These results suggest that the mannan within CMWS might be composed only of α-type mannose residues. For further structural characterization, we next analyzed the sample using NMR spectroscopy. Figure 4 shows the 1D-1H NMR spectra of CMWS. The spectrum of CMWS contained many
signals in the anomeric region of the mannose residues (δH 4.8–5.5 p.p.m.). Thus, we could not completely assign the signals using this technique. Therefore, we further examined samples using 1H, 13C-HSQC spectra to detect the number of signals from the mannose residues. Figure 5 shows the overlaid HSQC spectra of CMWS (black) and CAWS (blue). The overlaid HSQC spectra show 10 signals in the anomeric regions of their mannose residues (δH 4.8–5.5 p.p.m., δC 98–104 p.p.m.) that were arbitrarily labeled numbers 1–10 as described in Table 3. However, we could not completely assign all signals at this time. Therefore, we examined the anomeric www.selleckchem.com/products/AZD2281(Olaparib).html conformation of their carbohydrate residues because numerous studies have reported that the anomeric conformation of mannose residues is crucial R788 order for their pathogenicity and antigenicity (27, 28).
From the observed 1JH1,C1 obtained from 1H, 13C-HSQC spectra without decoupling during acquisition, all mannose residues were assigned to α-mannose (Table 3). We next examined samples using 2D TOCSY spectra to determine the linkage types of each residue according to the method of Shibata et al. (29). The findings are described in Table 3. Notably, no qualitative differences compared to CAWS were identified. In the present study, we clearly revealed that the CMWS, which is composed of a mannoprotein-β-glucan complex, dramatically induces coronary arteritis similar to that of KD, as well as acute anaphylactoid shock, in mice. These pathogenic effects are similar to those induced Cell press by CAWS. Moreover, the structure of mannan, which is considered a factor
in induction of the above-described pathogenicities, within CMWS was quite similar to that within CAWS. Based on these findings, we concluded that Candida mannan, especially α-mannan, might contribute to Candida pathogenicity with respect to coronary arteritis and acute shock. The CMWS used in this study was mainly composed of carbohydrates (mannose and glucose) and protein, with no endotoxin contamination (Table 1). Moreover, CMWS dramatically induced coronary arteritis (Figs 1 and 2) and acute anaphylactoid shock in mice (Table 2) in the same way as CAWS does (10–17). CMWS contains 50% carbohydrates and 10% proteins. Therefore, we attempted to further purify CMWS by dialysis. After dialysis, the carbohydrate content reached 80%, after which we again assessed its biological activity in terms of induction of vasculitis and acute anaphylactoid shock in mice. We found that this purified CMWS also exhibited both pathogenic effects on mice (data not shown).