% of the Listeriapos cells, they are the major cell type exhibiting live Lm (65–75%), likely providing the most ‘hospitable’ intracellular environment for Lm growth in vivo. Since CD8α+ cDCs are permissive to Lm growth, it makes them likely to integrate and convey signals from cytoplasmic bacteria early after immunization. Previous reports showed that CD8α+ and CD8α− cDCs prime naïve Lm-specific CD8+ T cells with equivalent efficiency when loaded with exogenous peptide ex vivo 11.

In both cases, CD161 expression levels appeared lower in NK cells from individuals with symptomatic HCMV infection, an effect that was not perceived when groups were compared (Fig. 1). The NKR distribution pattern associated to HCMV infection in T lymphocytes resembled only partially that observed in NK cells (Fig. 2). Overall, the absolute numbers of NKR+ T cells were increased in HCMV+ children, particularly in the congenital symptomatic group. In fact, the proportions of

NKG2C+, LILRB1+, and CD161+ T cells were significantly higher in congenitally infected than in noninfected children. In addition, NKG2A+ T cells appeared also higher in children with congenital symptomatic infection, at variance with the reduced proportions of NKG2A+ NK cells in the same group. Altogether, these results point NVP-BEZ235 in vitro out that marked changes in NKR distribution, particularly an increase of NKG2C+ and LILRB1+ NK cells, are associated with congenital symptomatic HCMV infection. The putative implications of the NKG2C deletion on the response to HCMV infection are uncertain. On that basis, a genotypic analysis of NKG2C was conducted in children with symptomatic (n = 15) and asymptomatic (n = 11) congenital infection, as well

as in a control group including children with postnatal infection (n Selleck XL765 = 11) and noninfected (n = 19). The homozygous NKG2C deletion was found in a single uninfected control individual. In addition, no significant differences were found between the frequencies Resminostat of the heterozygous NKG2C+/− genotype detected in uninfected controls and children with congenital infection (42.1% versus 34.6%; p = 0.61). Altogether these results argue against a direct relation of the NKG2C deletion with the incidence of congenital HCMV infection in newborns. In line with previous reports [26, 27, 32], individual differences in NKG2C surface staining intensity were noticed (Supporting Information Fig. 1). The NKG2Cbright/intermediate expression pattern was generally

associated to HCMV infection, whereas all noninfected and ∼43% of infected children displayed a predominant NKG2Cdim phenotype. The proportions of NKG2C+ cells correlated significantly (r = 0.74; p < 0.001) with the KLR surface expression levels (MFI). The possibility that NKG2C copy number might influence the expansion of NKG2C+ cells and/or the expression levels of the receptor was addressed. To this end, the proportions and absolute numbers of NK cells bearing NKG2C, as well as its surface staining intensity, were compared after stratification for HCMV infection and the NKG2C genotype. As expected, increased proportions of NKG2C+ NK cells and higher surface levels of the KLR were detected in HCMV-positive children (Table 3); though less marked, a significant association of both parameters with the NKG2C genotype was also noticed.

A detailed phenotypic characterization of induced CD8+Foxp3+ T cells revealed high expression of classical Treg markers including CD25, GITR and CTLA4, consistent with previous reports 17, 31 and likely reflecting T-cell activation, although one study reported low CD25 expression on CD8+Foxp3+ T cells 38. Interestingly, the classical

Treg markers CD73 and CD103 were selectively expressed by induced CD8+Foxp3+ T cells, underlining that their expression is dependent on TGF-β, RA and/or Foxp3. In line with this, CD8+ T cells deficient in TGF-β signaling fail to up-regulate CD103 in a GVHD model 39, and Foxp3 has been shown to directly bind the CD103 promoter 40. However, Foxp3-independent mechanisms can also activate CD103 3, consistent with the only mildly reduced induction of CD103 expression in stimulated T cells check details from DEREG×Rag1−/−×OTI×Sf mice (Supporting Information Fig. 3C). CD8+Foxp3+ T cells only displayed little suppressive capacity compared with CD4+Foxp3+ Tregs, and CD8+Foxp3− T cells showed similarly low suppressive activity in vitro. Furthermore, adoptive transfer NVP-AUY922 nmr of induced CD8+Foxp3+ T cells did not ameliorate disease in an OVA-based allergic airway inflammation model (data not shown). Previous studies have reported the suppressive capacity of TGF-β-induced

CD8+ T cells 17, 31, 34, 38, which in principle does not contradict our data. First, several studies did not compare the strength of suppression to that of CD4+ Tregs 31, 34, 38, which depend on Foxp3 3. Second, suppressive CD8+ T cells were isolated either based on CD25 expression 17 (also broadly up-regulated on activated Foxp3− T cells, at least in the absence of IL-6), or were tested without ifoxetine further separation for suppressive function 31, 38, thereby not allowing for discrimination between Foxp3+ and Foxp3− subsets. Third, DC or agonistic αCD28

antibodies were used during in vitro differentiation in all these studies. Therefore, it cannot be formally excluded that the low suppressive function observed in our study is caused by the lack of signals provided by either DC or αCD28. However, this would underlie Foxp3-independent mechanisms, since CD8+Foxp3+ T cells can be efficiently generated without co-stimulation (Fig. 1). Strikingly, co-stimulation even represses Foxp3 induction in CD8+ T cells (Fig. 2A and B) suggesting that CD80/CD86–αCD28 would rather modulate suppressive activity in a Foxp3− subset. In sum, our results suggest that Foxp3 alone is not sufficient to confer strong suppressive activity to CD8+ T cells. Although transgenic mice with forced overexpression of Foxp3, but not WT mice, were described to harbor suppressive CD8+ T cells, Foxp3 was similarly considered as implicated but not sufficient to confer suppressive activity in a previous study 41.

, 2005). Surprisingly, S. pyogenes protein Prp does not interact with plasminogen and plasmin via lysine, however only via arginine and histidine residues (Sanderson-Smith et al., 2007). GBS bind plasminogen only by the glyceraldehyde-3-phosphate dehydrogenase (Seifert et al., 2003). Matrix metalloproteinases/metalloproteases (MMPs) are zinc- or cobalt-dependent enzymes that play a crucial role in normal function and development of CNS. This large group includes collagenases, gelatinases, stromelysins, matrilysin, membrane-type metalloproteinases, and metalloelastases. MMPs differ in cellular sources and substrate specificity, but structural domains remain the same (Kieseier et al.,

1999). MMPs may alter inflammatory cytokine activity, cleave cell surface receptors, Tamoxifen in vivo activate caspase-3, and Serine Protease inhibitor regulate other MMP family members (Kawasaki et al., 2008). Together with serine and cysteine proteases, they are able to degenerate and remodulate connective tissues. This damage leads to extravasation of blood-borne proteins, formation of brain edema, and neuronal damage. Pathogens exploit this extravasation to cross various barriers including BBB. Basal level of MMP expression in the brain is low; however, during infections, basal level of MMP expression elevates markedly. MMPs are expressed by most of the resident CNS cells such as ECs, astrocytes, microglia, and neurons together with the infiltrating immune cells (Hummel et al., 2001). Infection of

BMECs with neurotropic viruses

has been connected with decrease and/or redistribution of TJ proteins (Luabeya et al., 2000). MMP activity is highly increased in HIV-infected cells migrating into CNS. Human neuronal and glial cells infected with this virus have been shown to produce large amounts of MMP-2 (Chong et al., 1998). During the WNV infection, it has been observed that inflammatory cytokines, such as TNF-α, macrophage migration inhibitory factor, and MMP-9 play an essential Baricitinib role in BBB disruption (Wang et al., 2004; Arjona et al., 2007). It is likely that activation of MMP-9 in WNV-infected astrocytes is via MMP-3 (Verma et al., 2010). MMPs also play an important role in bacterial meningitis. In fact, MMP-8 and MMP-9, but not MMP-2 and MMP-3, are upregulated in CSF during the meningitis caused by H. influenzae, N. meningitidis, and S. pneumoniae (Leppert et al., 2000). Treponema denticola (Gaibani et al., 2010) and cell wall of Streptococcus suis strongly stimulate the production of MMP-9, whereas zinc metalloproteinase ZmpC of S. pneumoniae cleaves human MMP-9 into its active form (Oggioni et al., 2003), which leads to the BBB disruption (Jobin et al., 2006). MMP-8 is also associated with tissue destruction during Streptococcus sanguinis, N. meningitidis, and Fusobacterium nuclearum infections (Shin et al., 2008; Schubert-Unkmeir et al., 2010). Tissue destruction by N. meningitidis is a consequence of proteolysis of TJ protein occludin by MMP-8.

Magnification x40; Zeiss (AxioCam MRc5). Supplementary Figure 5. Isolation of PMNs as described in “Materials and Methods” shows a purity greater than 95%. Heparin-anticoagulated blood of 3–4 mice was pooled

and PMNs were isolated as described in “Materials and Methods”. Isolated PMNs were stained with anti-mouse Ly6G FITC (1A8) for subsequent FACS analysis. Supplementary Figure 6. The extracellular expression of CXCR2 of Lcn2-/- PMNs is significantly reduced compared to Lcn2+/+ mice. 200 μL of blood was drawn by retroorbital blood puncture of untreated Lcn2-/- and Lcn2+/+ mice at the age of 8 weeks. Whole blood was prepared for analysis of PMNs expression markers by means of FACS analysis GPCR Compound Library research buy as described in Materials and Methods. A granulocyte Ulixertinib mouse gate was set and Ly6G positive cells were analysed for CXCR2 surface expression. Data are shown as mean ± SEM of 4 mice. Student`s t-test was used for statistical analysis. “
“Characterization of the first tapeworm genome, Echinococcus multilocularis, is now nearly complete, and genome assemblies of E. granulosus, Taenia solium and Hymenolepis microstoma are in advanced draft versions. These initiatives herald the beginning of a genomic era in cestodology and

underpin a diverse set of research agendas targeting both basic and applied aspects of tapeworm biology. We discuss the progress in the genomics of these species, provide insights into the presence and composition of immunologically relevant gene families, including the antigen B- and EG95/45W families, and discuss chemogenomic approaches toward the development of novel chemotherapeutics 2-hydroxyphytanoyl-CoA lyase against cestode diseases. In addition, we discuss the evolution of tapeworm parasites and introduce the research programmes linked to genome initiatives that are aimed at understanding signalling systems involved in basic host–parasite interactions and morphogenesis. Whole-genome sequencing of cestodes

began in 2004 and currently includes the aetiological agents of alveolar echinococcosis (AE; Echinococcus multilocularis), cystic echinococcosis (CE; E. granulosus) and neurocysticercosis (NCC; Taenia solium) in addition to the rodent-hosted laboratory model, Hymenolepis microstoma. With the genomes of Echinococcus spp. near completion, and those of Taenia and Hymenolepis in advanced drafts, we have only begun to explore their full content, structure and general characteristics. Nevertheless, genomic and transcriptomic data are already advancing research in both basic and applied aspects of tapeworm biology and herald the beginning of a new era in cestodology. Here, we review the progress made in the genomics of tapeworms and provide initial insights into the presence of immunologically relevant molecules and chemogenomic approaches to the development of new vaccines.

Interestingly, we were able to show that a fusion protein can decrease the tumour burden in some, although not all mice. These data are consistent

with previous studies in clinical treatment of tumours found in the peritoneum showing the benefit of the IL-2 but also heterogeneity in the effects of treatment.51 The reason for this heterogeneity is not known, although it might reflect differences in the relative balance of effector cells and regulatory T cells.52 There is a great deal of interest in manipulating the immune response at specific sites exploiting the biological activity of cytokines. One innovative approach takes advantage of monoclonal antibodies to tumour-associated antigens (e.g. anti-HER-2/neu or anti-ganglioside GD2) that may have anti-tumour activities themselves, and genetically fuses them to cytokines (e.g. IL-2 or IL-12), which are then expressed and infused high throughput screening in vivo.53–55 Although the antibody fused to the cytokine diffuses throughout the recipient, it eventually accumulates at the tumour site as a result of antibody binding and retention so the cytokine concentration increases at tumour sites. This approach differs fundamentally

from the one presented in buy MG-132 the current work. In the current study the antibody does not bind the tumour but rather serves to inhibit the cytokine. The cytokine in the anti-tumour-associated antigen–antibody fusion is constitutively active and so may have unwanted effects. In contrast, in the approach demonstrated here, the cytokine activity is attenuated because of the specific binding component and increases only when activated by proteases. Another interesting strategy employs the latency-associated protein (LAP) of transforming growth factor-β (TGF-β) that is genetically fused to interferon-β (IFN-β) via a cleavable linker

recognized by an MMP such that the IFN-β becomes more active when the linker is cleaved. In this method, unlike the specific inhibition presented here, the LAP protein sterically shields the IFN-β from its receptor. This approach has been used to down-regulate inflammatory responses in a mouse model of arthritis.56 A variety of cytokines have been tested for their ability to act as adjuvants in the context of anti-tumour responses. Interestingly, while some studies found that immunization with irradiated Interleukin-2 receptor or mitomycin-treated transfected tumour cells expressing IL-2 can aid in initiating anti-tumour responses,57,58 other studies showed more modest effects.59 In contrast, viable tumour cells expressing even relatively low amounts of IL-2 within the tumour microenvironment can have dramatic immune effects and even result in tumour rejection.17,58,60,61 It is therefore likely that IL-2 produced by transfected growing tumours at the tumour site is largely acting locally, probably by enhancing T-cell and NK cell responses at the tumour site.

1D). This partial RING domain is insufficient to confer E3 ubiquitin ligase activity on viral Pellino since a recombinant form of the latter failed to catalyse the in vitro generation of polyubiquitin chains in the presence of E1 and E2 enzymes, whereas the mammalian member Pellino3S shows strong catalytic activity (Fig.

1E). Western immunoblotting using an anti-myc Hydroxychloroquine solubility dmso antibody shows that the lack of activity of viral Pellino relative to Pellino3 cannot be attributed to differences in protein quantity since both proteins show comparable levels of immunoreactivity. Interestingly, viral Pellino has a mobility corresponding to its predicted size of 25.4 kDa but it also shows a fainter immunoreactive band of slower electrophoretic mobility. The identity of this protein is unknown but its lack of reactivity with the anti-ubiquitin

antibody excludes NVP-BKM120 solubility dmso the possibility of the protein being modified by ubiquitination. The above analysis suggests that viral Pellino resembles its mammalian counterparts in containing a core FHA domain but differs in lacking both a wing appendage to the FHA domain and a functional RING-like motif. The emerging roles of Pellino proteins in TLR signalling coupled to the discovery of a viral homolog prompted studies on the ability of viral Pellino to regulate TLR signal transduction. Viral Pellino is encoded by the genome of MsEPV and given that the natural host of MsEPV is insect cells, the highly AT-rich sequence of the viral Pellino gene reflects an adaptation to this environment. In order

to ensure expression of viral Pellino in both insect and human cells, a form of the gene was chemically synthesised with codon sequences optimised for recognition by human translation machinery. This involved replacing As or Ts in the third position of each codon with a G or C, without altering the amino acid sequence of the translated protein. Such an approach was previously shown to enhance expression of poxviral genes in human cells 24. We initially MTMR9 assessed the effects of viral Pellino on Toll signalling in macrophage-like Drosophila S2 cells. A myc-tagged version of the viral protein showed uniform cytoplasmic distribution after transfection in these cells (Fig. 2A). The effects of increasing levels of viral Pellino expression on signalling by the Toll ligand C-106 was then assessed (Fig. 2B). C-106 is the active C-terminal fragment of the Spätzle protein and induced activation of a firefly luciferase reporter under the control of the drosomycin promoter. Toll signalling can induce expression of this antimicrobial peptide through the Rel family transactivators Dorsal and Dif. Thus, the activation of the drosomycin promoter was an especially relevant readout for Toll signalling in the present studies in light of the demonstration that Drosophila Pellino plays a key role in driving expression of drosomycin 13.

05) (data not shown). Host genetic factors are

proposed to be governing the pathology of HCV disease progression or regression along with the viral and environmental factors. Interplay of HLA-restricted T lymphocytes, antibody-secreting B lymphocytes, natural killer cells and cytokines conditions the immune response to viral infections. Effective presentation of viral antigens to CD4+ T cells and CD8+ T cells by HLA Class II and Class I molecules, respectively, is the key regulation of optimum immune response against viral infection and further AZD1208 mouse dictates viral clearance or persistence [20]. The results of the present study demonstrated that HLA-A11 is the only HLA buy Ipatasertib Class I antigens that show statistical significant association with chronic HCV infection (P = 0.001, Pc = 0.021), suggesting that HLA-A11 antigen may be a susceptibility antigen for viral persistence and chronic liver disease in Egyptian patients infected with HCV. Although HLA-B12, HLA-B13, HLA-B17 and HLA-B40 were more frequent in patients (P = 0.02, P = 0.04, P = 0.04, P = 0.02, respectively) and HLA-A32 (P = 0.03) and HLA-B14 (P = 0.015) were more frequent in controls, the significance was lost after correction for multiple testing and no other HLA Class I antigens were

associated with chronic HCV infection in this

study. The associations between HLA Class I antigens and the outcome of HCV infection are extensively investigated in different ethnic populations such as Caucasian Americans and populations from Korea, Italy, Russia, Spain, Ireland, Saudi Arabia, Western India, Japan and Germany [21–37]. The earlier reported associations showed ethnic and geographical differences sometimes with contradictory results. While HLA-A11 is associated with HCV persistence in Ireland [14, 25] in agreement with the results of the present study, Phosphoglycerate kinase HLA-A*1101 showed stronger association with viral clearance both in Caucasians and African Americans [29]. HLA-A32 in populations from Western India [27] and HLA-B14 in Italy [22] are associated with HCV infection in contrast to our findings. On other hand, several studies failed to demonstrate an association between the outcome of HCV infection and HLA Class I antigens [34–36]. In Egyptian, association was reported between HLA-A28, HLA-A29, HLA-B14 and HCV infection, and HLA-B50(21) with viral clearance in two cases of the studied sera [17]. HLA-A28 and HLA-29 were not detected in patients with HCV infection of the present study; in the same time, HLA-B14 shows a trend with protection (OR = 0.1) and not susceptibility.