Identification of Distinct Ligands for the C-type Lectin Receptors Mincle and Dectin-2 in the Pathogenic Fungus Malassezia

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Highlights? Mincle and Dectin-2 cooperatively recognize Malassezia through distinct ligands ? The Mincle ligands in Malassezia are glucosyl- and mannosylglycolipids ? The Dectin-2 ligand in Malassezia is an O-linked mannobiose-rich glycoprotein ? Cytokine response to the respective ligands was impaired in Mincle^?/? and Dectin-2^?/? DCs     

Identification and Multidimensional Optimization of an Asymmetric Bispecific IgG Antibody Mimicking the Function of Factor VIII Cofactor Activity

In hemophilia A, routine prophylaxis with exogenous factor VIII (FVIII) requires frequent intravenous injections and can lead to the development of anti-FVIII alloantibodies (FVIII inhibitors). To overcome these drawbacks, we screened asymmetric bispecific IgG antibodies to factor IXa (FIXa) and factor X (FX), mimicking the FVIII cofactor function. Since the therapeutic potential of the lead bispecific antibody was marginal, FVIII-mimetic activity was improved by modifying its binding properties to FIXa and FX, and the pharmacokinetics was improved by engineering the charge properties of the variable region. Difficulties in manufacturing the bispecific antibody were overcome by identifying a common light chain for the anti-FIXa and anti-FX heavy chains through framework/complementarity determining region shuffling, and by pI engineering of the two heavy chains to facilitate ion exchange chromatographic purification of the bispecific antibody from the mixture of byproducts. Engineering to overcome low solubility and deamidation was also performed. The multidimensionally optimized bispecific antibody hBS910 exhibited potent FVIII-mimetic activity in human FVIII-deficient plasma, and had a half-life of 3 weeks and high subcutaneous bioavailability in cynomolgus monkeys. Importantly, the activity of hBS910 was not affected by FVIII inhibitors, while anti-hBS910 antibodies did not inhibit FVIII activity, allowing the use of hBS910 without considering the development or presence of FVIII inhibitors. Furthermore, hBS910 could be purified on a large manufacturing scale and formulated into a subcutaneously injectable liquid formulation for clinical use. These features of hBS910 enable routine prophylaxis by subcutaneous delivery at a long dosing interval without considering the development or presence of FVIII inhibitors. We expect that hBS910 (investigational drug name: ACE910) will provide significant benefit for severe hemophilia A patients.     

Microtubule stabilization triggers the plus-end accumulation of Kif18A/kinesin-8

The precise control of spindle microtubule (MT) dynamics is essential for chromosome capture and alignment. Kif18A/kinesin-8, an essential regulator of kinetochore MT dynamics, accumulates at its plus-ends in metaphase but not prometaphase cells. The underlying mechanism of time-dependent and kinetochore MT-specific plus-end accumulation of Kif18A is unknown. Here, we examined the factors required for the MT plus-end accumulation of Kif18A. In Eg5 inhibitor-treated cells, Kif18A localized along the MTs in the monopolar spindle and rarely accumulated at their plus-ends, indicating that MT-kinetochore association was not sufficient to induce Kif18A accumulation. In contrast, taxol treatment triggered the rapid MT plus-end accumulation of Kif18A regardless of kinetochore association. Furthermore, Aurora B inhibitor-induced stabilization of the plus-ends of kinetochore MTs promoted the plus-end accumulation of Kif18A. In the absence of Kif18A, treatment with taxol but not Eg5 inhibitor causes highly elongated mitotic MTs, suggesting the importance of plus-end accumulation for the MT length-controlling activity of Kif18A. Taken together, we propose that there is a mutual regulation of kinetochore MT plus-end dynamics and Kif18A accumulation, which may contribute to the highly regulated and ordered changes in kinetochore MT dynamics during chromosome congression and oscillation.     

The antianxiety-like effect of astaxanthin extracted from Paracoccus carotinifaciens

Astaxanthin is a red carotenoid pigment and is widely found in living organisms. Astaxanthin has a potent antioxidative ability and has been reported as having various biological effects on the central nerve system, such as a protective effect against ischemia/reperfusion injury and improvement in cognitive function. In this study, to investigate the effects of astaxanthin on anxiety and depression, we performed some behavioral trials including the elevated plus maze test, hole-board test, forced swim test, and tail suspension test. Astaxanthin (100 and 300 mg/kg/day for 10 days, p.o.) significantly increased the time spent in open arms in the elevated plus maze test and increased the head-dipping count and duration in the hole-board test. On the other hand, astaxanthin (10, 100, 300, and 500 mg/kg/day for 10 days, p.o.) did not change the immobility time in the forced swim test or the tail suspension test. In conclusion, in mice, astaxanthin exerted anxiolytic-like effects, but not antidepressant-like effects.     

Distinct hemogenic potential of endothelial cells and CD41+ cells in mouse embryos

Definitive hematopoietic progenitor cells have been thought to develop from the vascular endothelium located in the aorta-gonad-mesonephros region of the mouse embryo. However, several recent findings have suggested that most hematopoietic progenitors are derived from non-endothelial precursor cells expressing CD41. We characterized two distinct precursor populations of definitive hematopoietic cell lineages, vascular endothelial (VE)-cadherin(+) CD41(-) CD45(-) endothelial cells and CD41(+) CD45(-) non-endothelial progenitors, both of which are derived from lateral mesoderm. VE-cadherin(+) endothelial cells obtained from cultures of differentiating embryonic stem cells possessed hematopoietic potential encompassing erythroid, myeloid and B lymphoid lineages, whereas CD41(+) progenitors lacked the B lymphopoietic potential. VE-cadherin(+) endothelial cells in the lower trunk of the embryo proper showed a significant potential for initiating B lymphopoiesis in cultures, while endothelial cells in the yolk sac appeared to have a bias for myeloerythropoietic differentiation. CD41(+) progenitors isolated from yolk sac and embryo proper were capable of generating multiple hematopoietic lineages, although mast cell precursors were exclusively enriched in CD41(+) progenitors in the yolk sac. These results suggest that hemogenic endothelial cells and CD41(+) progenitors possess distinct hematopoietic potential depending on the tissues in which they reside.     

Aspergillus takadae,a novel heterothallic species of Aspergillus section Fumigati isolated from soil in China

Aspergillus takadae is characterized by its heterothallic reproduction, pale yellow cleistothecia, broadly lenticular ascospores with two short equatorial crests and smooth convex surfaces, and broadly ellipsoidal to ovate conidia with a smooth wall. The validation of these novel species is supported further by the analyses of the β-tubulin, calmodulin, actin, and RPB2 sequences. In addition, the phylogenetic tree and DDBJ accession numbers of all the species of Aspergillus section Fumigati are presented. We report on the crossing of A. takadae species and the result of crossing A. takadae with a closely related species, A. spathulatus.     

Wetland environmental bioreactor system contributes to the decomposition of cellulose

Recently, numerous species of aquatic invertebrates inhabiting wetlands have been shown to possess endogenous cellulase, following the discovery that termites have cellulase genes encoded in their own genome rather than relying on symbiotic bacteria for decomposing cellulose. Wetlands have been empirically shown to play an important role in the decomposition of land‐originating hard‐to‐degrade polysaccharides such as cellulose. However, the mechanism that connects the cellulase producer and the wetlands remains unknown, which makes it very difficult to evaluate the ecological function of wetlands. Here we found that a macrobenthic bivalve, Corbicula japonica, secretes its cellulase to the wetland sediment. Secreted cellulases are immobilized in the components of the sediment. Moreover, adding cellulose or glucose to C. japonica could trigger its cellulase secretion level. These findings suggest a novel wetland cellulose decomposition mechanism. The decomposition ability of wetlands was previously ascribed only to microbes and/or invertebrates that contain cellulases. Our findings suggest that benthic animals supply wetlands with their enzymes as decomposition agents, while wetland sediments serve as immobilization scaffolds for the enzymes. This system, which was named by us an “environmental bioreactor system,” could provide a key function in wetlands.     

Homeostasis of the period of post-translational biochemical oscillators

Generally, circadian clocks or biological oscillations are resistant to external conditions such as temperature and nutrient concentration. We propose that enzyme-limited competition provides a general mechanism of homeostasis of the period of post-translational oscillators based on protein modifications, and demonstrate it by nutrient compensation in a theoretical model of cyanobacterial circadian clock. The rate change by nutrient concentration is counterbalanced by the amount of available free enzyme, which occurs because of the competition among the various substrates for the limited enzyme. The temperature and nutrient compensation are determined by the postulate that the catalytic modification reactions are rate limiting.     

The modes of action of colicins E5 and D,and related cytotoxic tRNases

Colicins E5 and D cleave the anticodon loops of distinct tRNAs of Escherichia coli both in vivo and in vitro, which accounts for their bactericidal actions through depletion of tRNAs and prevention of protein synthesis. The targets of colicin E5 are five tRNA species for four amino acids, tyrosine, histidine, asparagine and aspartic acid, and those of colicin D are four isoaccepting tRNAs for arginine. These two colicins represent a new class, the "tRNase-type", of the nuclease-type colicins, which previously comprised the DNase-type and ribotoxin-type (or rRNase-type). On the other hand, a certain clinical E. coli strain produces a potentially suicidal "anticodon-nuclease", PrrC, in response to phage T4 infection, which specifically cleaves its own lysine tRNA. For these three tRNases, i.e. colicins E5 and D, and PrrC, the substrates and reaction products, as well as their physiological consequences, are very similar to each other, but so many molecular features are different that these three proteins are assumed to have acquired similar functions through evolutionary convergence from different origins.     

The correlation among molecular phylogenetics, morphological data, and growth temperature of the genus Emericella, and a new species

The species of the genus Emericella have been classified and identified on the basis of morphological features. However, the phylogenetic relationships in this genus have not been investigated. To clarify the relationships according to molecular phylogenetics, morphological characteristics, and growth temperature regimens in Emericella, multilocus sequencing analysis based on recent Aspergillus taxonomy was carried out. Various characteristic species formed individual clades, and maximum growth temperature reflected the phylogenetics. Emericella species exhibit various ascospore characteristics, although some species do not have distinct ascospore ornamentation. Species that have smooth-walled ascospores with two equatorial crests are polyphyletic. Here, Emericella pachycristata is described and illustrated as a new species. Its ascospores are similar to those of E. nidulans. These species produce smooth-walled ascospores, but the equatorial crests of E. pachycristata are thicker than those of E. nidulans. On the phylogenetic trees, E. pachycristata is closely related to E. rugulosa, which produces ascospores with ribbed convex surfaces. Thus, E. pachycristata is considered to be a new species both morphologically and phylogenetically.