However, PF-02341066 in vivo it is now recognized that the chronic stimulation of this systemic inflammatory response provides markers for risk of disease, as well as the probability that the biomolecules of this response can actually contribute to the disease processes. Numerous studies have reported that chronic periodontal infections trigger chronic inflammation that is expressed locally as periodontitis [12,13], and systemically by elevations in various inflammatory mediators [2]. The levels of these mediators are associated generally with the severity/extent of periodontal disease, frequently decrease significantly with periodontal therapy and are decreased

in patients who become edentate (Cunningham LL, Novak MJ, Stevens J, Abadi B and Ebersole JL. The oral-systemic link: a bidirectional relationship. submitted.). Thus, while the ‘cause and effect’ relationship between the systemic inflammatory mediators and periodontitis is difficult to document unequivocally, the breadth of evidence indicates that chronic periodontal infections may be a contributor to the burden of risk for initiating and/or sustaining symptoms associated with chronic inflammatory diseases. We have described a non-human primate model of a chronic polymicrobial periodontal infection and have demonstrated a Napabucasin molecular weight pattern of host responses similar to those which occur in human disease

[53–55]. The baboon model of ligature-induced periodontitis and pregnancy can be used to assess the host response profiles during disease and to identify some biological links with adverse pregnancy outcomes [46]. Periodontitis in the non-human Endonuclease primates elicited by ligature placement is accompanied by changes in the subgingival microbial ecology with bacterial species similar to those in human disease [47,56,57]. This

chronic oral infection elicits elevated levels of local inflammatory, innate and acquired immune mediators [12,13,58,59]. The results of this report focused upon the capacity of the oral infection and disease to trigger changes in the systemic host response apparatus, manifested by changes in various acute phase reactants, and inflammatory mediators and cytokines/chemokines. Our previous results have demonstrated extensive variability in periodontal clinical presentation of the group of female baboons, not dissimilar from the heterogeneity reported in human populations, with some animals showing pre-existing naturally occurring mild to moderate periodontitis [46]. Additionally, while all the experimental animals subjected to tooth ligation developed significant increases in gingival inflammation and destructive disease following placement of ligatures during pregnancy, the changes in disease in response to ligation exhibited individual variation.

This was similarly seen in P. aeruginosa-infected cav1 KO mice [[9]]. Interestingly, IL-6 was also elevated not only in cav1 KO mice challenged with K. pneumoniae, but also in those exposed to P. aeruginosa. IL-6 plays disparate roles in inflammatory responses during bacterial infections [[25]]. IL-6 protects the host from death following K. pneumoniae infection; however, IL-6 neutralizing antibodies improve survival in

polymicrobial septic peritonitis [[26]]. Since IL-17R-deficient mice were shown to be more susceptible to H 89 in vitro K. pneumonia infection [[27]], we measured IL-17 levels and found an increase in cav1 KO mice compared with WT mice lungs. In fact, the susceptibility of IL-17-deficient mice to K. pneumoniae has been directly associated with delayed neutrophil recruitment and reduced G-CSF [[28]]. IL-17 has also been documented to induce secretion

of TNFα, IL-1β, and IL-6 [[29]]. The proinflammatory response to K. pneumoniae may not improve survival rates, but it aggravates existing disease conditions as shown in cav1 KO mice infected with P. aeruginosa [[9, 11]]. Despite the elevated levels of TNF-α, IL-1β, IL-6, and IL-17 in BAL fluid, the overall survival of cav1 KO mice with K. pneumoniae infection deteriorated rapidly. Interestingly, IL-27p28, a novel cytokine, was also increased in infected cav1 KO mice. p28, a subunit of IL-27, has broad inhibitory effects on Th1, Th2, and Th17 subsets

as well as the expansion of regulatory T cells [[30]]. Hence, we Rucaparib cost medroxyprogesterone propose that the elevated IL-27 may provide a passive regulatory mechanism during acute infection. Given that MIP2 is a chemokine primarily produced by macrophages, our finding that MIP2 levels were not elevated in the lung indicates an impaired alveolar macrophage population. This in turn suggests that distinct compartmental immunity occurs in K. pneumoniae infection [[31]]. In addition, the phagocytic ability of AMs was found to be downregulated in K. pneumoniae-infected cav1 KO mice (data not shown). It has been suggested that Cav1 is an immune-modulatory effector on cytokine production through the MKK3/p38 MAPK pathway [[32]]. We found that ERK1/2 was activated in cav1 KO mice. We also noted a decreased TLR-4 response that was previously linked to gram-negative bacteria, suggesting a troublesome lack of innate immunity in cav1 KO mice. We also observed that GSK3β−β-catenin−Akt pathway may be involved in this infection, with both Akt and β-catenin being downregulated by Cav1 deficiency. By contrast, GSK3β expression and phosphorylation are significantly increased following loss of Cav1. This is consistent with the previous studies that show that GSK3β can destabilize β-catenin [[17]]. Although Akt is usually an upstream signal for GSK3β [[33, 34]], in this case the Akt changes may result from the effects of GSK3β [[35]].

trachomatis-infected cells in vitro (Rasmussen et al., 1997). Still, the fact that increases in MICA are Gefitinib datasheet seen only on infected cells but not on uninfected bystanders in the same culture suggests that soluble mediators are not sufficient for these effects. Chlamydia trachomatis infection mediates MHC class I downregulation

through direct mechanisms involving the degradation of the transcription factor, RFX5, by chlamydia protease-like activity factor (Zhong et al., 2000). We have previously demonstrated that ‘soluble factors’ could also mediate the downregulation of MHC class I (Ibana et al., 2011a). The downregulation of MHC class I by cytokines, including IL-10 (Caspar-Bauguil et al., 2000) and CXCL12 (Wang et al., 2008) has been demonstrated in other PKC412 ic50 culture models, supporting our previous observation that MHC class I downregulation occurs indirectly in the bystander-noninfected cells present in C. trachomatis-infected A2EN cells (Ibana et al., 2011a). Cytokine-mediated induction of dendritic cell MICA transcription by IFNα has been reported (Jinushi et al., 2003), but the overall effects of cytokines on MICA expression appear to be quite pleiotropic with varying effects depending on cell

type and environment (reviewed in Champsaur & Lanier, 2010). In the present study, we observed that MICA is upregulated only in infected cells, demonstrating that the mechanisms underlying C. trachomatis-associated changes in MICA differ from those aminophylline altering expression of MHC class I and suggesting C. trachomatis infection does not promote the production of soluble MICA-inducing mediators in our culture system. MICA was first described as cell stress-induced protein in the gastrointestinal epithelium (Groh et al., 1996). Increased MICA expression has been observed during both viral (cytomegalovirus) and

bacterial (M. tuberculosis) infections (Groh et al., 2001; Das et al., 2001). Our observation that upregulation of MICA was limited to C. trachomatis-infected cells may indicate that this induction is via infection-derived stress or danger signals that are absent in noninfected bystander cells. Currently, the exact mechanism underlying the induction of MICA expression during viral and bacterial infection is not completely understood. Interestingly, a recent study suggested that human microRNAs can regulate MICA expression, allowing the maintenance of MICA protein expression at a particular threshold while facilitating acute upregulation of MICA during cellular stress (Stern-Ginossar et al., 2008). If C. trachomatis infection induces MICA expression by interfering with the host microRNA-mediated control pathways, this may explain why MICA induction does not occur on uninfected bystander cells. The latter effect would protect the host from unwarranted NK cell activation.

Networks are displayed graphically as gene/genes products (nodes) and the biological relationships between the nodes (edges). All edges are supported by at least one reference from

the literature, or from canonical information stored in the Ingenuity Pathways Knowledge Base. In addition, IPA computes a score for each network according to the fit of the user’s set of significant genes. The score, representing the – log(P-value), indicates the likelihood www.selleckchem.com/products/17-AAG(Geldanamycin).html of the Focus Genes in a network from Ingenuity Knowledge Database being found together randomly. We identified X chromosome sites that were consistently hypermethylated (n = 18, Table 1) and hypomethylated (n = 25, Table 2) in affected twins. Within the 5-kb window sampled for each X-linked gene, most of the differentially methylated regions (DMRs) were located in promoter regions or CpG islands while two hypermethylated (48980151–48980208 and 104355356–104355413) and six hypomethylated (103883076–103883125, selleck chemicals llc 47226194–47226247, 134532227–134532289, 134532327–134532376, 134532427–134532476, 134532627–134532676) DMRs were found downstream

of the transcription start sites. In all cases, DMRs were associated with known genes and we noticed that IL1RAPL2 was found in both lists (the two hypermethylated sites are downstream of the hypomethylated one). In some cases, multiple DMRs belong to the same gene (as in the case of hypomethylated peaks Resveratrol 134532227–134532289, 134532327–134532376, 134532427–134532476 and 134532627–134532676 onto gene DDX26B) or a specific site is located in a CpG island of

a bidirectional promoter for two different genes (i.e. hypomethylated peak 152712287–152712338 for genes SSR4 and IDH3G). Three hypomethylated peaks are associated with intergenic single-nucleotide polymorphisms (SNP), with peak 13087308–13087357, including SNP rs61677044, peak 13087708–13087757 falling in a region 150 bp downstream from SNP rs16978681, peaks 126140539–126140588 and 126140739–126140788 mapping to a SNP-rich region. Genes identified by the hypermethylated and hypomethylated sites encode for proteins that are illustrated in Tables 3 and 4, respectively. The 26 proteins include transcription factors, membrane and soluble enzymes, surface antigens and translocation proteins while in some cases proteins are currently defined only structurally, but not functionally. We explored possible functional relationships between the 26 genes using the IPA Knowledge Database. Unsupervised IPA network analysis identified a single cluster of 25 genes that included seven of our 26 genes and 18 additional genes, which was unlikely to occur by chance (P = 10−13). The plausible biological network generated is shown in Fig.

The BLT mouse has become widely used to study human immunobiology, and the findings presented here highlight important parameters for the generation of this model and its use. Overall, our data indicate that optimal human cell engraftment of BLT mice requires subrenal implant of thymic

tissues and low-dose irradiation. However, reasonable engraftment levels can be achieved in the absence of irradiation, and these BLT mice have an extended life span. Importantly, our study underscores the importance for considering click here the duration of experiments when using NSG–BLT mice, as these animals develop an activated human T cell population after 20 or more weeks post-implant in most cohorts. We thank Jamie Kady, Meghan Dolan, Pamela St Louis, Linda Paquin, Michael Bates, Bruce Gott, Allison Ingalls, Michelle Farley and Rebecca Riding for excellent technical assistance. This work was supported by National Institutes of Health FDA approved Drug Library order research grants AI046629 and DK032520, an institutional Diabetes Endocrinology Research Center (DERC) grant DK32520, a grant from the University

of Massachusetts Center for AIDS Research, P30 AI042845 and grants from the Juvenile Diabetes Research Foundation, International and the Helmsley Charitable Trust. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the National Institutes of Health. Michael A. Brehm is a consultant for The Jackson Laboratory. No other authors have conflicts of interest to declare. Fig. S1. Influence of the number of injected human CD34+ haematopoietic stem cells (HSC) on human cell chimerism in non-obese diabetic (NOD)-scid IL2rγnull- bone marrow, liver, thymus (NSG–BLT) mice. NSG mice were irradiated with 200 cGy (a,b)

or non-irradiated (c,d) were MG-132 manufacturer implanted with 1 mm3 fragments of human fetal thymus and liver in the renal subcapsular space and then injected intravenously with the indicated number of CD34+ HSC derived from the autologous human CD3-depleted fetal liver. The peripheral blood of recipient NSG mice was screened for human CD45+ cell chimerism (a,c) and development of human CD3+ T cells (b,d) at 12 weeks after implant. Each point shown represents an individual mouse. Fig. S2. Engraftment levels of human CD45+ cells in female or male non-obese diabetic (NOD)-scid IL2rγnull (NSG) mice implanted with tissues from either male or female donors. Male or female NSG mice were irradiated with 200 cGy, implanted with 1 mm3 fragments of human fetal thymus and liver in the renal subcapsular space and then injected intravenously with 1 × 105 to 5 × 105 CD34+ haematopoietic stem cells derived from the autologous human CD3-depleted fetal liver cells. Tissues both male (a) and female donors (b) were used. The peripheral blood of recipient NSG mice was screened for human CD45+ cell chimerism at 12 weeks after implant.

Comparisons with other Omp85s showed that the membrane domain is most conserved, whereas the periplasmic domain is more variable [23]. Interestingly, the protective activity of the Omp85 homolog, D15, from H. influenzae seemed to reside in the periplasmic part [21], suggesting that this part may be more important as a vaccine antigen. Possibly, the essential role of Omp85 in protein transport shields the membrane domain from interactions with the immune system. We used deoxycholate-extracted LY2157299 order OMVs for this study as

such vaccines are safe and efficacious in protection trials [2-4]. Most of the lipopolysaccharides (LPS), phospholipids and lipoproteins are removed by the detergent, which may possibly alter the conformation of Omp85 and other outer membrane antigens [53], as well as exposing epitopes that are masked in the Selleckchem Vismodegib native membrane. However, studies with native OMVs, in which outer membrane proteins are likely to be in their native conformation, support the notion that Omp85 is most probably non-bactericidal. Mice receiving such vaccines showed

negligible serum bactericidal activity against heterologous serogroup A strains [54], which also express Omp85 [5, 6, 17], as well as against some heterologous serogroup B strains [55]. It might be argued that the overexpressed Omp85 in the Omp85+ OMV vaccine in our study was not properly folded in the outer membrane and so explain why the increased Omp85 antibody levels did not result in higher bactericidal activities compared with the wt control vaccine. However, NMRI mice, immunized with the wt vaccine, showed equally high Omp85 antibody levels and serum bactericidal titres as the other mice strains given the Omp85+ vaccine (Figs. 2A and 3). As bactericidal antibodies are only induced by PorA in a native conformation [56, 57], the distinct bactericidal activity implied that the wt OMV vaccine expressed correctly folded PorA and presumably also

of Omp85. The negligible bactericidal activity of the NMRI sera with the PorA minus mutant showed that Omp85 antibodies did not contribute to the bactericidal activity. SBA with heterologous strains in other OMV vaccine studies also indicated that Omp85 antibodies were Glutamate dehydrogenase non-bactericidal [6, 7]. Neither did studies of Omp85 homologs from other bacteria demonstrate bactericidal activity of the specific antibodies although they were protective in animal models [18, 20]. To our knowledge, the only previous study of the vaccine potential of meningococcal Omp85 also found no bactericidal activity in OFI mice vaccinated with detergent-extracted OMVs containing Omp85 overexpressed by another genetic system [16]. Bactericidal activity was only demonstrated when the Omp85 sera were combined with sera following immunization with OMVs containing overexpressed levels of other minor OMPs.

While four other surface lipoproteins encoded on various cp32 plasmids (i.e. ErpG, ErpL, ErpX, and ErpY) have been shown to bind FH/FHL-1 from other animal sources, such as cattle, cat,

or dog (Stevenson et al., 2002), it is not clear what, if any, role this may play in the enzootic cycle of B. burgdorferi. In addition to the lipoproteins discussed in the preceding sections, there have also been several lipoproteins identified on the surface of B. burgdorferi that currently have no known function. Many of these were identified by Carroll and co-workers (i.e. lipoproteins PLX4032 price BBA65, BBA66, BBA71, and BBA73; Hughes et al., 2008) and through an examination of genes regulated by environmental cues through global expression profile analyses by Brooks et al. (Brooks et al., 2006; BBA689, BBA36, BBA66, BBA69, and BBI42). Given their cellular location on the surface, these lipoproteins likely perform an important role in either the tick or mammalian host environment, but future studies are needed to fully elucidate their functional role(s)

in B. burgdorferi virulence and/or Lyme disease pathogenesis. In addition to the numerous outer surface lipoproteins described previously, B. burgdorferi also contains integral OMPs that have transmembrane-spanning domains. OMPs are structurally different see more than lipoproteins in that they do not contain N-terminal lipid anchors. Bacterial OMPs, in general, provide an array of important functions, such as nutrient acquisition

(e.g. porins), antibiotic resistance (e.g. drug efflux pumps), protein transport and assembly, and cellular adhesion (Koebnik et al., 2000; Schulz, 2002; Bos et al., 2007). Likewise, B. burgdorferi OMPs also provide critical physiological functions for the spirochete cell, which is in accordance with the observation that nearly all known Etofibrate B. burgdorferi OMPs are encoded from stable chromosomal loci (Fraser et al., 1997). Interestingly, freeze-fracture electron microscopy has demonstrated that B. burgdorferi possesses a characteristically low abundance of integral OMPs, approximately 10-fold fewer than that detected in the Escherichia coli OM (Lugtenberg & van Alphen, 1983; Radolf et al., 1994). This paucity of integral membrane-spanning surface proteins, combined with the apparent limited antigenicity of OMPs, has seriously hindered identification of B. burgdorferi OMPs. As a result, relatively few nonlipoprotein surface proteins have been identified in B. burgdorferi, and even fewer have been fully characterized at the functional level. P66, encoded by ORF bb0603, was first identified as a 66-kDa chromosomally encoded B. burgdorferi antigen (Barbour et al., 1984; Coleman & Benach, 1987) with an immunogenic surface-exposed loop region (Bunikis et al., 1995, 1996; Probert et al., 1995).

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 Copanlisib order 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 Lumacaftor clinical trial 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 17-DMAG (Alvespimycin) HCl 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.

Table S1. Results from multiple linear regression fitting age and cytomegalovirus (CMV) status as co-variates. Table shows the unstandardized coefficient, significance and 95% confidence interval from the output of SPSS software for each CD45RA/CD27 subset. Unit of age is equal to 1 year. Table S2. Mean frequencies and the standard error of the mean of CD40 ligand (CD40L), interferon-γ (IFN-γ), interleukin-2 (IL-2) and tumour necrosis factor-α (TNF-α) in all possible combinations in each CD45RA/CD27 subset. “
“Hereditary angioedema (HAE) is a rare disease characterized by episodes of potentially

life-threatening angioedema. For affected children in the United Kingdom, there are relatively few data regarding disease prevalence, service organization and the humanistic burden of the disease. Decitabine cost To improve knowledge in these areas, we surveyed major providers of care for children with HAE. A questionnaire was sent to major paediatric centres to determine patient numbers, symptoms, diagnostic

difficulties, 5-Fluoracil datasheet management and available services. In addition, all patients at a single centre were given a questionnaire to determine the experiences of children and their families. Sixteen of 28 centres responded, caring for a total of 111 UK children. Seven children had experienced life-threatening crises. One-third of patients were on long-term prophylactic medication, including C1 inhibitor prophylaxis in four children. Eight centres reported patients who were initially misdiagnosed. Broad differences in management were noted, particularly regarding indications for long-term prophylaxis and treatment monitoring. We also noted substantial variation in the organization of services between centres, including the number of consultants contributing to patient care, Thiamet G the availability of specialist nurses, the availability of home therapy training and the provision of patient information. Ten of 12 patient/carer

questionnaires were returned, identifying three common themes: the need to access specialist knowledge, the importance of home therapy and concerns around the direct effect of angioedema on their life. To our knowledge, this study represents the first dedicated survey of paediatric HAE services in the United Kingdom and provides useful information to inform the optimization of services. “
“Galectin-3, an endogenous glycan-binding protein, plays essential roles during microbial infection by modulating innate and adaptive immunity. However, the role of galectin-3 within the CD4+CD25+Foxp3+ T regulatory (TREG) cell compartment has not yet been explored. Here, we found, in a model of Leishmania major infection, that galectin-3 deficiency increases the frequency of peripheral TREG cells both in draining lymph nodes (LNs) and sites of infection. These observations correlated with an increased severity of the disease, as shown by increased footpad swelling and parasite burden.

[25, 26] Candida spp., especially C. albicans, are able to produce and secrete various hydrolytic enzymes, particularly proteinases, lipases and phospholipases.[21] Shimizu et al. [27] and Abu-Elteen et al. [28] demonstrated the relevance of proteinases, hyaluronidases, condroitinases and phospholipases as virulence–related factors, reporting that secretory strains of Candida spp. showed an increased ability to invade tissues compared to non-secretory strains. According to Costa et al. [29], the activity of

proteinases and phospholipases is directly related to the promotion and establishment of infection. According to studies by Noumi et al. [30], hydrolytic enzymes and adhesins produced by C. albicans present themselves as the largest factor BMN 673 mouse associated with virulence, a fact previously suggested by Neugnot et al. [31]. Secreted aspartic proteinase (Sap) was first described in 1965 and was named Candida acid proteinase due to its optimal activity at acidic pH ranges and

because it was primarily found in yeast of the genus Candida.[32, 33] Sap may be considered Palbociclib clinical trial the most important hydrolytic enzyme among the virulence-associated factors of Candida spp.[34] Saps are believed to contribute to the adhesion and invasion of host tissues through the degradation or distortion of cell surface structures or the destruction of cells and molecules of the immune system, to avoid or resist microbicidal attack.[35, 36] Saps have a broad substrate specificity and are able to degrade a variety of human proteins such as albumin, haemoglobin, keratin, collagen, laminin, fibronectin, mucin and almost all immunoglobulins, including immunoglobulin A, which is resistant to the majority of bacterial proteinases.[37]

Basically, these enzymes are involved in the digestion of proteins by providing nitrogen to aid the survival of fungal cells.[38] At first glance, they appear to be acquiring nutrients; however, Saps may have developed other functions related to virulence such as degrading structural proteins tuclazepam and proteins of the immune system.[20, 21] In C. albicans, the production of Sap is encoded by a family of 10 SAP genes that are grouped into six subgroups or subfamilies: SAP1-3, SAP4-6, SAP7, SAP8, SAP9 and SAP10.[39-41] Gene transcription generates isoenzymes, named due to conformational and structural similarities among them.[40, 41] Sap1–Sap3 share 67% genetic identity and Sap4–Sap6 share as much as 89% identity. Sap1–Sap3 and Sap4–Sap6 are closely clustered. Sap7 only shares 20–27% identity with the other Sap proteins and is externally positioned. Sap8 is related to the clusters formed by Sap1–Sap3 and Sap4–Sap6. Sap9 and Sap10 differ from the other Sap1–8 isoenzymes and constitute a distinct group (Fig. 1).[42-44] All members of the family of Sap proteins possess four cysteine residues and two conserved aspartate residues.