A novel monoclonal antibody against 6-sulfo sialyl Lewis x glycans attenuates murine allergic rhinitis by suppressing Th2 immune responses

Abstract Lymphocyte homing is mediated by the interaction between L-selectin on lymphocytes and its glycoprotein ligands modified with 6-sulfo sialyl Lewis x (6-sulfo sLex) glycans on high endothelial venules (HEVs) in peripheral lymph nodes (PLNs). However, the lack of specific antibodies reactive with both human and mouse 6-sulfo sLex has limited our understanding of its function in vivo. Here, we generated a novel monoclonal antibody, termed SF1, that specifically reacts with 6-sulfo sLex expressed on HEVs in both species in a manner dependent on sulfate, fucose, and sialic acid modifications. Glycan array and biolayer interferometry analyses indicated that SF1 specifically bound to 6-sulfo sLex with a dissociation constant of 6.09 × 10–9 M. SF1 specifically bound to four glycoproteins from PLNs corresponding to the molecular sizes of L-selectin ligand glycoproteins. Consistently, SF1 inhibited L-selectin-dependent lymphocyte rolling on 6-sulfo sLex-expressing cells ex vivo and lymphocyte homing to PLNs and nasal-associated lymphoid tissues in vivo. Furthermore, SF1 significantly attenuated ovalbumin-induced allergic rhinitis in mice in association with significant suppression of Th2 immune responses. Collectively, these results suggest that SF1 can be useful for the functional analysis of 6-sulfo sLex and may potentially serve as a novel therapeutic agent against immune-related diseases

Role of MAdCAM-1-Expressing High Endothelial Venule-Like Vessels in Colitis Induced in Mice Lacking Sulfotransferases Catalyzing L-Selectin Ligand Biosynthesis

International audienc

Assembly of the draft genome of buckwheat and its applications in identifying agronomically useful genes.

世界初となるソバの全ゲノム解読に成功 －ソバの安全性、高品質性、収量安定性の鍵となる遺伝情報の発見－. 京都大学プレスリリース. 2016-04-13.Buckwheat (Fagopyrum esculentumMoench; 2n= 2x= 16) is a nutritionally dense annual crop widely grown in temperate zones. To accelerate molecular breeding programmes of this important crop, we generated a draft assembly of the buckwheat genome using short reads obtained by next-generation sequencing (NGS), and constructed the Buckwheat Genome DataBase. After assembling short reads, we determined 387, 594 scaffolds as the draft genome sequence (FES_r1.0). The total length of FES_r1.0 was 1, 177, 687, 305 bp, and the N50 of the scaffolds was 25, 109 bp. Gene prediction analysis revealed 286, 768 coding sequences (CDSs; FES_r1.0_cds) including those related to transposable elements. The total length of FES_r1.0_cds was 212, 917, 911 bp, and the N50 was 1, 101 bp. Of these, the functions of 35, 816 CDSs excluding those for transposable elements were annotated by BLAST analysis. To demonstrate the utility of the database, we conducted several test analyses using BLAST and keyword searches. Furthermore, we used the draft genome as a reference sequence for NGS-based markers, and successfully identified novel candidate genes controlling heteromorphic self-incompatibility of buckwheat. The database and draft genome sequence provide a valuable resource that can be used in efforts to develop buckwheat cultivars with superior agronomic traits

Photosensitized Protein-Damaging Activity, Cytotoxicity, and Antitumor Effects of P(V)porphyrins Using Long-Wavelength Visible Light through Electron Transfer

Photodynamic therapy (PDT) is a less-invasive treatment for cancer through the administration of less-toxic porphyrins and visible-light irradiation. Photosensitized damage of biomacromolecules through singlet oxygen (¹O₂) generation induces cancer cell death. However, a large quantity of porphyrin photosensitizer is required, and the treatment effect is restricted under a hypoxic cellular condition. Here we report the phototoxic activity of P­(V)­porphyrins: dichloroP­(V)­tetrakis­(4-methoxyphenyl)­porphyrin (CLP­(V)­TMPP), dimethoxyP­(V)­tetrakis­(4-methoxyphenyl)­porphyrin (MEP­(V)­TMPP), and diethyleneglycoxyP­(V)­tetrakis­(4-methoxyphenyl)­porphyrin (EGP­(V)­TMPP). These P­(V)­porphyrins damaged the tryptophan residue of human serum albumin (HSA) under the irradiation of long-wavelength visible light (>630 nm). This protein photodamage was barely inhibited by sodium azide, a quencher of ¹O₂. Fluorescence lifetimes of P­(V)­porphyrins with or without HSA and their redox potentials supported the electron-transfer-mediated oxidation of protein. The photocytotoxicity of these P­(V)­porphyrins to HeLa cells was also demonstrated. CLP­(V)­TMPP did not exhibit photocytotoxicity to HaCaT, a cultured human skin cell, and MEP­(V)­TMPP and EGP­(V)­TMPP did; however, cellular DNA damage was barely observed. In addition, a significant PDT effect of these P­(V) porphyrins on a mouse tumor model comparable with the traditional photosensitizer was also demonstrated. These findings suggest the cancer selectivity of these P­(V)­porphyrins and lower carcinogenic risk to normal cells. Electron-transfer-mediated oxidation of biomacromolecules by P­(V)­porphyrins using long-wavelength visible light should be advantageous for PDT of hypoxic tumor