In contrast to the defective responses to IL-6, the inhibitory effects of IL-10 on IL-17 production were similar in healthy volunteers or HIES patients, suggesting that STAT3 is redundant for IL-10 signalling leading to reduced IL-17 production. In conclusion, the present study demonstrates that patients with HIES have differential defects in IL-17 responses to the two main pathogens associated with the disease, S. aureus and C. albicans, and this is comparable with the clinical features

of this syndrome. In addition, the extent of the Th17 defect is due to the location of the STAT3 mutation, and is associated with the clinical phenotype in these patients. Furthermore, defective Th17 responses are a more sensitive marker of the disease in HIES patients than STAT3 mutations. M. G. N. was supported by a Vidi Grant of the Netherlands Organization for Scientific Research. These studies were supported by donations PS-341 mouse collected by one of the HIES patients. None declared. “
“The detection and identification of bacteria present in natural and industrial ecosystems is now entirely based on molecular systems that detect microbial RNA or DNA. Culture methods were abandoned, in the 1980s, because direct observations showed that <1% of the bacteria in these

systems grew on laboratory media. Culture methods comprise the backbone of the Food and Drug Administration-approved diagnostic systems used in hospital laboratories, with some molecular methods Selleckchem RGFP966 being approved for the detection of specific pathogens that are difficult to grow in vitro. In several medical specialties, the reaction to negative cultures in cases in which overt signs of infection clearly exist has produced a spreading skepticism concerning the sensitivity and accuracy of traditional culture methods. We summarize evidence from the field of orthopedic surgery, and from other medical specialties, that support the contention that culture techniques are

especially insensitive and inaccurate in the detection of chronic biofilm infections. We examine the plethora of molecular techniques Protein Tyrosine Kinase inhibitor that could replace cultures in the diagnosis of bacterial diseases, and we identify the new Ibis technique that is based on base ratios (not base sequences), as the molecular system most likely to fulfill the requirements of routine diagnosis in orthopedic surgery. Biofilm infections were defined by Costertonet al. (1999), in a review in science, and were seen to encompass all device-related infections and a significant proportion of other chronic bacterial diseases. The characterization of an infection as being a biofilm infection is universally based on the unequivocal demonstration, by direct microscopy, of matrix-enclosed microbial communities within or upon the affected tissues or prostheses (Stoodleyet al., 2002).

41–43 Although some viral check details infections during pregnancy may be asymptomatic, approximately half of all preterm deliveries are associated with histologic evidence of inflammation of the placenta, termed acute chorioamnionitis (ACA)44 or chronic chorioamnionitis. Despite the high incidence of ACA, only a fraction of fetuses

have demonstrable infection. Most viral infections affecting the mother do not cause congenital fetal infection, suggesting that the placenta may play an important role as a potent immune-regulatory interface protecting the fetus from systemic infection.21,44 Recent observations indicate that the placenta functions as a regulator of the trafficking between the fetus and the mother rather than as a barrier.32 Fetal and maternal cells move in the two directions;45,46 similarly, some viruses and bacteria can reach the fetus by transplacental passage with adverse consequences. Although viral infections

are common during pregnancy, transplacental passage and fetal infection appear to be the exception rather than the rule. There is a paucity of evidence that viral infections lead to preterm Kinase Inhibitor Library cell assay labor; however, there are several areas of controversy and open questions. For example, what effects do subclinical viral infections of the decidua and/or placenta during early pregnancy have in response to other microorganisms such as bacteria? and what is the effect of a subclinical viral infection of the placenta on the fetus? Studies 3-oxoacyl-(acyl-carrier-protein) reductase from our laboratory suggest that the type of response initiated in the placenta may determine the immunological response of the mother and consequently, the pregnancy outcome. A placental infection that is able to elicit the production of inflammatory

cytokines, such as TNFα, INFγ, IL-12 and high levels of IL-6, will activate the maternal immune system and lead to placental damage and abortion or preterm labor.47 On the other hand, a viral infection in the placenta that triggers a mild inflammatory response will not terminate the pregnancy but might be able to activate the immune system, not only from the mother but also from the fetus as well. This activation may have several consequences: (1) sensitize the mother to other microorganisms, and therefore increase the apparent risk of pregnant women to infection; (2) promote an inflammatory response in the fetus, even though there is no viral transmission. Therefore it is critical to take into consideration that during pregnancy it is not only the maternal immune system responding, but also the fetal/placental unit. In the past, we have considered the placenta and fetus as non-active immunological organs which depend only on the action of the maternal immune system. Our data suggest the contrary. The placenta and the fetus represent an additional immunological organ which affects the global response of the mother to microbial infections. This is relevant for making decisions associated with treatment and prevention during pandemics.

epidermidis biofilms and the reduction in coverage was significant (P<0.001) for strains PAO1,

6750, 14:2, 23:1 and 27:1, but not for 15159. As for the dual-species biofilms shown in Fig. 3, a pronounced effect was seen for selleck chemicals llc strain 14:2. Similar effects were seen with the P. aeruginosa supernatants for the other S. epidermidis strains (Mia and C103), although the effects were less pronounced (data not shown). To determine whether the dispersal effect on S. epidermidis biofilms was due to cell lysis, S. epidermidis cells remaining in the biofilms after exposure to the P. aeruginosa biofilm supernatants were examined with the BacLight LIVE/DEAD stain. For all the S. epidermidis strains (Mia, C103 and C121), over 90% of the cells were viable after treatment

with each of the P. aeruginosa supernatants (data not shown). Protein Tyrosine Kinase inhibitor Similarly, the level of viability of the dispersed cells was over 90% as shown by staining or growth on 110 agar. In order to investigate what might be responsible for the variable effect of the P. aeruginosa strains (PAO1, NCTC 6750, 14:2, 23:1, 27:1 and 15159), biofilm supernatants were investigated for the release of a number of known virulence factors. The type strain PAO1 and the clinical isolate 15159 were found to be positive for the production of the quorum-sensing signal C4-HSL, while all the other strains were negative (Table 1). All the P. aeruginosa strains were positive for pyocyanin production, except 14:2 and 27:1, which were negative in this assay (Table 1). These results indicate that the repertoire of extracellular

products released from the 17-DMAG (Alvespimycin) HCl cells varies according to the strain. The secretion of extracellular proteases from P. aeruginosa cells growing in biofilms was investigated with zymography of culture supernatants (Fig. 5a). This showed differences between the strains in their degree of gelatinase activity. The supernatants from the two laboratory strains: PAO1 and NCTC 6750 as well as the clinical isolate 15159 contained at least three major bands of proteolytic activity at >150, 70 and 50 kDa. The >150 kDa enzyme has been identified previously by immuno-blotting and N-terminal sequencing as a multimeric form of P. aeruginosa elastase (Schmidtchen et al., 2003). In the same study, P. aeruginosa alkaline protease was demonstrated to band at around 50 kDa. This 50 kDa band, but not the higher molecular weight fractions, was also present in supernatants from strains 23:1 and 27:1 while the culture supernatant from biofilms of strain 14:2 appeared to lack any proteolytic activity. SDS-PAGE of the same material under reducing conditions confirmed differences in the extracellular protein profiles between the strains (Fig. 5b). Two different protein banding patterns could be identified, with strains PAO1, NCTC 6750 and 15159 showing a similar pattern and 14:2, 23:1 and 27:1 strains sharing many common bands.

At the systemic level, serum IgA peaked around 3 weeks post-infection (WPI) and decreased thereafter (Figure 3). Serum IgG quickly increased to a asymptote around three WPI and remained consistently high throughout the infection (Figure 3). Changes in serum IgA and IgG were significantly different between treatment

(infected and controls) and the interaction between treatment and WPI, PLX-4720 supplier when the analysis was corrected for the random effect of the host and the nonindependence of sampling the same individual over time (Table 4). Mucus IgA and IgG patterns exhibited similar trends: values were significantly higher in the infected, compared to controls, and decreased from section 1 to section 4 of the small intestine; mucus IgG also increased with sampling time in infected rabbits (Table 4). Graphidium strigosum: Infected rabbits mounted a strong somatic IgA and IgG response at the systemic level but the local antibody response was relatively low to both adult and L3 stages (Figures 4 and S2).

Specifically, serum IgA and IgG significantly differed between treatments and increased with infection time in infected individuals (Table 5). Mucus IgA and IgG were higher in the infected compared to the controls, and selleck chemicals for the infected, values increased with the course of infection showing stronger response in the fundus compared to the antrum section of the stomach (Table 5). Together these findings suggest that rabbits develop an effective systemic and local antibody response to T. retortaeformis but an inefficient mucosal response to G. strigosum. Trichostrongylus Tenofovir manufacturer retortaeformis: Total white blood cell

and lymphocyte counts were significantly higher in infected hosts compared to the controls and consistently increased over the course of the infection (Figure 5). No significant trend was recorded for eosinophils and neutrophils, corrected for the random effect of the host and the dependence of sampling the same individual over time (Figure 5). However, a more detailed analysis showed that during the second-to-fifth WPI, coinciding with the peak in antibody response, a strong eosinophilia, anaemia (haemoglobin) and high total white blood cells were recorded in infected compared to control rabbits (Table 6). Graphidium strigosum: A consistent increase in the concentration of eosinophils, lymphocytes, total white blood cells and haemoglobin was observed with the progression of the experiment but no significant differences were recorded between the infected and the controls (Figure 6, Table 6). In line with T. retortaeformis infection high eosinophilia, neutrophilia and total white blood cell concentrations were found during the second-to-the fourth WPI in infected compared to the controls; no significant development of anaemia was observed during this infection.

8–1.0 for overnight expression at 30°C) or 5 μg/mL soluble purified Fab. After washing, plates were incubated with HRP-conjugated/anti-human-Fab Ab. Detection was performed using TMB reagent (Sigma). For binding of peptide-loaded

Selleck Ulixertinib RTLs, ELISA plates were coated 2 h at 37°C with purified Fab, washed extensively and blocked for 30 min with PBS/2% skim milk. Loaded complexes were incubated for 1 h followed by 1 h incubation with anti-MHC-II mAb (TU39, BD pharmingen). After washing, plates were incubated with HRP-conjugated/anti-mouse-IgG Ab and detection was performed using TMB-reagent (Sigma). ELISA plates were coated with BSA-biotin and MHC-peptide complexes were immobilized as described in the Fab FLISA method above. Binding of soluble purified Fabs was performed by competitive-binding analysis, which examined the

ability of varied concentrations of soluble recombinant MHC-peptide complexes to inhibit the binding of the purified Fab to the specific immobilized MHC-peptide complex. Detection of Fabs binding to the immobilized MHC-peptide complexes was performed using TMB-reagent (Sigma). Cells were incubated for 4 h with medium containing 70 μM MOG-35-55 (MEVGWYRPPFSRVVHLYRNGK) or MBP-85-99 (ENPVVHFFKNIVTPR) for L-cell DR*1501 transfectants and with GAD-555-567 (NFFRMVISNPAAT) or control peptide: HA-307-319 Sirolimus datasheet (PKYVKQNTLKLAT), InsA-1-15 (GIVEQCCTSICSLYQ), and CII-261-273 (AGFKGEQGPKGEP)- for DR4-EBV-transformed B lymphoblast Preiss cells. Cells (106) were washed and incubated with 1–2 μg of specific Fab for 1 h at 4°C, followed by incubation with FITC-labeled anti-human Ab for 45 min at 4°C. Cells were finally washed and analyzed by a FACSCalibur flow cytometer (BD Biosciences). H2-1 T-cell hybridoma cells 51 (2×105/well in a 96-well plate) in 100 μL of 10% FBS-containing medium were combined with 2×105 irradiated (4500 rad) HLA-DRB1*1501-transfected L cells in 100 μL alone or in the presence of 10 μg/mL individual PRKACG peptides and incubated at 37°C

and 7% CO2 for 72 h. Supernatants were collected from the top of the culture, followed by centrifugation for 1 min at 1000 rpm. Hybridoma supernatants were added in triplicate into wells containing 5000 CTLL-2 cells in 100 μL of 10% FBS culture medium. After 24 h of culture, the cells were pulsed with 0.5 μCi [3H]thymidine for an additional 5 h and the net cpm (mean±SD) were calculated. Human MOG-35-55 peptide-specic H2-1 T-cell hybridoma cells (2×105/well) were co-cultured in triplicate with 2 mM Tris-containing medium alone, 8 μM RTL1000, or 8 μM RTL340 in 2 mM Tris-containing medium for 72 h. Aliquotted hybridoma cell cultures were thoroughly washed with RPMI and further stimulated with and without 10 μg/mL hMOG-35-55 peptide presented by irradiated (4500 rad) DRB1*1501-transfected cell lines at a 1:1 ratio in triplicate for 48 h.

In this connective tissue component, the orbit becomes inflamed, and infiltrated with T and B lymphocytes and mast cells [38]. The cytokines and disease-mediating factors generated by these infiltrating cells are currently thought to activate resident fibroblasts which exhibit a unique phenotype. Orbital fibroblasts comprise a heterogeneous population of cells, especially those derived from patients with TAO [39]. The cellular attributes peculiar to orbital fibroblasts are thought to underlie the susceptibility of the orbit to the manifestations of Graves’ disease. For instance, these fibroblasts exhibit particularly robust responses to proinflammatory cytokines such as the members of the IL-1 family.

When activated by IL-1β, leucoregulin or CD154, orbital selleckchem fibroblasts, especially those from patients with TAO, produce unusually high levels of hyaluronan [40]. This results from the induction of hyaluronan synthase (HAS) 1, 2 and 3

[41] and uridylyltransferase (UDP) glucose dehydrogenase [42]. The exaggerated induction of HAS isoforms could therefore account for the accumulation of hyaluronan in TAO. Activated orbital fibroblasts also express extremely high levels of IL-6, IL-8 and the prostaglandin endoperoxide H synthase-2, the inflammatory cyclooxygenase [43,44]. This latter induction, in turn, results in the production of extraordinarily high levels of prostaglandin E2 (PGE2) [45]. The prostanoid can exert an important bias on immune responses occurring in the orbit check details and favour T helper type 2 (Th2) predominance [46]. The magnitude of the induction of proinflammatory cytokines by orbital fibroblasts is remarkable but poorly understood. Cao and Smith reported the relatively low levels of secreted IL-1 receptor antagonist

(IL-1RA) produced by these cells [47]. Low levels of IL-1RA generation achieved following exposure to IL-1β results in poorly opposed IL-1α and IL-1β initiated signalling. Thus, Oxaprozin the amplitude of cytokine-provoked downstream gene expression is substantially greater than that achieved in other fibroblast types. The basis for the heterogeneity displayed by orbital fibroblasts is yet to be understood [48]. When sorted on the basis of whether or not they display Thy-1 (CD90), orbital fibroblasts can be categorized broadly as those possessing the potential to become adipocytes (Thy-1-) and those that can differentiate into myofibroblasts (Thy-1+) subsets [6]. Fibroblasts destined to become fat cells can do so spontaneously in culture or more efficiently when treated with prostacyclin together with compounds that increase intracellular cyclic adenosine-5′-monophosphate (cAMP) levels or with molecules that bind and activate PPAR γ[6,7]. Conversely, Thy-1+ fibroblasts differentiate into myofibroblasts that express high levels of smooth muscle actin. This occurs following their exposure to TGF-β.

The relapsing/remitting episodes of IBD 3 are associated with marked variations in pro-inflammatory cytokine production 4, 5; therefore, mouse models of IBD have been used to investigate the regulatory mechanisms that reduce inflammation and restore intestinal homeostasis 6. Dextran sodium sulfate (DSS)-induced colitis is a transient, myeloid-dependent gut injury model driven by epithelial cell damage 7. The severity of DSS colitis may be controlled by anti-inflammatory cytokines such as IL-10 and transforming growth

factor β (TGF-β) 8, but selleck products it is unclear whether these cytokines can directly modulate Mϕ function(s) in ways that promote the resolution of inflammation following the termination of DSS-induced injury 9–14. Furthermore, it is unknown whether IL-10 and TGF-β have redundant effects on Mϕ function 15, 16. TGF-β has multiple biological effects on hematopoietic and nonhematopoietic find more cells 17. Binding of TGF-β to TGF-βRII phosphorylates SMAD transcription factors that are primarily immunosuppressive in function 17. Genetic mutations in TGF-βRII are linked to UC and colitis-associated cancer in humans 18–20 and mice that lack TGF-β responsiveness in epithelial cells or T lymphocytes

develop severe intestinal inflammation 21, 22. Whether TGF-β suppresses colitic inflammation through direct effects on Mϕs is unknown. Herein, we employed the DSS colitis model to demonstrate that lack of TGF-β responsive Mϕs impairs the normal resolution of colitic inflammation. CD68TGF-βDNRII mice produce high levels of IL-33, an IL-1 family cytokine that is overexpressed in the colonic mucosa of UC patients 23–25. CD68TGF-βDNRII mice also produced significantly less IL-10 than littermate controls during colitis resolution. Taken together, these data show an important role for TGF-β in the specific regulation of intestinal Mϕ function in vivo. A transgenic Clomifene construct was generated to contain the human CD68 promoter (CD68-IVS1) 26, 27 followed by a human TGF-β receptor II lacking the cytoplasmic domain 28 (Fig. 1A). This truncated

receptor binds its extra-cellular ligand (TGF-β1, TGF-β2, and TGF-β3) but does not signal; therefore, it antagonizes TGF-β function in the cell by acting as a competitive inhibitor. This approach has been employed in a variety of tissue-specific promoter systems 21, 28–32. Pronuclear injection of C57BL/6 oocytes allowed generation of a founder (designated CD68TGF-βDNRII) possessing a single integration of approximately 15–20 copies (Fig. 1B). Thioglycollate-elicited peritoneal exudates cells (PECs) were evaluated by flow cytometry to determine the specificity of transgene expression. Compared with nontransgenic littermates, CD68TGF-βDNRII mice demonstrate TGF-βRII protein expression on CD11b+ myeloid cells (0.12 versus 5.3%), F4/80+ Mϕs (0.27 versus 7.9%), but not on CD11c+ dendritic cells (0.15 versus 0.32%), respectively (Fig. 1C).

[40, 41] The dependence of both human and murine macrophages on NO to control the pathogenesis of mycobacteria inside the host suggests that adequate activation of macrophages to produce this free radical is critical for host defence. In the present study, we demonstrated that IL-17A synergistically enhanced NO production and iNOS expression in BCG-infected macrophages in dose- and time-dependent manners. Kinetics study revealed that IL-17A enhanced iNOS expression at early time-points after BCG infection. Incubation of IL-17A did not further enhance iNOS expression in macrophages after 24 hr of BCG infection (Fig. 1c). Such observation can

be explained by negative feedback regulation on iNOS to prevent over-production of NO.[28, 29] Under the conditions we have tested, we observed that IL-17A

alone did not induce detectable levels of iNOS protein and NO production in macrophages. Our data suggest that IL-17A is able to prime Selleck NU7441 macrophages to produce NO in response to mycobacterial infection. Similar observations have been reported by Kawanokuchi et al.[42] – that IL-17A is able to enhance both iNOS expression and NO production in lipopolysaccharide-stimulated microglia, whereas IL-17A by itself has no effect on either product. In another study, IL-17A has been shown to induce iNOS expression and NO production in articular chondrocytes.[43] selleck kinase inhibitor Interleukin-17A also induces NO production in cartilage explants from osteoarthritis patients.[44] The differences between observations among these studies may implicate differential effects of IL-17A on NO production in specific cell types. Binding of the cell wall components (e.g. lipoarabinomannan and peptidoglycan) and secretory proteins (e.g. 38 000 molecular weight glycolipoprotein) of mycobacteria to Toll-like receptor 2 triggers the activation of multiple MAPKs in macrophages.[15, 45, 46] Consistent with our previous studies,[19, 21, 23] our results demonstrated that BCG is able to induce the phosphorylation of JNK, ERK1/2 and p38 MAPK and also translocation

of NF-κB p65 in macrophages. Our results revealed that IL-17A specifically enhanced BCG-induced phosphorylation of JNK in macrophages. Neither BCG-induced phosphorylation of ERK1/2 nor p38 MAPK was affected by IL-17A. Low-density-lipoprotein receptor kinase Moreover, our data suggest that the enhanced iNOS expression in IL-17A-pre-treated, BCG-infected macrophages can be explained by enhanced iNOS mRNA stability in these macrophages. Korhonen et al.[27] showed that cytokine-induced iNOS mRNA can be stabilized by a JNK signalling pathway through a tristetraprolin-dependent mechanism. The study may provide insights into the mechanism regarding our finding that IL-17A can enhance the stability of BCG-induced iNOS mRNA. Although our data indicate that NF-κB is not involved in IL-17A-enhanced iNOS expression in BCG-infected macrophages, other activated transcription factors may have been involved.

, 1990; Beggs, 1994). In vitro exposure of planktonic cells to amphotericin B often leads to a repression of ERG3 and ERG11 expression and a

concomitant decrease in ergosterol levels in the membrane, indicating that changes in the sterol composition are important for amphotericin B resistance in C. albicans (Liu et al., 2005). Furthermore, changes in the expression of genes involved in β-1,6-glucan Selleckchem FK866 biosynthesis (including SKN1 and KRE1) have also been proposed as a resistance mechanism against polyene antifungals (Gale, 1986; Mio et al., 1997; Liu et al., 2005). Antifungal resistance in C. albicans biofilms is a complex phenomenon, and like in planktonic cells, multiple mechanisms appear to be involved (Kuhn & Ghannoum, 2004). It was reported that efflux pumps are highly expressed in young biofilms (Ramage et al., 2002; Mukherjee et al., 2003; Mateus et al., 2004), even in the absence of an antifungal agent. However, the expression of genes encoding efflux pumps (CDR and MDR family) seems to be model system and/or strain dependent as CDR and MDR genes were not found to be overexpressed in the transcriptome studies of Garcia-Sanchez et al. (2004) and Murillo et al. (2005).

Nevertheless, some genes (including QDR1 and CDR4) appeared to be overexpressed in the study by Yeater et al. (2007) and other genes (including CDR2 at 12 h and MDR1 at 12 and at 24 h) were overexpressed in the in vivo model described by Nett et al. (2009). Reduced ergosterol levels (combined with see more increased levels of other sterols) also provide a possible resistance mechanism in biofilms (Mukherjee

et al., 2003) and changes in the expression levels of ERG genes were observed in several studies (Yeater et al., 2007; Nett et al., 2009). These changes probably lead to changes in the sterol composition of the cell membrane and may have a profound impact on antifungal resistance. Khot et al. (2006) and LaFleur et al. (2006) showed that resistant subpopulations (persisters) are present in C. albicans biofilms. Using untreated biofilms, Khot et al. (2006) compared the less-resistant, mafosfamide shear-removed, fraction of the biofilm with the basal blastospore subpopulation. In the latter, a marked downregulation of the ERG1 gene was observed, probably resulting in an overall downregulation of the ergosterol biosynthesis (remarkably, the expression of ERG11 was not altered). SKN1 and KRE1 were markedly upregulated in this resistant subpopulation. These changes in gene expression likely contributed to the observed amphotericin B resistance. When C. albicans biofilms in various stages of growth were treated with very high doses of fluconazole, an overexpression of genes involved in the ergosterol biosynthesis (ERG1, 3, 11 and 25) was observed, whereas after exposure to amphotericin B, an upregulation of SKN1 and KRE1 was observed. The transcriptional changes in sessile C.

Therefore, given

the Alarm Model, rejection would be expected in both situations. As this is not the case, the immune system of the F1 must have learned by a somatic process that its host expresses P1 and P2 epitopes and, therefore, must become tolerant of (unresponsive to) them, whereas the immune system of P1 is not tolerant of (is responsive to) P2 epitopes and vice versa because tolerance is epitope-specific. An adaptive immune system that Bortezomib nmr ignored the need to sort the repertoire as part of Decision 1 would lead to generalized autoimmunity because ‘perturbation’ cannot distinguish induction to responsiveness of anti-S from that of anti-NS cells. The activating Signal 2 must be NS-antigen-specific and, under the ARA Model, is delivered normally by eTh anti-NS that have undergone the sorting process. It is the presence or absence of Signal 2, not of costimulation, that distinguishes activation from inactivation. To argue that healthy tissues

induce tolerance whereas perturbed tissues induce responsiveness only begs the question as to how the epitopes of healthy and perturbed tissues are distinguished (i.e. how is epitope-specific tolerance established and maintained). Selleckchem Silmitasertib This is what the ARA Model attempts to do. Decision 1 is more meaningfully described as the sorting of the random repertoire. It is the resultant sorted repertoire, anti-NS, that normally faces the question of ‘whether to respond or not’ and with which effector ecosystem (i.e. Decision 2). Decision 2 is essentially a problem of the requirements for differentiation from a naïve/resting/initial state iT/B-cell anti-NS to an appropriate effector, eT/B anti-NS. Here, ‘perturbation’

is relevant. An Alarm Model for Decision 1 is irrelevant because the recognitive elements for alarm signals are germline-selected and antigen-unspecific. Decision 1 requires an individual-specific learning process that tells the immune system what is a host target (self) and that then uses this information to purge anti-self Dolichyl-phosphate-mannose-protein mannosyltransferase from the somatically generated random adaptive repertoire. By contrast, because the pathways used for Decision 2, the regulation of class, are germline-selected, the Alarm Model is clearly germane. The unique postulate of the Alarm Model is that the effector ecosystem induced ‘is tailored to the tissue……rather than to the invading pathogen’. This assumption is one of several possible alternatives under the Trauma Model that, as discussed above, can be tested by the proposed Experiment 2. Matzinger and Kamala give us a comprehensive model for tissue-based class control. It represents a heroic attempt to catalogue a vast number of observations into a form that can be put on an artist’s canvas (Figure 1 in ref. [30]).