Basophil activation by mosquito extracts in patients with hypersensitivity to mosquito bites

13301甲第4333号博士（医学）金沢大学博士論文要旨Abstract 以下に掲載：Cancer Science 106(8) pp.965-971 2015. Wiley Publishing Asia Pty Ltd. 共著者：Yasuhisa Sakakibara, Taizo Wada, Masahiro Muraoka, Yusuke Matsuda, Tomoko Toma, Akihiro Yachi

Basophil activation by mosquito extracts in patients with hypersensitivity to mosquito bites

Hypersensitivity to mosquito bites (HMB) is a cutaneous disorder belonging to the group of Epstein-Barr virus (EBV)-associated T/natural killer (NK)-cell lymphoproliferative diseases, and is primarily mediated by EBV-infected NK cells. It is characterized by intense local skin reactions accompanied by general symptoms after mosquito bites, and infiltration of EBV-infected NK cells into the bite sites. However, the mechanisms underlying these reactions have not been fully examined. We recently described the activation of circulating basophils by mosquito extracts in vitro in a patient with HMB. To further investigate this finding, we studied four additional patients with HMB. All patients showed typical clinical features of HMB after mosquito bites and they had NK lymphocytosis and high peripheral blood EBV DNA loads. We found evidence of EBV infection in NK cells through in situ hybridization that detected EBV-encoded small RNA-1, and flow cytometry showed HLA-DR expression on almost all NK cells. Basophil activation tests with the extracts of epidemic mosquitoes Culex pipiens pallens and Aedes albopictus showed positive responses to one or both extracts in all samples from patients with HMB, suggesting the presence of mosquito antigen-specific IgE and its binding to basophils. In particular, the extract of Aedes albopictus was able to activate basophils in all available patient samples. These results indicate that basophils and/or mast cells activated by mosquito bites may be involved in initiation and development of severe skin reactions to mosquito bites in HMB. © 2015 The Authors

Down-regulation of CD5 expression on activated CD8+ T cells in familial hemophagocytic lymphohistiocytosis with perforin gene mutations

Hemophagocytic lymphohistiocytosis (HLH) is characterized by uncontrolled activation of T cells and macrophages with overproduction of cytokines. Familial HLH type 2 (FHL2) is the most common form of primary HLH and is caused by mutations in PRF1. We have recently described a significant increase in the subpopulation of CD8+ T cells with clonal expansion and CD5 down-regulation in Epstein-Barr virus associated-HLH, which represented a valuable tool for its diagnosis. However, this unusual phenotype of CD8+ T cells has not been investigated fully in patients with FHL2. We performed immunophenotypic analysis of peripheral blood and measured serum pro-inflammatory cytokines in five patients with FHL2. All patients showed significantly increased subpopulations of activated CD8+ T cells with down-regulation of CD5, which were negligible among normal controls. Analysis of T-cell receptor Vβ repertoire suggested the reactive and oligoclonal expansion of these cells. The proportion of the subset declined after successful treatment concomitant with reduction in the serum levels of cytokines in all patients except one who continued to have a high proportion of the subset and died. These findings suggest that down-regulation of CD5 on activated CD8+ T cells may serve as a useful marker of dysregulated T cell activation and proliferation in FHL2. © 2013 American Society for Histocompatibility and Immunogenetics

Clonal expansion of Epstein-Barr virus (EBV)-infected γδ T cells in patients with chronic active EBV disease and hydroa vacciniforme-like eruptions

Chronic active Epstein-Barr virus (EBV) disease (CAEBV) is a systemic EBV-positive lymphoproliferative disorder characterized by fever, lymphadenopathy, and splenomegaly. Patients with CAEBV may present with cutaneous symptoms, including hypersensitivity to mosquito bites and hydroa vacciniforme (HV)-like eruptions. HV is a rare photodermatosis characterized by vesicles and crust formation after exposure to sunlight, with onset in childhood, and is associated with latent EBV infection. While γδ T cells have recently been demonstrated to be the major EBV-infected cell population in HV, the immunophenotypic features of EBV-infected γδ T cells in CAEBV with HV-like eruptions or HV remain largely undetermined. We describe three patients with CAEBV whose γδ T cells were found to be the major cellular target of EBV. HV-like eruptions were observed in two of these patients. A clonally expanded subpopulation of γδ T cells that were highly activated and T cell receptor Vγ9- and Vδ2-positive cells was demonstrated in all patients. We also show that the clonally expanded γδ T cells infiltrated into the HV-like eruptions in one patient from whom skin biopsy specimens were available. These results suggest the pathogenic roles of clonally expanded γδ T cells infected by EBV in patients with CAEBV and HV-like eruptions. © 2012 The Japanese Society of Hematology

Flow cytometric analysis of skin blister fluid induced by mosquito bites in a patient with chronic active Epstein-Barr virus infection

金沢大学医薬保健研究域医学系In chronic active Epstein-Barr virus (EBV) infection (CAEBV), ectopic EBV infection has been described in T or natural killer (NK) cells. NK cell-type infection (NK-CAEBV) is characterized by large granular lymphocytosis, high IgE levels and unusual reactions to mosquito bites, including severe local skin reactions, fever and liver dysfunction. However, the mechanisms underlying these reactions remain undetermined. Herein, we describe a patient with NK-CAEBV whose blister fluid after mosquito bites was analyzed. The patient exhibited significant increases in the percentage of CD56+ NK cells in the fluid compared with a simple mosquito allergy, in which the majority of infiltrated cells were CD203c+ cells, indicating basophils and/or mast cells. His fluid also contained CD203c+ cells, and his circulating basophils were activated by mosquito extracts in vitro. These results suggest that CD203c+ cells as well as NK cells may play pathogenic roles in the severe skin reactions to mosquito bites in NK-CAEBV. © 2009 The Japanese Society of Hematology.出版者に照会中.2010年12月より全文公開予定

Current Status of Large Helical Device and Its Prospect for Deuterium Experiment

Achievement of reactor relevant plasma condition in Helical type magnetic devices and exploration in its related plasma physics and fusion engineering are the aim of the Large Helical Device (LHD) project. In the recent experiments on LHD, we have achieved ion-temperature of 8.1 keV at 1 × 1019 m−3 by the optimization of wall conditioning using long pulse discharge by Ion Cyclotron Heating (ICH). The electron temperature of 10 keV at 1.6 × 1019 m−3 was also achieved by the optimization of Electron Cyclotron Heating (ECH). For further improvement in plasma performance, the upgrade of the Large Helical Device (LHD), including the deuterium experiment, is planned. In this paper, the recent achievements on LHD and the upgrade of LHD are described

Neural circuit analysis of axons regenerated by facial–hypoglossal nerve cross-link surgery

Introduction: Several methods of nerve reconstruction for facial nerve palsy are known. Although the recently introduced method of “cross-linking” of the facial and hypoglossal nerves with a grafted nerve has proved efficacious, the underlying mechanism is unclear. Methods: In this study, we created an animal model with Wistar rats and analyzed the newly reconstructed neural circuit by anterograde and retrograde neural tracer methods. The saphenous nerve was harvested as a graft, and its double end-to-side neurorrhaphy with the facial and hypoglossal nerves with epineural windows was carried out under the microscope. After an appropriate interval, small amounts of fluoro-ruby or fluoro-emerald were injected into the animals and analyzed 5 days later by fluorescent microscopy (Anterograde experiment: fluoro-ruby into the hypoglossal nucleus at 5 weeks; retrograde experiment: fluoro-ruby into the distal facial nerve sheath and fluoro-emerald into the distal hypoglossal nerve sheath, both at two months.). Results: The labeled axons derived from the hypoglossal nucleus were observed passing through the grafted nerve to the facial nerve. On the other hand, retrogradely labeled neurons were observed at both the hypoglossal and facial nuclei with some double-labeled neurons, suggesting that collateral sprouting had occurred. Conclusions: We suggest that the newly constructed neural circuits we observed are conducive to the treatment of facial nerve palsy