Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Moriyama Masafumi Last modified date:2024.02.08

Professor / Division of Maxillofacial Diagnostic and Surgical Sciences / Department of Dental Science / Faculty of Dental Science


Papers
1. Ryoko Nagano, Shinsuke Fujii, Hiroko Wada, Mayu Matsumura-Kawashima, Yurie Mikami, Masafumi Moriyama, Toru Chikui, Kazunori Yoshiura, Seiji Nakamura, Tamotsu Kiyoshima, Lipomatous mixed tumor of the skin with cystic formation affecting the upper lip: A case report., Experimental and therapeutic medicine, 10.3892/etm.2022.11600, 24, 5, 664-664, 2022.11, Mixed tumor of the skin (MTS) is a rare neoplasm derived from the sweat glands with a reported frequency of 0.01-0.098% among all primary skin tumors. MTS often occurs in the head and neck region and is characterized by a mixture of epithelial, myoepithelial and stromal components. MTS also shows various morphological patterns, thus the presence of variants with rare components and its rarity make the clinical diagnosis even more difficult. A 47-year-old man was referred due to a painless, slowly growing, exophytic swelling intracutaneous mass of the upper lip. Magnetic resonance imaging revealed that the mass was a solid tumor with a fatty component in the proximal portion, while the distal portion was cystic and possibly contained highly viscous fluid. The mass was located between the skin and the orbicularis oris muscle in the upper lip. Excisional biopsy was performed and the lesion showed two intriguing features: A tumor with extensive lipomatous stroma and some large cysts. It was histopathologically diagnosed as lipomatous MTS with cystic formation in the upper lip. No evident signs of recurrence were observed during follow-up. The present report describes this case and includes a brief literature review of reported cases in the lip, since MTS can be confused with various skin lesions in clinical settings due to this rarity. Recognition by clinicians of different variants of MTSs, including the present case, is important for preventing erroneous diagnosis and treatment..
2. Kaneko N, Moriyama M, Maehara T, Chen H, Miyahara Y, Nakamura S., Orchestration of Immune Cells Contributes to Fibrosis in IgG4-Related Disease., Immuno, 10.3390/immuno2010013, 2, 1, 170-184, 2022.02.
3. Asami Nishikori, Midori Filiz Nishimura, Yoshito Nishimura, Kenji Notohara, Akira Satou, Masafumi Moriyama, Seiji Nakamura, Yasuharu Sato, Investigation of IgG4-positive cells in idiopathic multicentric Castleman disease and validation of the 2020 exclusion criteria for IgG4-related disease., Pathology international, 10.1111/pin.13185, 72, 1, 43-52, 2022.01, Patients with plasma cell type idiopathic multicentric Castleman disease (PC-iMCD) often show elevated serum IgG4 levels and IgG4-positive cell infiltration in tissues due to overproduction of interleukin-6, and may meet the diagnostic criteria for IgG4-related disease (IgG4-RD). Although PC-iMCD has been listed as a major exclusion disease for IgG4-RD, distinguishing between these diseases is challenging due to a lack of highly specific diagnostic biomarkers. In 2020, we proposed exclusion criteria of IgG4-RD mimickers. In this paper, we validated the accuracy of the criteria in excluding one of the mimickers, PC-iMCD, from IgG4-RD. Validation was performed on 57 PC-iMCD patients (39 presenting lymph node lesions and 19 with lung lesions) and 29 IgG4-RD patients (22 presenting lymph node lesions and seven with lung lesions). According to our results, 20.5% of the PC-iMCD patients with lymph node lesions and 42.1% of those with lung lesions met the diagnostic criteria for IgG4-RD. All these patients with PC-iMCD were excluded from a diagnosis of IgG4-RD by the proposed criteria. Additionally, 6.9% of IgG4-RD patients met the exclusion criteria. Thus, if the exclusion criteria are met, diagnosis should be made based on a combination of findings including organ distribution of disease, response to steroid therapy, and other pathological findings..
4. Akira Chinju, Masafumi Moriyama, Noriko Kakizoe-Ishiguro, Hu Chen, Yuka Miyahara, A S M Rafiul Haque, Katsuhiro Furusho, Mizuki Sakamoto, Kazuki Kai, Kotono Kibe, Sachiko Hatakeyama-Furukawa, Miho Ito-Ohta, Takashi Maehara, Seiji Nakamura, CD163+ M2 macrophages promote fibrosis in IgG4-related disease via TLR7/IRAK4/NF-κB signaling., Arthritis & rheumatology (Hoboken, N.J.), 10.1002/art.42043, 2021.12, OBJECTIVE: IgG4-related disease (IgG4-RD) is a fibro-inflammatory condition that can affect multiple organs. We previously demonstrated that human Toll-like receptor 7 (huTLR7)-transgenic C57BL/6 mice showed elevated serum IgG1 levels and inflammation with fibrosis in the salivary glands (SGs), lungs, and pancreas. Moreover, we observed extensive TLR7+ CD163+ M2 macrophage infiltration in SGs from IgG4-RD patients. Here, we examined the fibrotic mechanism via the TLR7 pathway. METHODS: Gene expression in SGs from huTLR7-transgenic mice and IgG4-RD patients was analyzed using DNA microarrays. We extracted the common upregulated TLR7-related genes in SGs from huTLR7-transgenic mice and IgG4-RD patients. Finally, we investigated the interaction between CD163+ M2 macrophages and fibroblasts before/after stimulation with the TLR7 agonist loxoribine. RESULTS: In huTLR7-transgenic mice and IgG4-RD patients, IL-1 receptor-associated kinase 3 (IRAK3) and IRAK4 were significantly overexpressed. Real-time PCR validated the upregulation of only IRAK4 in IgG4-RD patients compared with the other groups. IRAK4 was strongly detected in/around germinal centers in SGs from IgG4-DS patients alone. Double immunofluorescence staining showed that IRAK4-positive cells were mainly co-localized with CD163+ M2 macrophages in SGs. After stimulation with loxoribine, CD163+ M2 macrophages exhibited significantly enhanced expression of IRAK4 and nuclear factor kappa B (NF-κB) and increased supernatant concentrations of fibrotic cytokines. Finally, we confirmed that the number of fibroblasts was increased by culture with the supernatant of CD163+ M2 macrophages following stimulation with loxoribine. CONCLUSION: CD163+ M2 macrophages promote fibrosis in IgG4-RD by increasing the production of fibrotic cytokines via TLR7/IRAK4/NF-κB signaling. This article is protected by copyright. All rights reserved..
5. Kazuki Kai, Masafumi Moriyama, A S M Rafiul Haque, Taichi Hattori, Akira Chinju, Chen Hu, Keigo Kubota, Yuka Miyahara, Noriko Kakizoe-Ishiguro, Shintaro Kawano, Seiji Nakamura, Oral Squamous Cell Carcinoma Contributes to Differentiation of Monocyte-Derived Tumor-Associated Macrophages via PAI-1 and IL-8 Production., International journal of molecular sciences, 10.3390/ijms22179475, 22, 17, 2021.08, Tumor-associated macrophages (TAMs) promote cancer cell proliferation and metastasis, as well as anti-tumor immune suppression. Recent studies have shown that tumors enhance the recruitment and differentiation of TAMs, but the detailed mechanisms have not been clarified. We thus examined the influence of cancer cells on the differentiation of monocytes to TAM subsets, including CD163+, CD204+, and CD206+ cells, in oral squamous cell carcinoma (OSCC) using immunohistochemistry, flow cytometry, and a cytokine array. Furthermore, we investigated the effect of OSCC cells (HSC-2, SQUU-A, and SQUU-B cells) on the differentiation of purified CD14+ cells to TAM subsets. The localization patterns of CD163+, CD204+, and CD206+ in OSCC sections were quite different. The expression of CD206 on CD14+ cells was significantly increased after the co-culture with OSCC cell lines, while the expressions of CD163 and CD204 on CD14+ cells showed no change. High concentrations of plasminogen activator inhibitor-1 (PAI-1) and interleukin-8 (IL-8) were detected in the conditioned medium of OSCC cell lines. PAI-1 and IL-8 stimulated CD14+ cells to express CD206. Moreover, there were positive correlations among the numbers of CD206+, PAI-1+, and IL-8+ cells in OSCC sections. These results suggest that PAI-1 and IL-8 produced by OSCC contribute to the differentiation of monocytes to CD206+ TAMs..
6. Mizuki Sakamoto, Masafumi Moriyama, Mayumi Shimizu, Akira Chinju, Keita Mochizuki, Ryusuke Munemura, Keiko Ohyama, Takashi Maehara, Kenichi Ogata, Miho Ohta, Masaki Yamauchi, Noriko Ishiguro, Mayu Matsumura, Yukiko Ohyama, Tamotsu Kiyoshima, Seiji Nakamura, The diagnostic utility of submandibular gland sonography and labial salivary gland biopsy in IgG4-related dacryoadenitis and sialadenitis: Its potential application to the diagnostic criteria., Modern rheumatology, 10.1080/14397595.2019.1576271, 30, 2, 379-384, 2020.03, Objectives: In this study, we investigated the diagnostic utility of submandibular gland (SMG) sonography and labial salivary gland (LSG) biopsy as a less invasive procedure for diagnosing IgG4-related dacryoadenitis and sialadenitis (IgG4-DS)Methods: Sixty-eight patients with suspected IgG4-DS by presenting swelling of elevated serum IgG (>1747 mg/dl) and/or swelling glands underwent SMG sonography, LSG biopsy and measurement for serum IgG4. SMG sonographic diagnosis was determined by the following characteristic changes; 'hypoechoic areas of a nodal pattern with high vascularity' and/or 'hypoechoic areas of a reticular pattern in the superficial part'.Results: Thirty-one patients were diagnosed with IgG4-DS, 5 with IgG4-RD unaccompanied by lacrimal and salivary gland lesions, 28 with Sjögren's syndrome, and 4 with malignant lymphoma. The sensitivity, specificity, and accuracy of SMG sonography and LSG biopsy were 100%, 83.8%, 91.2% and 64.5%, 73.8%, 75.0%, respectively. Moreover, those of SMG sonography and LSG biopsy combined with serum IgG4 concentration (>135 mg/dl) were 100%, 94.6%, 97.1% and 64.5%, 91.9%, 79.4%, respectively.Conclusion: LSG biopsy needs to be extremely careful to diagnose IgG4-DS because of its low sensitivity. SMG sonography is sufficient for the diagnosis of IgG4-DS, especially when combined with serologic analysis. Thus, SMG sonography could adapt to the diagnostic criteria of IgG4-DS as a non-invasive method..
7. Yuko Ono, Hiroto Tsuboi, Masafumi Moriyama, Hiromitsu Asashima, Hanae Kudo, Hiroyuki Takahashi, Fumika Honda, Saori Abe, Yuya Kondo, Satoru Takahashi, Isao Matsumoto, Seiji Nakamura, Takayuki Sumida, RORγt antagonist improves Sjögren's syndrome‐like sialadenitis through downregulation of CD25, Oral Diseases, 10.1111/odi.13255, 26, 4, 766-777, 2020.03.
8. Takashi Maehara, Masafumi Moriyama, Seiji Nakamura, Review of a novel disease entity, immunoglobulin G4-related disease, Journal of the Korean Association of Oral and Maxillofacial Surgeons, 10.5125/jkaoms.2020.46.1.3, 46, 1, 3-11, 2020.02, Immunoglobulin G4 (IgG4)-related dacryoadenitis and sialoadenitis (IgG4-DS) are part of a multiorgan fibroinflammatory condition of unknown etiology termed IgG4-related disease (IgG4-RD), which has been recognized as a single diagnostic entity for less than 15 years. Histopathologic examination is critical for diagnosis of IgG4-RD. CD4+ T and B cells, including IgG4-expressing plasma cells, constitute the major inflammatory cell populations in IgG4-RD and are thought to cause organ damage and tissue fibrosis. Patients with IgG4-RD who have active, untreated disease exhibit significant increase of IgG4-secreting plasmablasts in the blood. Considerable insight into the immunologic mechanisms of IgG4-RD has been achieved in the last decade using novel molecular biology approaches, including next-generation and single-cell RNA sequencing. Exploring the interactions between CD4+ T cells and B lineage cells is critical for understanding the pathophysiology of IgG4-RD. Establishment of pathogenic T cell clones and identification of antigens specific to these clones constitutes the first steps in determining the pathogenesis of the disease. Herein, the clinical features and mechanistic insights regarding pathogenesis of IgG4-RD were reviewed..
9. Noriko Ishiguro, Masafumi Moriyama, Katsuhiro Furusho, Sachiko Furukawa, Takuma Shibata, Yusuke Murakami, Akira Chinju, A S M Rafiul Haque, Yuka Gion, Miho Ohta, Takashi Maehara, Akihiko Tanaka, Masaki Yamauchi, Mizuki Sakamoto, Keita Mochizuki, Yuko Ono, Jun-Nosuke Hayashida, Yasuharu Sato, Tamotsu Kiyoshima, Hidetaka Yamamoto, Kensuke Miyake, Seiji Nakamura, Activated M2 Macrophages Contribute to the Pathogenesis of IgG4-Related Disease via Toll-like Receptor 7/Interleukin-33 Signaling., Arthritis & rheumatology (Hoboken, N.J.), 10.1002/art.41052, 72, 1, 166-178, 2020.01, OBJECTIVE: IgG4-related disease (IgG4-RD) is a unique inflammatory disorder in which Th2 cytokines promote IgG4 production. In addition, recent studies have implicated the Toll-like receptor (TLR) pathway. This study was undertaken to examine the expression of TLRs in salivary glands (SGs) from patients with IgG4-RD. METHODS: SGs from 15 patients with IgG4-RD, 15 patients with Sjögren's syndrome (SS), 10 patients with chronic sialadenitis, and 10 healthy controls were examined histologically. TLR family gene expression (TLR-1 through TLR-10) was analyzed by DNA microarray in the submandibular glands (SMGs). Up-regulation of TLRs was confirmed in SGs from patients with IgG4-RD. Finally, the phenotype of human TLR-7 (huTLR-7)-transgenic C57BL/6 mice was assessed before and after stimulation with TLR agonist. RESULTS: In patients with IgG4-RD, TLR-4, TLR-7, TLR-8, and TLR-9 were overexpressed. Polymerase chain reaction validated the up-regulation of TLR-7 in IgG4-RD compared with the other groups. Immunohistochemical analysis confirmed strong infiltration of TLR-7-positive cells in the SGs of patients with IgG4-RD. Double immunohistochemical staining showed that TLR-7 expression colocalized with CD163+ M2 macrophages. After in vitro stimulation with a TLR-7 agonist, CD163+ M2 macrophages produced higher levels of interleukin-33 (IL-33), which is a Th2-activating cytokine. In huTLR-7-transgenic mice, the focus and fibrosis scores in SMGs, pancreas, and lungs were significantly higher than those in wild-type mice (P
10. Takashi Maehara, Ryusuke Munemura, Mayumi Shimizu, Noriko Kakizoe, Naoki Kaneko, Yuka Murakami, Moriyama Masafumi, Tamotsu Kiyoshima, Shintaro Kawano, Seiji Nakamura, Tissue-infiltrating immune cells contribute to understanding the pathogenesis of Kimura disease: A case report., Medicine, 10.1097/MD.0000000000018300, 98, 50, e18300, 2019.12, RATIONALE: Kimura disease (KD) is a rare, chronic inflammatory disorder characterized by subcutaneous granuloma in the head and neck region, as well as increased eosinophil counts and high serum immunoglobulin E (IgE) levels. Kimura disease is suspected to be an IgE-mediated disease, associated with an allergic response, in which antigen-specific B cells are stimulated to undergo specific IgE class switching with disease-specific CD4+ T (Th) cells help. Thus, exploration of the Th cells in affected tissues with KD is a highly promising field of the investigation. However, there have been no reports with direct evidence to implicate Th cells in affected lesions with KD. Here we quantitatively demonstrate that CD4+ GATA3+ T cells and interleukin (IL)-4+ IgE+ c-kit+ mast cells prominently infiltrate in affected lesion with KD. PATIENT CONCERNS: A 56-year-old Japanese man who exhibited painless swelling in the left parotid region. DIAGNOSES: Diagnosis of KD was made based on characteristic histopathologic findings, in conjunction with peripheral eosinophilia and elevated serum IgE levels. INTERVENTIONS: The patient underwent corticosteroid therapy and had been followed for 2 years. OUTCOMES: We report a rare case of KD of the parotid region in a 56-year-old man, followed by corticosteroid therapy for 2 years. The mass decreased in size and skin itchiness decreased after therapy. He was discharged without any complications. Furthermore, we quantitatively demonstrate the dominance of CD4+ GATA3+ T cells in affected tissues of KD and detect IL-4+ IgE+ c-kit+ mast cells in lesions by multicolor staining approaches. LESSONS: The findings from this case suggest that peripheral blood eosinophilia might serve as a marker of recurrent disease, long-term follow-up is necessary due to the possibility of recurrent. Interactions among expanded IgE+ B cells, CD4+ GATA3+ T cells, eosinophils, and activated mast cells might play a critical role in the pathogenesis of KD..
11. A S M Rafiul Haque, Masafumi Moriyama, Keigo Kubota, Noriko Ishiguro, Mizuki Sakamoto, Akira Chinju, Keita Mochizuki, Taiki Sakamoto, Naoki Kaneko, Ryusuke Munemura, Takashi Maehara, Akihiko Tanaka, Jun-Nosuke Hayashida, Shintaro Kawano, Tamotsu Kiyoshima, Seiji Nakamura, CD206+ tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production., Scientific reports, 10.1038/s41598-019-51149-1, 9, 1, 14611-14611, 2019.10, Tumor-associated macrophages (TAMs) promote tumor progression and inhibit anti-tumor immune response by producing various mediators and preferentially express CD163, CD204, and CD206. However, the role of these TAM subsets in oral squamous cell carcinoma (OSCC) remains unclear. Here we investigated the expression and function of TAM subsets in OSCC, especially in cancer cell proliferation. Biopsy sample from 44 patients with OSCC were examined for the expression of TAM markers and EGF by immunohistochemistry. EGF production of TAM subsets isolated from OSCC patients was assessed by flow cytometry. We also examined the effect of conditioned medium from TAM subsets on the proliferation of OSCC cells. CD163+ cells were detected diffusely all over the tumor and connective tissue area, while CD204+ and CD206+ cells were mainly detected in/around the tumors. Flow cytometric analysis found that CD206+ TAMs strongly produced EGF compared with CD163+ and CD204+ TAMs. Cell proliferation and invasion of OSCC cells cultured with conditioned medium of CD206+ TAMs were strongly enhanced and inhibited by anti-EGFR. The number of CD206+ TAMs positively correlated with worse clinical prognosis. Our results revealed differences in localization and EGF production among these TAM subsets. CD206+ TAMs might play a critical role in the proliferation of OSCC via EGF production..
12. Katsuhiro Furusho, Takuma Shibata, Ryota Sato, Ryutaro Fukui, Yuji Motoi, Yun Zhang, Shin-Ichiroh Saitoh, Takeshi Ichinohe, Masafumi Moriyama, Seiji Nakamura, Kensuke Miyake, Cytidine deaminase enables Toll-like receptor 8 activation by cytidine or its analogs., International immunology, 10.1093/intimm/dxy075, 31, 3, 167-173, 2019.03, Toll-like receptor 8 (TLR8), a sensor for pathogen-derived single-stranded RNA (ssRNA), binds to uridine (Uri) and ssRNA to induce defense responses. We here show that cytidine (Cyd) with ssRNA also activated TLR8 in peripheral blood leukocytes (PBLs) and a myeloid cell line U937, but not in an embryonic kidney cell line 293T. Cyd deaminase (CDA), an enzyme highly expressed in leukocytes, deaminates Cyd to Uri. CDA expression enabled TLR8 response to Cyd and ssRNA in 293T cells. CDA deficiency and a CDA inhibitor both reduced TLR8 responses to Cyd and ssRNA in U937. The CDA inhibitor also reduced PBL response to Cyd and ssRNA. A Cyd analogue, azacytidine, is used for the therapy of myelodysplastic syndrome and acute myeloid leukemia. Azacytidine with ssRNA induced tumor necrosis factor-α expression in U937 and PBLs in a manner dependent on CDA and TLR8. These results suggest that CDA enables TLR8 activation by Cyd or its analogues with ssRNA through deaminating activity. Nucleoside metabolism might impact TLR8 responses in a variety of situations such as the treatment with nucleoside analogues..
13. Mikami Y, Fujii S, Kohashi KI, Yamada Y, Moriyama M, Kawano S, Nakamura S, Oda Y, Kiyoshima T, Low-grade myofibroblastic sarcoma arising in the tip of the tongue with intravascular invasion: A case report., Oncology letters, 10.3892/ol.2018.9115, 16, 3, 3889-3894, 2018.09.
14. Haque Rafiul A.S.M., 森山 雅文, 久保田 恵吾, 石黒 乃理子, 坂本 瑞樹, 鎮守 晃, 望月 敬太, 坂本 泰基, 金子 直樹, 宗村 龍祐, 田中 昭彦, 前原 隆, 林田 淳之將, 川野 真太郎, 中村 誠司, CD206+腫瘍関連マクロファージは癌細胞増殖を促進する(CD206+ tumor-associated macrophages promote cancer cell proliferation), 日本口腔科学会雑誌, 67, 2, 103-103, 2018.07.
15. Sumida T, Azuma N, Moriyama M, Takahashi H, Asashima H, Honda F, Abe S, Ono Y, Hirota T, Hirata S, Tanaka Y, Shimizu T, Nakamura H, Kawakami A, Sano H, Ogawa Y, Tsubota K, Ryo K, Saito I, Tanaka A, Nakamura S, Takamura E, Tanaka M, Suzuki K, Takeuchi T, Yamakawa N, Mimori T, Ohta A, Nishiyama S, Yoshihara T, Suzuki Y, Kawano M, Tomiita M, Tsuboi H, Clinical practice guideline for Sjögren's syndrome 2017., Modern rheumatology, 10.1080/14397595.2018.1438093, 28, 3, 383-408, 2018.05.
16. Kenichi Ogata, Mayu Matsumura, Masafumi Moriyama, Wataru Katagiri, Hideharu Hibi, Seiji Nakamura, Cytokine Mixtures Mimicking Secretomes From Mesenchymal Stem Cells Improve Medication-Related Osteonecrosis of the Jaw in a Rat Model., JBMR plus, 10.1002/jbm4.10013, 2, 2, 69-80, 2018.03, Recently, several studies have demonstrated that intravenous administration of mesenchymal stem cells (MSCs) improve medication-related osteonecrosis of the jaw (MRONJ), and paracrine effects of secretomes from MSCs have been hypothesized as the primary contributors. These secretomes in conditioned media from human MSCs (MSC-CM) were previously demonstrated to promote bone and tissue regeneration. Because MSC-CM contain cytokines monocyte chemoattractant protein (MCP)-1, insulin growth factor (IGF)-1, and vascular endothelial growth factor (VEGF) at relatively higher concentrations than other factors, these cytokines were considered as relevant active factors for tissue regeneration. By mixing the recombinant proteins of MCP-1, IGF-1, and VEGF, included at the same concentrations in MSC-CM, we prepared cytokine mixtures mimicking MSC-CM and then evaluated its therapeutic effects in a rat MRONJ model. In vitro, cytokine mixtures promoted osteogenic differentiation, migration, and proliferation of rat MSCs. In addition, these maintained osteoclastic function. In vivo, we used a rat MRONJ model to examine therapeutic effects of the cytokine mixtures through intravenous administration. In MSC-CM or cytokine mixture group, open alveolar sockets in 66% or 67% of the rats with MRONJ, respectively, healed with complete soft tissue coverage and socket bones, whereas in the other groups, the exposed necrotic bone with inflamed soft tissue remained. Histological analysis revealed new bone formation and the appearance of osteoclasts in MSC-CM or cytokine mixture group; however, osteoclasts were significantly reduced in the other groups. Thus, we concluded that intravenous administration of cytokine mixtures might be an effective therapeutic modality for treating patients with MRONJ..
17. Y. Maruse, Shintarou Kawano, T. Jinno, Ryota Matsubara, Y. Goto, N. Kaneko, T. Sakamoto, Y. Hashiguchi, Masafumi Moriyama, T. Toyoshima, R. Kitamura, H. Tanaka, Kazunari Oobu, Tamotsu Kiyoshima, Seiji Nakamura, Significant association of increased PD-L1 and PD-1 expression with nodal metastasis and a poor prognosis in oral squamous cell carcinoma, International Journal of Oral and Maxillofacial Surgery, 10.1016/j.ijom.2018.01.004, 2018.01, Programmed cell death ligand 1 (PD-L1) and its receptor PD-1 are immune checkpoint molecules that attenuate the immune response. Blockade of PD-L1 enhances the immune response in a variety of tumours and thus serves as an effective anti-cancer treatment. However, the biological and prognostic roles of PD-L1/PD-1 signalling in oral squamous cell carcinoma (OSCC) remain to be elucidated. The purpose of this study was to examine the correlation of PD-L1/PD-1 signalling with the prognosis of OSCC patients to assess its potential therapeutic relevance. The expression of PD-L1 and of PD-1 was determined immunohistochemically in 97 patients with OSCC and the association of this expression with clinicopathological characteristics was examined. Increased expression of PD-L1 was found in 64.9% of OSCC cases and increased expression of PD-1 was found in 61.9%. Univariate and multivariate analysis revealed that increased expression of PD-L1 and PD-1 positively correlated with cervical lymph node metastasis. The expression of CD25, an activated T-cell marker, was negatively correlated with the labelling index of PD-L1 and PD-1. Moreover, the patient group with PD-L1-positive and PD-1-positive expression showed a more unfavourable prognosis than the group with PD-L1-negative and PD-1-negative expression. These data suggest that increased PD-L1 and PD-1 expression is predictive of nodal metastasis and a poor prognosis and is possibly involved in cancer progression via attenuating the immune response..
18. Takashi Maehara, Hamid Mattoo, Vinay S Mahajan, Samuel Jh Murphy, Grace J Yuen, Noriko Ishiguro, Miho Ohta, Masafumi Moriyama, Takako Saeki, Hidetaka Yamamoto, Masaki Yamauchi, Joe Daccache, Tamotsu Kiyoshima, Seiji Nakamura, John H Stone, Shiv Pillai, The expansion in lymphoid organs of IL-4+ BATF+ T follicular helper cells is linked to IgG4 class switching in vivo., Life science alliance, 10.26508/lsa.201800050, 1, 1, 2018.01, Distinct T follicular helper (TFH) subsets that influence specific class-switching events are assumed to exist, but the accumulation of isotype-specific TFH subsets in secondary lymphoid organs (SLOs) and tertiary lymphoid organs has not been hitherto demonstrated. IL-4-expressing TFH cells are surprisingly sparse in human SLOs. In contrast, in IgG4-related disease (IgG4-RD), a disorder characterized by polarized Ig class switching, most TFH cells in tertiary and SLOs make IL-4. Human IL-4+ TFH cells do not express GATA-3 but express nuclear BATF, and the transcriptomes of IL-4-secreting TFH cells differ from both PD1hi TFH cells that do not secrete IL-4 and IL-4-secreting non-TFH cells. Unlike IgG4-RD, IL-4+ TFH cells are rarely found in tertiary lymphoid organs in Sjögren's syndrome, a disorder in which IgG4 is not elevated. The proportion of CD4+IL-4+BATF+ T cells and CD4+IL-4+CXCR5+ T cells in IgG4-RD tissues correlates tightly with tissue IgG4 plasma cell numbers and plasma IgG4 levels in patients but not with the total plasma levels of other isotypes. These data describe a disease-related TFH subpopulation in human tertiary lymphoid organs and SLOs that is linked to IgG4 class switching..
19. Sachiko Furukawa, Kazunari Oobu, Masafumi Moriyama, Shintarou Kawano, Saori Sako, Hayashida Jun-Nosuke, Ryota Matsubara, Ken Ichi Ogata, Tamotsu Kiyoshima, Seiji Nakamura, Oral methotrexate-related lymphoproliferative disease presenting with severe osteonecrosis of the Jaw
A case report and literature review, Internal Medicine, 10.2169/internalmedicine.8946-17, 57, 4, 575-581, 2018.01, Long-term methotrexate (MTX) treatment can cause MTX-related lymphoproliferative disorder (MTX-LPD). We experienced a case of MTX-LPD that was associated with severe osteonecrosis of the jaw mimicking medication-related osteonecrosis of the jaw. The patient was an 81-year-old woman with rheumatoid arthritis (RA) who was treated with MTX and bisphosphonate. After 7 years, she was referred to our department for the assessment of giant ulcer and exposure of the alveolar bone of the left maxilla. Histopathological and immunological analyses confirmed a diagnosis of MTX-LPD. At seven months after the cessation of MTX treatment, the ulcerative and necrotic lesions had markedly decreased in size. A 1-year follow-up examination showed no evidence of recurrence and good RA control..
20. Tsuboi H, Hagiwara S, Asashima H, Takahashi H, Hirota T, Noma H, Umehara H, Kawakami A, Nakamura H, Sano H, Tsubota K, Ogawa Y, Takamura E, Saito I, Inoue H, Nakamura S, Moriyama M, Takeuchi T, Tanaka Y, Hirata S, Mimori T, Matsumoto I, Sumida T, Comparison of performance of the 2016 ACR-EULAR classification criteria for primary Sjögren's syndrome with other sets of criteria in Japanese patients., Annals of the rheumatic diseases, 10.1136/annrheumdis-2016-210758, 76, 12, 1980-1985, 2017.12.
21. H Takahashi, H Tsuboi, H Asashima, T Hirota, Y Kondo, M Moriyama, I Matsumoto, S Nakamura, T Sumida, cDNA microarray analysis identifies NR4A2 as a novel molecule involved in the pathogenesis of Sjögren's syndrome, Clinical and Experimental Immunology, 10.1111/cei.13000, 190, 1, 96-109, 2017.07, To examine genes expressed specifically in labial salivary glands (LSGs) of patients with Sjögren's syndrome (SS) in comparison with those of patients with immunoglobulin (Ig)G4-related disease (IgG4-RD), and to identify the genes involved in the pathogenesis of SS. Gene expression in LSGs of SS patients, IgG4-RD patients and healthy controls (HC) was analysed by cDNA microarray. Quantitative polymerase chain reaction (qPCR) was used to validate the up-regulation of differentially expressed genes (DEGs) in SS. Protein production of the validated gene in LSGs was examined by immunofluorescence (IF) assay. The association of molecular functions of the gene with the pathological conditions in SS was examined using peripheral blood lymphocytes. Among 1320 DEGs up-regulated in SS, qPCR confirmed the up-regulation of NR4A2 in LSGs of SS compared with IgG4-RD. IF staining showed higher production of NR4A2 in nuclei of CD4+ T cells and interleukin (IL)-17-producing cells in LSGs of SS, compared with IgG4-RD. Over-expression of NR4A2 mRNA was observed in peripheral CD4+ T cells of SS patients, compared with HC. Nuclear NR4A2 expression in T helper type 17 (Th17)-polarized CD4+ T cells determined by cellular IF was significantly higher in SS than in HC. Importazole, an inhibitor of importin-β, inhibited nuclear transport of NR4A2 and Th17 polarization along with IL-21 expression in naive CD4+ T cells under Th17-polarizing conditions, but did not alter retinoic acid receptor-related orphan receptor C (RORC) expression. NR4A2 seems to promote Th17 polarization via increased expression and intranuclear localization in CD4+ T cells of SS patients, which could play a critical role in the pathogenesis of SS..
22. Keigo Kubota, Masafumi Moriyama, Sachiko Furukawa, Haque A. S. M. Rafiul, Yasuyuki Maruse, Teppei Jinno, Akihiko Tanaka, Miho Ohta, Noriko Ishiguro, Masaaki Yamauchi, Mizuki Sakamoto, Takashi Maehara, Jun-Nosuke Hayashida, Shintaro Kawano, Tamotsu Kiyoshima, Seiji Nakamura, CD163(+) CD204(+) tumor-associated macrophages contribute to T cell regulation via interleukin-10 and PD-L1 production in oral squamous cell carcinoma, SCIENTIFIC REPORTS, 10.1038/s41598-017-01661-z, 7, 1, 1755, 2017.05, Tumor-associated macrophages (TAMs) promote cancer cell proliferation, invasion, and metastasis by producing various mediators. Although preclinical studies demonstrated that TAMs preferentially express CD163 and CD204, the TAM subsets in oral squamous cell carcinoma (OSCC) remain unknown. In this study, we examined the expression and role of TAM subsets in OSCC. Forty-six patients with OSCC were analyzed for expression of TAMs in biopsy samples by immunohistochemistry. We examined TAM subsets and their production of immune suppressive molecules (IL-10 and PD-L1) in peripheral blood mononuclear cells from three OSCC patients by flow cytometry. CD163 was detected around the tumor or connective tissue, while CD204 was detected in/ around the tumors. Flow cytometric analysis revealed that CD163(+) CD204(+) TAMs strongly produced IL-10 and PD-L1 in comparison with CD163+ CD204-and CD163-CD204+ TAMs. Furthermore, the number of activated CD3(+) T cells after co-culture with CD163+ CD204+ TAMs was significantly lower than that after co-culture with other TAM subsets. In clinical findings, the number of CD163+ CD204+ TAMs was negatively correlated with that of CD25+ cells and 5-year progression-free survival. These results suggest that CD163+ CD204+ TAMs possibly play a key role in the invasion and metastasis of OSCC by T-cell regulation via IL-10 and PD-L1 production..
23. Masaki Yamauchi, Masafumi Moriyama, Hayashida Jun-Nosuke, Takashi Maehara, Noriko Ishiguro, Keigo Kubota, Sachiko Furukawa, Miho Ohta, Mizuki Sakamoto, Akihiko Tanaka, Seiji Nakamura, Myeloid dendritic cells stimulated by thymic stromal lymphopoietin promote Th2 immune responses and the pathogenesis of oral lichen planus, PLoS One, 10.1371/journal.pone.0173017, 12, 3, 2017.03, Oral lichen planus (OLP) is a chronic inflammatory disease characterized by subepithelial Tcell infiltration. Recent studies reported that specific T helper (Th) subsets, especially Th2 cells, are involved in the pathogenesis of OLP. Thymic stromal lymphopoietin (TSLP) is mainly secreted by epithelial cells and potently activates myeloid dendritic cells (mDCs) to induce Th2-mediated inflammation. Here, we investigated the expression of TSLP and related molecules in OLP. Buccal mucosa specimens from patients with OLP, hyperkeratosis, and ulcer were analyzed by immunohistochemistry for expression of TSLP, its receptor (TSLPR), and inflammatory cells. TSLP was detected in/around the epithelium of patients with OLP and hyperkeratosis, whereas TSLPR, CD11c (mDC), and GATA3 (Th2) were strongly expressed in the subepithelial layer only in OLP patients. Double immunofluorescence staining showed that TSLPR expression mainly co-localized with CD11c. Moreover, the number of CD11c- and GATA-3 positive cells was correlated in OLP patients. In lesions selectively extracted by laser microdissection, the mRNA expression of Th2 (IL-4, MDC, TARC, GATA3)- and Th17 (IL-17, RORγt)-related molecules in OLP patients was significantly higher than in other groups. These results suggest that CD11c+ mDCs expressing TSLPR contribute to aberrant Th2 immune responses and the pathogenesis of OLP via TSLP stimulation..
24. Sachiko Furukawa, Masafumi Moriyama, Kensuke Miyake, Hitoshi Nakashima, Akihiko Tanaka, Takashi Maehara, Mana Iizuka-Koga, Hiroto Tsuboi, Jun-Nosuke Hayashida, Noriko Ishiguro, Masaki Yamauchi, Takayuki Sumida, Seiji Nakamura, Interleukin-33 produced by M2 macrophages and other immune cells contributes to Th2 immune reaction of IgG4-related disease, SCIENTIFIC REPORTS, 10.1038/srep42413, 7, 42413, 2017.02, IgG4-related disease (IgG4-RD) is characterized by elevated serum IgG4 and marked infiltration of IgG4-positive cells in multiple organs. Interleukin-33 (IL-33) is a recently described cytokine that is secreted by damaged epithelial cells, macrophages, and dendritic cells, and potently activates helper T type 2 (Th2) immune responses, which have been suggested to play a major role in IgG4 production of IgG4-RD. Here, we assessed the expression of IL-33 and related molecules in the salivary glands (SGs) of patients with IgG4-RD versus that in patients with Sjogren's syndrome (SS) and controls. Expression of IL-33 and its receptor (ST2) was strongly detected around ectopic germinal centers (GCs) in the SGs from patients with IgG4-RD, whereas IL-33 was expressed only in epithelial cells in patients with SS and controls. Moreover, IL-33 and CD68(+)/CD163(+) macrophages were mainly distributed around ectopic GCs in patients with IgG4-RD. Double immunofluorescence staining showed that IL-33 expression colocalized with CD68(+)/CD163(+) macrophages. Finally, mRNA expression levels of IL-33 showed a positive correlation to those of Th2 cytokines (IL-4 and IL-13) in patients with IgG4-RD. Our data suggest that IL-33 produced by M2 macrophages might contribute to the pathogenesis of IgG4-RD via aberrant activation of Th2 immune responses..
25. Maehara T, Mattoo H, Ohta M, Mahajan VS, Moriyama M, Yamauchi M, Drijvers J, Nakamura S, Stone JH, Pillai SS, Lesional CD4+ IFN-γ+ cytotoxic T lymphocytes in IgG4-related dacryoadenitis and sialoadenitis., Annals of the rheumatic diseases, 10.1136/annrheumdis-2016-209139, 76, 2, 377-385, 2017.02.
26. Masafumi Moriyama, Seiji Nakamura, Th1/Th2 immune balance and other T helper subsets in IgG4-related disease, Current Topics in Microbiology and Immunology, 10.1007/82_2016_40, 401, 75-83, 2017.01, IgG4-related disease (IgG4-RD) is a systemic disease characterized by elevated serum IgG4 levels and a strong infiltration of IgG4-positive plasma cells in various organs. IgG4-RD patients also frequently suffer from allergic diseases, including asthma and atopic dermatitis. It is well known that T helper type 2 (Th2) cells have an important role in the initiation of allergic diseases, and Th2 cytokines such as interleukin (IL)-4 and IL-13 promote class switching to IgG4. Therefore, IgG4-RD is considered to be a Th2-predominant disease. However, other Th subsets, including regulatory T cells and T follicular helper cells, have recently received increasing attention with regard to the pathogenesis of IgG4-RD. Exploring the interconnected network of Th subsets in IgG4-RD is a highly promising field of investigation. In this review, we focus on the localization and functions of individual Th subsets to clarify the involvement of these cells in the pathogenesis of IgG4-RD..
27. Masafumi Moriyama, Miho Ohta, Sachiko Furukawa, Yurie Mikami, Akihiko Tanaka, Takashi Maehara, Masaki Yamauchi, Noriko Ishiguro, Hayashida Jun-Nosuke, Shintarou Kawano, Yukiko Ohyama, Tamotsu Kiyoshima, Seiji Nakamura, The diagnostic utility of labial salivary gland biopsy in IgG4-related disease, Modern Rheumatology, 10.3109/14397595.2016.1148225, 26, 5, 725-729, 2016.09, Objective: For the definitive diagnosis of IgG4-related disease (IgG4-RD), biopsies of local lesions are recommended so as to exclude other diseases, including lymphoma and cancer. However, performing biopsies of underlying organs is technically difficult. In this study, we examined the diagnostic utility of labial salivary gland (LSG) biopsy as a less invasive procedure. Methods: Sixty-six patients with suspected IgG4-RD by clinical findings or high serum IgG4 underwent LSG biopsy. We examined the relationship between the number of IgG4-positive plasma cells in LSG and clinical findings. Results: The final diagnosis was 45 patients with IgG4-RD, 12 with Sjögren’s syndrome, four with suspected Sjögren’s syndrome, three with malignant lymphoma, one with systemic lupus erythematosus, and one with Warthin’s tumor. The sensitivity, specificity, and accuracy of LSG biopsy were 55.6%, 100.0%, and 70.0%, respectively. Forty-five IgG4-RD patients were divided into two groups: 1) 25 with lesions of salivary glands (IgG4-RD S+) and 2) 20 without these lesions (IgG4-RD S−). Seventeen of 25 (68.0%) IgG4-RD S + and 8 of 20 (40.0%) IgG4-RD S − patients were positive for LSG biopsy. In the IgG4-RD S − patients, the mean number of affected organs and serum IgG4 in the positive cases for LSG biopsy were significantly higher than in the negative cases. Conclusion: A solo LSG biopsy is insufficient for the diagnosis of IgG4-RD because of its low sensitivity. However, LSG biopsy combined with clinical findings, including serum IgG4 and number of affected organs, may contribute towards a diagnosis of IgG4-RD patients with affected underlying organs..
28. Yumi Imabayashi, Masafumi Moriyama, Toru Takeshita, Shinsuke Ieda, Jun-Nosuke Hayashida, Akihiko Tanaka, Takashi Maehara, Sachiko Furukawa, Miho Ohta, Keigo Kubota, Masaki Yamauchi, Noriko Ishiguro, Yoshihisa Yamashita, Seiji Nakamura, Molecular analysis of fungal populations in patients with oral candidiasis using next-generation sequencing, SCIENTIFIC REPORTS, 10.1038/srep28110, 6, 28110, 2016.06, Oral candidiasis is closely associated with changes in oral fungal biodiversity and is caused primarily by Candida albicans. However, the widespread use of empiric and prophylactic antifungal drugs has caused a shift in fungal biodiversity towards other Candida or yeast species. Recently, next-generation sequencing (NGS) has provided an improvement over conventional culture techniques, allowing rapid comprehensive analysis of oral fungal biodiversity. In this study, we used NGS to examine the oral fungal biodiversity of 27 patients with pseudomembranous oral candidiasis (POC) and 66 healthy controls. The total number of fungal species in patients with POC and healthy controls was 67 and 86, respectively. The copy number of total PCR products and the proportion of non-C. albicans, especially C. dubliniensis, in patients with POC, were higher than those in healthy controls. The detection patterns in patients with POC were similar to those in controls after antifungal treatment. Interestingly, the number of fungal species and the copy number of total PCR products in healthy controls increased with aging. These results suggest that high fungal biodiversity and aging might be involved in the pathogenesis of oral candidiasis. We therefore conclude that NGS is a useful technique for investigating oral candida infections..
29. Miho Ohta, Masafumi Moriyama, Takashi Maehara, Yuka Gion, Sachiko Furukawa, Akihiko Tanaka, Jun-Nosuke Hayashida, Masaki Yamauchi, Noriko Ishiguro, Yurie Mikami, Hiroto Tsuboi, Mana Iizuka-Koga, Shintaro Kawano, Yasuharu Sato, Tamotsu Kiyoshima, Takayuki Sumida, Seiji Nakamura, DNA Microarray Analysis of Submandibular Glands in IgG4-Related Disease Indicates a Role for MARCO and Other Innate Immune-Related Proteins, MEDICINE, 10.1097/MD.0000000000002853, 95, 7, e2853, 2016.02, IgG4-related disease (IgG4-RD) is a novel systemic disease entity characterized by elevated serum IgG4 and tissue infiltration of IgG4-positive plasma cells accompanied by severe fibrosis. Although recent studies demonstrated that innate immune cells including monocytes and macrophages might promote local fibrosis and IgG4 production, the pathological mechanism remains unclear. In this study, we sought to identify the disease-associated genes, especially innate immune molecules.
Gene expression was analyzed by DNA microarray in submandibular glands (SMGs) from patients with IgG4-RD (n=5), chronic sialoadenitis (CS) (n=3), and controls (n=3). Differentially expressed genes (DEGs) were validated by real-time polymerase chain reaction (PCR) and immunohistochemical staining in IgG4-RD (n=18), CS (n=4), Sjogren syndrome (n=11), and controls (n=10).
Gene expression patterns in the 3 groups were quite different from each other by the pvclust method and principal components analysis. In IgG4-RD, 1028 upregulated genes and 692 downregulated genes were identified as DEGs (PMARCO was identified as a disease-associated molecule in IgG4-RD by DNA microarray. Moreover, M2 macrophages might contribute to the initiation of IgG4-RD via MARCO..
30. Masafumi Moriyama, Seiji Nakamura, Potential pathways in the pathogenesis of IgG4-related disease, IgG4-Related Kidney Disease, 10.1007/978-4-431-55687-9_3, 43-54, 2016.01, IgG4-related disease (IgG4-RD) is a systemic disease characterized by elevated serum IgG4 and a marked infiltration of IgG4-positive plasma cells into affected organs. Regarding the immunological aspects of this disease, it is well known that IgG4 is induced by T helper type 2 (Th2) cytokines such as interleukin (IL)-4 and IL-13. Thus, IgG4-RD is considered to be a Th2-predominant disease. In addition, innate immune cells have recently received increasing attention with regards to the initiation of IgG4-RD. Exploring the mechanism of innate and acquired immunity in IgG4-RD is a highly promising field of investigation. In this chapter, we focus on the selective localization and functions of individual Th subsets and innate immune cells to clarify the contribution of these cells to the pathogenesis of IgG4-RD..
31. Masafumi Moriyama, Seiji Nakamura, Potential pathways in the pathogenesis of IgG4-related disease, IgG4-Related Kidney Disease, 10.1007/978-4-431-55687-9_3, 43-54, 2016.01, IgG4-related disease (IgG4-RD) is a systemic disease characterized by elevated serum IgG4 and a marked infiltration of IgG4-positive plasma cells into affected organs. Regarding the immunological aspects of this disease, it is well known that IgG4 is induced by T helper type 2 (Th2) cytokines such as interleukin (IL)-4 and IL-13. Thus, IgG4-RD is considered to be a Th2-predominant disease. In addition, innate immune cells have recently received increasing attention with regards to the initiation of IgG4-RD. Exploring the mechanism of innate and acquired immunity in IgG4-RD is a highly promising field of investigation. In this chapter, we focus on the selective localization and functions of individual Th subsets and innate immune cells to clarify the contribution of these cells to the pathogenesis of IgG4-RD..
32. Takahashi, Hiroyuki, Tsuboi, Hiroto, Iizuka, Mana, Asashima, Hiromitsu, Hirota, Tomoya, Kondo, Yuya, Matsumoto, Isao, Sumida, Takayuki, Nakamura, Seiji, Furukawa, Sachiko, Moriyama, Masafumi, Nakai, Yuji, Abe, Keiko, Yoshihara, Toshio, DNA Microarray Analysis of Labial Salivary Glands in Patients with Sjogren's Syndrome: Comparison with IgG4-Related Disease, ARTHRITIS & RHEUMATOLOGY, 67, 10, 2015.10.
33. Shimizu M, Okamura K, Kise Y, Takeshita Y, Furuhashi H, Weerawanich W, Moriyama M, Ohyama Y, Furukawa S, Nakamura S, Yoshiura K, Effectiveness of imaging modalities for screening IgG4-related dacryoadenitis and sialadenitis (Mikulicz's disease) and for differentiating it from Sjögren's syndrome (SS), with an emphasis on sonography., Arthritis research & therapy, 10.1186/s13075-015-0751-x, 17, 223, 2015.08.
34. Yoshikazu Hayashi, Masafumi Moriyama, Takashi Maehara, Yuichi Goto, Shintaro Kawano, Miho Ohta, Akihiko Tanaka, Sachiko Furukawa, Jun-Nosuke Hayashida, Tamotsu Kiyoshima, Mayumi Shimizu, Toru Chikui, Seiji Nakamura, A case of mantle cell lymphoma presenting as IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease, WORLD JOURNAL OF SURGICAL ONCOLOGY, 10.1186/s12957-015-0644-0, 13, 225, 2015.07, Background: Mantle cell lymphoma (MCL) is a relatively uncommon type of non-Hodgkin lymphoma. It develops in the outer edge of a lymph node called the mantle zone. In contrast, IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS) is characterized by elevated serum IgG4 and persistent bilateral enlargement of lacrimal glands (LGs) and salivary glands (SGs), with infiltration of IgG4-positive plasma cells. Recent studies indicated the importance of differentiation between IgG4-DS and malignant lymphoma.
Case presentation: An 82-year-old man was suspected of IgG4-DS because of a high serum IgG level (2174 mg/dL) and bilateral swelling of LGs and SGs. Lip biopsy and fine needle biopsy of submandibular gland were performed, and subsequently, MCL was diagnosed through the histopathological findings.
Conclusions: MCL most commonly occurs in the Waldeyer ring, but rarely in the stomach, spleen, skin, LG, and SG. We report an unusual case of MCL involving LGs and SGs mimicking IgG4-DS, which suggests that IgG4 testing may be useful in the differentiation of IgG4-DS in the presence of bilateral swelling of LGs or SGs..
35. Teppei Jinno, Shintaro Kawano, Yasuyuki Maruse, Ryota Matsubara, Yuichi Goto, Taiki Sakamoto, Yuma Hashiguchi, Naoki Kaneko, Hideaki Tanaka, Ryoji Kitamura, Takeshi Toyoshima, Akiko Jinno, Masafumi Moriyama, Kazunari Oobu, Tamotsu Kiyoshima, Seiji Nakamura, Increased expression of interleukin-6 predicts poor response to chemoradiotherapy and unfavorable prognosis in oral squamous cell carcinoma, ONCOLOGY REPORTS, 10.3892/or.2015.3838, 33, 5, 2161-2168, 2015.05, Recent studies have revealed that cancer cells are exacerbated by chronic inflammation. The present study examined the immunohistochemical expression for interleukin-6 (IL-6), a pleiotropic inflammatory cytokine, in oral squamous cell carcinoma (OSCC) to elucidate the association of IL-6 expression with tumor progression, chemoresistance and prognosis. Seventy-eight patients with primary OSCC were analyzed by immunohistochemical staining for IL-6. These labeling indexes (LIs) were calculated and evaluated in association with the clinicopathologic characteristics and prognosis in the OSCC patients. The patients were divided into three groups as follows: negative group = LI = 30%. The patient numbers of the negative, low and high expression groups were 24, 22 and 32, respectively. In the high IL-6 expression group, IL-6 receptor (IL-6R), phospho-signal tranducer and activator of transcription 3 (p-STAT3) were also detected in almost all the cancer cells. The prevalence of the cervical lymph node or the distant metastasis in the high expression group was significantly higher than those in the negative and low expression groups. Furthermore, the high expression group had a significantly poorer tumor response to the preoperative chemoradiotherapy and a more unfavourable prognosis than the negative and the low expression groups. Interestingly, IL-6, IL-6R and p-STAT3 were expressed in the residual cancer cells of all the patients in the high expression group with poor response to chemoradiotherapy. These results suggested that IL-6 signaling possibly is involved in the progression and treatment-resistance of OSCC and IL-6 expression in cancer cells could be a useful predictive factor of poor response to chemoradiotherapy and unfavorable prognosis..
36. Takahashi, Hiroyuki, Tsuboi, Hiroto, Nakai, Yuji, Iizuka, Mana, Asashima, Hiromitsu, Hirota, Tomoya, Kondo, Yuya, Moriyama, Masafumi, Furukawa, Sachiko, Matsumoto, Isao, Nakamura, Seiji, Yoshihara, Toshio, Abe, Keiko, Sumida, Takayuki, DNA Microarray Analysis of Labial Salivary Glands in Patients with Sjogren's Syndrome: Comparison with IgG4-Related Disease, SCANDINAVIAN JOURNAL OF IMMUNOLOGY, 81, 5, 420-420, 2015.05.
37. IL-33 Secreted by M2 Macrophage Promotes the Pathogenesis of IgG4-Related Disease.
38. Takashi Maehara, Masafumi Moriyama, Shintaro Kawano, Jun-Nosuke Hayashida, Sachiko Furukawa, Miho Ohta, Akihiko Tanaka, Masaki Yamauchi, Yukiko Ohyama, Tamotsu Kiyoshima, Seiji Nakamura, Cytokine Profiles Contribute to Understanding the Pathogenic Difference Between Good Syndrome and Oral Lichen Planus Two Case Reports and Literature Review, MEDICINE, 10.1097/MD.0000000000000704, 94, 14, e704, 2015.04, We described and analyzed the pathogenic difference between Good syndrome (GS) and oral lichen planus (OLP) in oral mucosa.
Good syndrome (GS) is a rare disease characterized by B and T cell immunodeficiency associated with hypogammaglobulinemia and thymoma. GS patients frequently develop oral lichenoid lesions with lymphocytic infiltration beneath the basal layer. Oral lichen planus (OLP) is a chronic inflammatory disease of the oral mucosa characterized by destruction of basal cells by Langerhans cells, macrophages, and T lymphocytes. Although the histological features of the lesions of both diseases are very similar, the pathogenesis of GS in the oral mucosa remains unknown. In this study, we thus investigated the expression of infiltrating lymphocyte subsets (CD3, CD20, CD4, and CD8) and T helper (Th) cytokines including interferon (IFN)-g (Th1 type), interleukin (IL) -4 (Th2 type), IL -17 (Th17 type), and IL -10 (regulatory T cell type) by immunohistochemistry in buccal mucosa specimens from 2 GS patients compared with 15 OLP patients. All patients showed a predominance of CD3(+) T cells over CD20(+) B cells, and CD4(+) Th cells over CD8(+) cytotoxic T cells. This polarization was especially prominent in GS. IFN-gamma and IL-10 were strongly detected in the infiltrating lymphocytes of all patients. However, IL -4 and IL -17 were detected in OLP patients only.
These results suggest that the pathogenesis of GS is different from that of OLP. GS is a unique inflammatory disorder characterized by dysfunction of Th2 and Th17 immune reactions via abnormal T -B cell interaction..
39. S. Furukawa, Masafumi Moriyama, Shintarou Kawano, A. Tanaka, T. Maehara, Hayashida Jun-Nosuke, Y. Goto, Tamotsu Kiyoshima, H. Shiratsuchi, Yukiko Ohyama, M. Ohta, Y. Imabayashi, Seiji Nakamura, Clinical relevance of Küttner tumour and IgG4-related dacryoadenitis and sialoadenitis, Oral Diseases, 10.1111/odi.12259, 21, 2, 257-262, 2015.03, Objectives: Küttner tumour (KT), so-called chronic sclerosing sialoadenitis, is characterised by concomitant swelling of the submandibular glands secondary to strong lymphocytic infiltration and fibrosis independent of sialolith formation. However, recent studies have indicated that some patients with KT develop high serum levels of IgG4 and infiltration of IgG4-positive plasma cells, namely IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS), so-called Mikulicz's disease. The aim of this study was to clarify the clinical and pathological associations between KT and IgG4-DS. Materials and Methods: Fifty-four patients pathologically diagnosed with KT or chronic sialoadenitis were divided into two groups according to the presence or absence of sialolith (KT-S (+) or KT-S (-), respectively). Results: There were no significant differences in the clinical findings, including the mean age, sex and disease duration, between the two groups. All patients in the KT-S (+) group showed unilateral swelling without infiltration of IgG4-positive plasma cells or a history of other IgG4-related diseases (IgG4-RD), while those in the KT-S (-) group showed bilateral swelling (37.5%), strong infiltration of IgG4-positive plasma cells (87.5%) and a history of other IgG4-RD (12.5%). Conclusions: These results suggest an association between the pathogeneses of KT-S (-) and IgG4-DS, but not KT-S (+)..
40. Keiko Oyama, Masafumi Moriyama, Hayashida Jun-Nosuke, A. Tanaka, T. Maehara, S. Ieda, S. Furukawa, M. Ohta, Y. Imabayashi, Seiji Nakamura, Saliva as a potential tool for diagnosis of dry mouth including Sjögren's syndrome, Oral Diseases, 10.1111/odi.12252, 21, 2, 224-231, 2015.03, Objectives: Recently, the use of saliva as a diagnostic tool has gained considerable attention because it is non-invasive and easy to perform repeatedly. In this study, we focused on soluble molecules in saliva to establish a new diagnostic method for xerostomia. Materials and Methods: Saliva was obtained from 90 patients with Sjögren's syndrome (SS), 22 patients with xerostomia associated with neurogenic/neuropsychiatric disorders and drugs (XND), 30 patients with radiation-induced xerostomia (RX), and 36 healthy controls. Concentrations of helper T (Th) cytokines in saliva were measured by flow cytometric analysis. Concentrations of secretory IgA (SIgA) and chromogranin A (CgA) were measured by ELISA. Results: Unstimulated and stimulated whole saliva from patients with SS, XND, and RX was significantly reduced compared with controls. Th1 and Th2 cytokines from SS patients were significantly higher than controls. Furthermore, Th2 cytokines were closely associated with strong lymphocytic accumulation in salivary glands from SS patients, while Th1 and Th17 cytokines were negatively associated. SIgA levels were not significantly different between all patient groups and controls. CgA levels from XND patients were significantly higher than controls. Conclusions: The measurement of cytokines, CgA, and SIgA in saliva is suggested to be useful for the diagnosis of xerostomia and also to reveal disease status..
41. Miho Ohta, Masafumi Moriyama, Yuichi Goto, Shintaro Kawano, Akihiko Tanaka, Takashi Maehara, Sachiko Furukawa, Jun-Nosuke Hayashida, Tamotsu Kiyoshima, Mayumi Shimizu, Yojiro Arinobu, Seiji Nakamura, A case of marginal zone B cell lymphoma mimicking IgG4-related dacryoadenitis and sialoadenitis, WORLD JOURNAL OF SURGICAL ONCOLOGY, 10.1186/s12957-015-0459-z, 13, 67, 2015.02, Background: IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS), so-called Mikulicz's disease, is characterized by elevated serum IgG4 and infiltration of IgG4-positive plasma cells in glandular tissues. Recently, several studies reported both malignant lymphoma developed on the background of IgG4-associated conditions and IgG4-producing malignant lymphoma (non-IgG4-related disease).
Case presentation: We report on the case of a 70-year-old man who was strongly suspected IgG4-DS because of high serum IgG4 concentration (215 mg/dl) and bilateral swelling of parotid and submandibular glands. Biopsies of cervical lymph node and a portion of submandibular gland were performed. These histopathological findings subsequently confirmed a diagnosis of marginal zone B cell lymphoma.
Conclusion: Differential diagnosis of IgG4-DS is necessary from other disorders, including Sjogren's syndrome, sarcoidosis, Castleman's disease, Wegener's granulomatosis, lymphoma, and cancer. We suggest that biopsy of swollen lesions is important for a definitive diagnosis of IgG4-DS and discuss the mechanism of development in this case..
42. Jun-Nosuke Hayashida, Sakae Minami, Masafumi Moriyama, Takeshi Toyoshima, Shouichi Shinozaki, Akihiko Tanaka, Takashi Maehara, Seiji Nakamura, Differences of stimulated and unstimulated salivary flow rates in patients with dry mouth, Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology, 10.1016/j.ajoms.2014.04.011, 27, 1, 96-101, 2015.01, Purpose: The purpose of this study was to clarify the usefulness of noninvasive examination items such as sialometry and Visual Analog Scale (VAS) in distinguishing Sjögren's syndrome (SS) in dry mouth patients from neurogenic/neuropsychiatric disorders and drugs (DND). Patients and methods: The study cohort comprised 50 patients with SS and 28 patients with DND. The gum test and Saxon test for stimulated salivary flow rate (SSFR), the spitting test for unstimulated salivary flow rate (USFR) and VAS were performed in all the patients with dry mouth. Results: In SS patients, the SSFR (mean: gum test, 6.34 mL/10. min
Saxon test, 1.19 g/2. min) and USFR (0.61 mL/15. min) were decreased. In DND patients, the SSFR (gum test, 16.35. mL/10 min
Saxon test, 3.58 g/2 min) was within the normal range, but the USFR (0.90 mL/15 min) was decreased. In VAS, SS patients scored significantly higher in the items of "water-drinking at meals", "difficulty in swallowing", and "taste abnormality", while significantly lower in the item of "oral pain". Conclusion: These results suggest that the SSFR, USFR and VAS could be useful in distinguishing DND from SS..
43. Sachiko Furukawa, Masafumi Moriyama, Akihiko Tanaka, Takashi Maehara, Hiroto Tsuboi, Mana Iizuka, Jun-Nosuke Hayashida, Miho Ohta, Takako Saeki, Kenji Notohara, Takayuki Sumida, Seiji Nakamura, Preferential M2 macrophages contribute to fibrosis in IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease, CLINICAL IMMUNOLOGY, 10.1016/j.clim.2014.10.008, 156, 1, 9-18, 2015.01, IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS) is characterized by bilateral swelling of glandular tissues with extensive fibrosis, and is immunologically considered a Th2-predominant disease. Recent studies reported that alternatively activated (M2) macrophages enhanced Th2 immune responses and fibrosis by production of pro-fibrotic factors (IL-10, IL-13 and CCL18). Therefore, we examined the association between M2 macrophages and fibrosis in submandibular glands from 7 patients with IgG4-DS, 10 patients with chronic sialoadenitis, 10 patients with Sjogren's syndrome, and 10 healthy subjects. The number of M2 macrophages in SMGs from patients with IgG4-DS was also significantly higher than in the other groups. Double immunofluorescence staining showed that IL-10 and CCL18 expression co-localized with M2 macrophage-marker (CD163). Furthermore, the SMG fibrosis score was positively correlated with the frequency of M2 macrophages in only IgG4-DS. These results indicate that IL-10 and CCL18 secreted by preferential M2 macrophages possibly play a key role in the development of severe fibrosis in IgG4-DS. (C) 2014 The Authors. Published by Elsevier Inc..
44. Tsuboi H, Nakai Y, Iizuka M, Asashima H, Hagiya C, Tsuzuki S, Hirota T, Miki H, Hagiwara S, Kondo Y, Tanaka A, Moriyama M, Matsumoto I, Nakamura S, Yoshihara T, Abe K, Sumida T, DNA microarray analysis of labial salivary glands in IgG4-related disease: comparison with Sjögren's syndrome., Arthritis & rheumatology (Hoboken, N.J.), 10.1002/art.38748, 66, 10, 2892-2899, 2014.10.
45. Moriyama M, Tanaka A, Maehara T, Furukawa S, Nakashima H, Nakamura S, T helper subsets in Sjögren's syndrome and IgG4-related dacryoadenitis and sialoadenitis: a critical review., Journal of autoimmunity, 10.1016/j.jaut.2013.07.007, 51, 81-88, 2014.06.
46. Shinsuke Ieda, Masafumi Moriyama, Toru Takashita, Takashi Maehara, Yumi Imabayashi, Shoichi Shinozaki, Akihiko Tanaka, Jun-Nosuke Hayashida, Sachiko Furukawa, Miho Ohta, Yoshihisa Yamashita, Seiji Nakamura, Molecular Analysis of Fungal Populations in Patients with Oral Candidiasis Using Internal Transcribed Spacer Region, PLOS ONE, 10.1371/journal.pone.0101156, 9, 6, e101156, 2014.06, Oral candidiasis is closely associated with changes in the oral fungal flora and is caused primarily by Candida albicans. Conventional methods of fungal culture are time-consuming and not always conclusive. However, molecular genetic analysis of internal transcribed spacer (ITS) regions of fungal rRNA is rapid, reproducible and simple to perform. In this study we examined the fungal flora in patients with oral candidiasis and investigated changes in the flora after antifungal treatment using length heterogeneity-polymerization chain reaction (LH-PCR) analysis of ITS regions. Fifty-two patients with pseudomembranous oral candidiasis (POC) and 30 healthy controls were included in the study. Fungal DNA from oral rinse was examined for fungal species diversity by LH-PCR. Fungal populations were quantified by real-time PCR and previously-unidentified signals were confirmed by nucleotide sequencing. Relationships between the oral fungal flora and treatment-resistant factors were also examined. POC patients showed significantly more fungal species and a greater density of fungi than control individuals. Sixteen fungi were newly identified. The fungal populations from both groups were composed predominantly of C. albicans, though the ratio of C. dubliniensis was significantly higher in POC patients than in controls. The diversity and density of fungi were significantly reduced after treatment. Furthermore, fungal diversity and the proportion of C. dubliniensis were positively correlated with treatment duration. These results suggest that C. dubliniensis and high fungal flora diversity might be involved in the pathogenesis of oral candidiasis. We therefore conclude that LH-PCR is a useful technique for diagnosing and assessing the severity of oral candidal infection..
47. Tsuboi H, Asashima H, Takai C, Hagiwara S, Hagiya C, Yokosawa M, Hirota T, Umehara H, Kawakami A, Nakamura H, Sano H, Tsubota K, Ogawa Y, Takamura E, Saito I, Inoue H, Nakamura S, Moriyama M, Takeuchi T, Tanaka Y, Hirata S, Mimori T, Yoshifuji H, Ohta A, Matsumoto I, Sumida T, Primary and secondary surveys on epidemiology of Sjögren's syndrome in Japan., Modern rheumatology, 10.3109/14397595.2013.843765, 24, 3, 464-470, 2014.05.
48. Masafumi Moriyama, Seiji Nakamura, Salivary glands in mikulicz’s disease, Igg4-Related Disease, 10.1007/978-4-431-54228-5_13, 85-92, 2014.01, Mikulicz’s disease (MD) was considered to be a subtype of Sjögren’s syndrome (SS), based on histopathological similarities. However, recent studies have indicated that patients with MD show high serum IgG4 concentrations and suggested that MD is an “IgG4-related disease” and distinguishable from SS. We examined the clinical and serological features of MD and SS in detail to determine the ways in which the two conditions can be differentiated. MD can be diagnosed quickly and simply using diagnostic criteria such as serum IgG4 values and the presence of IgG4+ cells in the salivary glands, in conjunction with salivary gland imaging, sonography, and CT scans. Glucocorticoid therapy of MD reduces IgG and IgG4 levels and improves salivary function. A negative correlation between disease duration and increasing rate of salivary flow was observed in MD..
49. Masafumi Moriyama, S. Furukawa, Shintarou Kawano, Y. Goto, Tamotsu Kiyoshima, A. Tanaka, T. Maehara, Hayashida Jun-Nosuke, M. Ohta, Seiji Nakamura, The diagnostic utility of biopsies from the submandibular and labial salivary glands in IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease, International Journal of Oral and Maxillofacial Surgery, 10.1016/j.ijom.2014.06.014, 43, 10, 1276-1281, 2014.01, IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS) is characterized by serum IgG4 elevation and the infiltration of IgG4-positive plasma cells in glandular tissues. For definitive diagnosis of IgG4-DS, biopsies of local lesions are recommended to exclude Sjögren's syndrome (SS), malignant tumours, and similar disorders. In this study, we examined the diagnostic utility of submandibular gland (SMG) and labial salivary gland (LSG) biopsies in IgG4-DS. Fourteen patients presenting with swelling of the SMG (eight females and six males) underwent both SMG and LSG biopsies. The sensitivity, specificity, and accuracy of SMG biopsies were all 100.0%. In contrast, those of LSG biopsies were 69.2%, 100.0%, and 71.4%, respectively. Thirty-three out of 61 LSG biopsies (54.1%) from all 14 patients were positive for the diagnostic criteria of IgG4-DS (IgG4-positive/IgG-positive plasma cells >0.4). None of the patients experienced complications such as facial nerve palsy, sialocele, or hyposalivation. The IgG4/IgG ratio showed no significant correlation between the LSG and SMG. The final diagnosis was IgG4-DS in 13 patients and marginal zone B-cell lymphoma (MZL) in one. These results suggest that incisional biopsy of the SMG is useful and appropriate for the definitive diagnosis of IgG4-DS, while diagnosis by LSG biopsy alone requires more caution..
50. Masafumi Moriyama, Seiji Nakamura, Salivary glands in mikulicz’s disease, Igg4-Related Disease, 10.1007/978-4-431-54228-5_13, 85-92, 2014.01, Mikulicz’s disease (MD) was considered to be a subtype of Sjögren’s syndrome (SS), based on histopathological similarities. However, recent studies have indicated that patients with MD show high serum IgG4 concentrations and suggested that MD is an “IgG4-related disease” and distinguishable from SS. We examined the clinical and serological features of MD and SS in detail to determine the ways in which the two conditions can be differentiated. MD can be diagnosed quickly and simply using diagnostic criteria such as serum IgG4 values and the presence of IgG4+ cells in the salivary glands, in conjunction with salivary gland imaging, sonography, and CT scans. Glucocorticoid therapy of MD reduces IgG and IgG4 levels and improves salivary function. A negative correlation between disease duration and increasing rate of salivary flow was observed in MD..
51. Masafumi Moriyama, Seiji Nakamura, New understanding of Mikulicz's disease/IgG4-related disease and Sjögren's syndrome, Japanese Journal of Clinical Radiology, 59, 8, 1033-1043, 2014, IgG4-related disease (IgG4-RD) is a systemic disease characterized by the elevation of serum IgG4 and infiltration of IgG4-positive plasma cells in the target organs, and its molecular mechanism of pathogenesis remains to be clarified. IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS), so-called Mikulicz's disease or Küttner's tumor, is included in IgG4-RD and shows clinical and histological similarities to Sjögren's syndrome (SS). In this review, we focus on the selective localization and respective functions of Th cell subsets and discuss the differences between IgG4-DS and SS to clarify the clinical and pathogenic mechanisms of these diseases..
52. Masafumi Moriyama, Akihiko Tanaka, Takashi Maehara, Yukiko Ohyama, Mayumi Shimizu, Hitoshi Nakashima, Jun-Nosuke Hayashida, Shoichi Shinozaki, Yoshiaki Kubo, Sachiko Furukawa, Toshihiro Kikuta, Seiji Nakamura, Clinical characteristics of Mikulicz's disease as an IgG4-related disease, CLINICAL ORAL INVESTIGATIONS, 10.1007/s00784-012-0905-z, 17, 9, 1995-2002, 2013.12, Mikulicz's disease (MD) was considered to be a subtype of Sjogren's syndrome (SS), based on histopathological similarities. However, recent studies have indicated that patients with MD show high serum IgG4 concentration, and suggested that MD is one of "IgG4-related disease" and distinguishable from SS. Therefore, we clinically and histopathologically examined the disease states of MD and SS in detail.
Twenty patients with Mikulicz's disease and 18 with SS were comparatively studied to determine clinical characteristics in MD patients.
Sialography in MD patients did not show the "apple-tree sign" typically seen in SS. Serologically, high serum IgG4 levels but not anti-SS-A or anti-SS-B antibodies were observed in MD. SS showed lymphocytic infiltration of various subsets with atrophy or severe destruction of the acini, while MD showed selective infiltration of IgG4+ plasma cells with hyperplastic germinal centers and mild acini destruction. Corticosteroid treatment of MD reduced IgG and IgG4 levels and improved salivary function. A negative correlation between disease duration and increasing rate of salivary flow was observed in MD.
These results suggested that the pathogenesis of MD might be different from those of SS. Clinical Relevance: early diagnosis and treatment of MD is important for the improvement of salivary function..
53. Tsuboi,H, Nakai,Y, Iizuka,M, Asashima,H, Kondo,Y, Tanaka,A, Moriyama,M, Matsumoto,I, Yoshihara,T, Nakamura,S, Abe,K, Sumida,T, ANALYSIS OF MOLECULAR MECHANISM IN IGG4-RELATED DISEASE: COMPARISON WITH SJOGREN'S SYNDROME, ANNALS OF THE RHEUMATIC DISEASES, 72, 3, 466-466, 2013.06.
54. Tsuboi H, Hagiwara S, Asashima H, Umehara H, Kawakami A, Nakamura H, Sano H, Tsubota K, Ogawa Y, Takamura E, Saito I, Inoue H, Nakamura S, Moriyama M, Takeuchi T, Tanaka Y, Hirata S, Mimori T, Matsumoto I, Sumida T, Validation of different sets of criteria for the diagnosis of Sjögren's syndrome in Japanese patients., Modern rheumatology, 10.1007/s10165-012-0812-9, 23, 2, 219-225, 2013.03.
55. Hayashida Jun-Nosuke, Seiji Nakamura, T. Toyoshima, Masafumi Moriyama, M. Sasaki, E. Kawamura, Yukiko Ohyama, Kumamaru Wataru, K. Shirasuna, Possible involvement of cytokines, chemokines and chemokine receptors in the initiation and progression of chronic GVHD, Bone Marrow Transplantation, 10.1038/bmt.2012.100, 48, 1, 115-123, 2013.01, Chronic GVHD (cGVHD) after allogeneic hematopoietic SCT (HSCT) is characterized by an infiltration of T cells into target organs including the oral mucosa and salivary glands. This study was designed to clarify the molecular mechanism of the local accumulation of pathogenic T cells in cGVHD. The expression of cytokines, chemokines and chemokine receptors in the buccal mucosa (BM), labial salivary glands (LSG) and PBMC from 16 patients with cGVHD after allogeneic HSCT was examined. The mRNA expression of T helper 1 (Th1) and Th2 cytokines, and several chemokines and chemokine receptors was significantly increased in the BM and LSG from cGVHD patients, in comparison with both those in the BM and LSG from controls, respectively, and also with those in the PBMC from cGVHD patients. Furthermore, the mRNA expression of Th2 cytokines, macrophage-derived chemokine and CC chemokine receptor 4 was closely associated with a strong T-cell infiltration in the BM and LSG from cGVHD patients. These results suggest that cGVHD might be initiated and/or maintained by Th1/Th0 cells and thereafter progresses in association with Th2 cell accumulation via the interaction of particular chemokine and chemokine receptors..
56. Takashi Maehara, Masafumi Moriyama, Hitoshi Nakashima, Katsuhisa Miyake, Jun-Nosuke Hayashida, Akihiko Tanaka, Shouichi Shinozaki, Yoshiaki Kubo, Seiji Nakamura, Interleukin-21 contributes to germinal centre formation and immunoglobulin G4 production in IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease, ANNALS OF THE RHEUMATIC DISEASES, 10.1136/annrheumdis-2012-201477, 71, 12, 2011-2019, 2012.12, Objectives Interleukin (IL)-21 is mainly produced by CD4 T helper (Th) cells including Th2, Th17 and follicular helper T (Tfh) cells. Recent studies have reported that IL-21 is involved in the formation of germinal centres (GCs) and class switching of IgG4. It has been suggested that IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS), so-called Mikulicz's disease (MD), is distinct from Sjogren's syndrome (SS) and shows a high frequency of GC formation in salivary glands. In this study the expression of IL-21 in IgG4-DS and SS patients was examined.
Methods Twelve patients with IgG4-DS, 15 with SS and 15 healthy subjects were screened for (1) ectopic GC formation in formalin-fixed labial salivary gland (LSG) biopsy samples; (2) expression of IL-21, Th2-, Th17- and Tfh-related molecules (cytokines, chemokine receptors and transcription factors) in LSGs; (3) relationship between IgG4/IgG ratio and mRNA expression of IL-21 in LSGs.
Results mRNA expression of IL-21 and Bcl-6 in LSGs from patients with IgG4-DS was significantly higher than in patients with SS and controls. IL-21 and CXCR5 were detected by immunohistochemistry in or around GC in patients with SS and those with IgG4-DS. IL-21 was detected in infiltrating lymphocytes outside GC only in patients with IgG4-DS. Expression of IL-21 was consistent with that of Th2-related molecules while IL-17 was rarely seen in IgG4-DS. Furthermore, the expression of IL-21 in LSGs was correlated with the number of GC formations and the IgG4/IgG ratio in patients with IgG4-DS.
Conclusions These results suggest that overexpression of IL-21 by Th2 cells might play a key role in GC formation and IgG4 production in IgG4-DS..
57. S. Shinozaki, Masafumi Moriyama, Hayashida Jun-Nosuke, A. Tanaka, T. Maehara, S. Ieda, Seiji Nakamura, Close association between oral Candida species and oral mucosal disorders in patients with xerostomia, Oral Diseases, 10.1111/j.1601-0825.2012.01923.x, 18, 7, 667-672, 2012.10, Objective: Heightened interest in oral health has lead to an increase in patients complaining of xerostomia, which is associated with various oral mucosal disorders. In this study, we investigated the relationship between Candida species and oral mucosal disorders in patients with xerostomia. Subjects and Methods: We evaluated whole salivary flow rate and presence of oral mucosal disorders in 48 patients with xerostomia and 15 healthy controls. The number of Candida species was measured as colony-forming units after propagation on selective medium. Identification of Candida at the species level was carried out by polymerase chain reaction and restriction fragment length polymorphism analysis. We then examined the relationship between Candida species and oral mucosal symptoms. Results: Compared with controls, patients with xerostomia exhibited significantly decreased whole salivary flow rate, increased rate of oral mucosal symptoms, and higher numbers of Candida. Salivary flow rate negatively correlated with the number Candida. Among patients with oral candidiasis, Candida albicanswas isolated from the tongue mucosa and Candida glabratawas isolated from the angle of the mouth. Conclusion: These results suggest that particular Candida species are involved in the pathogenesis of oral mucosal disorders in patients with xerostomia..
58. Molecular Mechanism of IgG4 Class Switch Recombination in IgG4-Related Disease..
59. In vitro induction of specific CD8+ T lymphocytes by tumor-associated antigenic peptides in patients with oral squamous cell carcinoma..
60. T. Maehara, Masafumi Moriyama, Hayashida Jun-Nosuke, A. Tanaka, S. Shinozaki, Y. Kubo, Kaori Matsumura, Seiji Nakamura, Selective localization of T helper subsets in labial salivary glands from primary Sjögren's syndrome patients, Clinical and Experimental Immunology, 10.1111/j.1365-2249.2012.04606.x, 169, 2, 89-99, 2012.08, The aim of this study was to investigate the initiation and progression of autoimmune damage in the lesions of labial salivary glands (LSGs) from primary Sjögren's syndrome (SS) patients by examining the selective localization of T helper (Th) subsets such as Th1, Th2, Th17 regulatory T cells (T regs) and follicular T helper cells (Tfh). The expression of cytokines and transcription factors associated with these Th subsets in the LSGs from 54 SS patients and 16 healthy controls was examined using real-time polymerase chain reaction (PCR) and immunostaining. Additionally, infiltrating lymphocytes without germinal centre (GC -) and with GC (GC +) in the LSGs specimens from eight SS patients were extracted selectively by laser capture microdissection (LCM). The mRNA expression of these molecules was compared between the two sample groups of GC - and GC + by real-time PCR. The mRNA expression of cytokines and transcription factors of all T helper (Th) subsets in the LSGs from the SS patients was increased significantly in comparison with controls. In LSGs from the SS patients, Th2 and Tfh was associated closely with strong lymphocytic infiltration; however, Th1, Th17 and T regs was not. In the selectively extracted lesions of LSGs, Th1 and Th17-related molecules were detected strongly in the GC -, while Th2 and Tfh-related molecules were detected in the GC +. In contrast, no significant association with strong lymphocytic infiltration was observed in T reg-related molecules. These results indicate that SS has selective localization of Th subsets such as Th1, Th2, Th17 and Tfh in the LSGs, which is associated closely with disease severity and/or status. SS might be initiated by Th1 and Th17 cells, and then progressed by Th2 and Tfh cells via GC formation..
61. Hiroto Tsuboi, Naomi Matsuo, Mana Iizuka, Sayaka Tsuzuki, Yuya Kondo, Akihiko Tanaka, Masafumi Moriyama, Isao Matsumoto, Seiji Nakamura, Takayuki Sumida, Analysis of IgG4 class switch-related molecules in IgG4-related disease, Arthritis Research and Therapy, 10.1186/ar3924, 14, 4, 2012.07, Introduction: Immunoglobulin G4 (IgG4)-related disease (IgG4-RD) is a new disease entity characterized by high serum IgG4 levels, IgG4-positive plasmacytic infiltration, and fibrosis in various organs. The purpose of this study was to determine the mechanism of upregulation of IgG4 class switch recombination in IgG4-RD.Methods: We extracted RNA from peripheral blood mononuclear cells (PBMCs) of patients with IgG4-RD (n = 6), Sjögren syndrome (SS) (n = 6), and healthy controls (n = 8), from CD3-positive T cells and CD20-positive B cells sorted from PBMCs of patients with IgG4-RD (n = 3), SS (n = 4), and healthy controls (n = 4), as well as from labial salivary glands (LSGs) of patients with IgG4-RD (n = 11), SS (n = 13), and healthy controls (n = 3). The mRNA expression levels of IgG4-specific class switch-related molecules, such as Th2 cytokines (IL-4 and IL-13), Treg cytokines (IL-10 and TGF-β), and transcriptional factors (GATA3 and Foxp3) were examined with quantitative polymerase chain reaction (PCR). IgG4-nonspecific class switch-related molecules, such as CD40, CD154, BAFF, APRIL, IRF4, and AID, were also examined.Results: The expression levels of Treg cytokines (IL-10 and TGF-β) and AID were significantly higher in LSGs of IgG4-RD than in SS and the controls (P
62. Masafumi Moriyama, Hayashida Jun-Nosuke, T. Toyoshima, Yukiko Ohyama, S. Shinozaki, A. Tanaka, T. Maehara, Seiji Nakamura, Cytokine/chemokine profiles contribute to understanding the pathogenesis and diagnosis of primary Sjögren's syndrome, Clinical and Experimental Immunology, 10.1111/j.1365-2249.2012.04587.x, 169, 1, 17-26, 2012.07, To investigate the pathogenesis of localized autoimmune damage in Sjögren's syndrome (SS) by examining the expression patterns of cytokines, chemokines and chemokine receptors at sites of autoimmune damage. mRNA expression of these molecules in the labial salivary glands (LSGs) and peripheral blood mononuclear cells (PBMCs) from 36 SS patients was examined using a real-time polymerase chain reaction-based method. Subsets of the infiltrating lymphocytes and chemokines/chemokine receptors expression in the LSG specimens were examined by immunohistochemistry. Cytokines/chemokine concentrations in the saliva were analysed using flow cytometry or enzyme-linked immunosorbent assay. mRNA expression of T helper type 1 (Th1) cytokines, chemokines and chemokine receptors was higher in LSGs than in PBMCs. In contrast, mRNA expression of Th2 cytokines, chemokines [thymus and activation-regulated chemokine (TARC/CCL17), macrophage-derived chemokine (MDC/CCL22)] and chemokine receptor (CCR4) was associated closely with strong lymphocytic accumulation in LSGs. Furthermore, TARC and MDC were detected immunohistochemically in/around the ductal epithelial cells in LSGs, whereas CCR4 was detected on infiltrating lymphocytes. The concentrations of these cytokines/chemokines were significantly higher in the saliva from SS patients than those from controls, and the concentrations of Th2 cytokines/chemokines were associated closely with strong lymphocytic accumulation in LSGs. These results suggest that SS might be initiated and/or maintained by Th1 and Th17 cells and progress in association with Th2 cells via the interaction between particular chemokines/chemokine receptors. Furthermore, the measurement of cytokines/chemokines in saliva is suggested to be useful for diagnosis and also to reveal disease status..
63. Akihiko Tanaka, Masafumi Moriyama, Hitoshi Nakashima, Katsuhisa Miyake, Jun-Nosuke Hayashida, Takashi Maehara, Shouichi Shinozaki, Yoshiaki Kubo, Seiji Nakamura, Th2 and regulatory immune reactions contribute to IgG4 production and the initiation of Mikulicz disease, ARTHRITIS AND RHEUMATISM, 10.1002/art.33320, 64, 1, 254-263, 2012.01, Objective Mikulicz disease has been considered to be a subtype of Sjogren's syndrome (SS). However, recent studies have suggested that Mikulicz disease is an IgG4-related disease and is distinguishable from SS. In addition, it has been reported that both interleukin-4 (IL-4) and IL-10 induce IgG4 production and inhibit IgE. This study was undertaken to examine the expression of these cytokines in patients with Mikulicz disease and patients with SS.
Methods. Labial salivary gland (LSG) sections from 15 patients with Mikulicz disease and 18 patients with SS were examined for subsets of the infiltrating lymphocytes, expression patterns of messenger RNA (mRNA) for cytokines/chemokines, and relationships between the IgG4:IgG ratio and the expression of mRNA for IL-4 or IL-10.
Results. Immunohistochemical analysis showed lymphocyte infiltration of various subsets in the LSGs of SS patients, and the selective infiltration of IgG4-positive plasma cells and Treg cells in the LSGs of Mikulicz disease patients. The levels of mRNA for both Th1 and Th2 cytokines and chemokines in LSGs from patients with SS were significantly higher than in controls, while the expression of both Th2 and Treg cells was significantly higher in the patients with Mikulicz disease than in controls. Furthermore, the expression of IL-4 or IL-10 in the LSGs was correlated with the IgG4: IgG ratio.
Conclusion. These results suggest that the pathogenesis of Mikulicz disease is different from that of SS. Mikulicz disease is a unique inflammatory disorder characterized by Th2 and regulatory immune reactions that might play key roles in IgG4 production..
64. H. Nakashima, K. Miyake, Masafumi Moriyama, A. Tanaka, M. Watanabe, Y. Abe, H. Sato, Seiji Nakamura, T. Saito, An amplification of IL-10 and TGF-β in patients with IgG4-related tubulointerstitial nephritis, Clinical Nephrology, 73, 5, 385-391, 2010.05, Background: IgG4-related tubulointerstitial nephritis (TIN) shows characteristic serum IgG4 elevation and increased IgG4-positive plasma cells in the renal interstitium, and inclusion of TIN as an IgG4-related systemic disease has been suggested. IgG4 is the rarest IgG subclass and is a Th2-dependent isotype with low affinity for target antigen. Although the pathogenesis of this disease has not been elucidated, positive serum immune complex and hypocomplementemia in some patients with this disease suggest that immune complex mechanisms are involved in the causation of this disease. Method:We selected 20 cases of histological diagnosed TIN. These cases were etiologically different and included 4 cases of IgG4-related TIN.We extracted RNAfrom paraffin embedded biopsied kidney and evaluated expression levels of various cytokines for each case by real time PCR. Results: Comparison of cytokine production patterns among different disease-associated TINs revealed that IgG4-related TIN exhibited a quite distinct pattern. On the one hand, there was no expression of IL-2, IFN-γ, IL-17 and IL-6, whereas production of IL-4, IL-10 and TGF-β was, on the other hand, remarkably increased in IgG4-related TIN. Conclusion: Based on these cytokine production results, Th2 and Treg appear to play a central role in IgG4-related TIN..
65. Mayumi Shimizu, Masafumi Moriyama, Kazutoshi Okamura, Toshiyuki Kawazu, Toru Chikui, Tazuko K. Goto, Yukiko Ohyama, Seiji Nakamura, Kazunori Yoshiura, Sonographic diagnosis for Mikulicz disease, ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTOLOGY, 10.1016/j.tripleo.2009.02.032, 108, 1, 105-113, 2009.07, Objective. The aim was to investigate the diagnostic imaging characteristics of Mikulicz disease (MD), especially sonographic ones, and to clarify the differences between them and those in Sjogren syndrome (SS), based on new criteria of MD.
Study design. The sonographic and sialographic images, as well as clinical, histopathologic, and serologic findings of 9 patients satisfying the new criteria of MD were analyzed and compared with those in SS.
Results. All swollen submandibular glands showed bilateral nodal hypoechoic areas with high vascularization on sonograms and a parenchymal defect on sialograms, whereas parotid glands showed normal or slight change on both images. Nodal areas in submandibular gland sonograms were unclear on computerized tomography and on magnetic resonance imaging, but showed accumulation on gallium scintigraphy.
Conclusion. Mikulicz disease showed a high rate of bilateral nodal change in submandibular glands, which was completely different from SS. For detection and follow-up of these changes, sonography may be the best imaging modality. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108: 105-113).
66. Katsuhisa Miyake, Masafumi Moriyama, Kumiko Aizawa, Shuji Nagano, Yasushi Inoue, Atsushi Sadanaga, Hitoshi Nakashima, Seiji Nakamura, Peripheral CD4+T cells showing a Th2 phenotype in a patient with Mikulicz's disease associated with lymphadenopathy and pleural effusion, MODERN RHEUMATOLOGY, 10.1007/s10165-007-0010-3, 18, 1, 86-90, 2008.02, Mikulicz's disease (MD) is a unique IgG4-related systemic disease indicated by enlargement of the lachrymal and salivary glands and which differs substantially from Sjogren's syndrome. A male patient with pleural effusion, swelling of the submandibular glands, and swelling of the paraaortic, mediastinal, and pararenal lymph nodes was diagnosed with MD. Analysis of peripheral CD4+ T cells from the patient revealed deviation of the Th1/Th2 balance to Th2. Prednisolone therapy ameliorated the disease and corrected the Th1/Th2 imbalance..
67. T. Toyoshima, Seiji Nakamura, Kumamaru Wataru, E. Kawamura, H. Ishibashi, Hayashida Jun-Nosuke, Masafumi Moriyama, Yukiko Ohyama, M. Sasaki, K. Shirasuna, Expression of tumor-associated antigen RCAS1 and its possible involvement in immune evasion in oral squamous cell carcinoma, Journal of Oral Pathology and Medicine, 10.1111/j.1600-0714.2006.00442.x, 35, 6, 361-368, 2006.07, BACKGROUND: RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is known to induce apoptosis in its receptor-positive cells. The authors investigated RCAS1 expression in oral squamous cell carcinoma (SCC) and its association with the apoptosis of tumor-infiltrating lymphocytes (TILs). METHODS: In 130 patients with oral SCC, the expression of RCAS1 in tumor cells was immunohistochemically examined and the apoptosis of TILs was examined by Terminal Deoxynucleotidyltransferase-mediated dUTP Nick End Labeling (TUNEL) staining. RESULTS: RCAS1 was detected both on the cytoplasm and the membrane of tumor cells in 41 of 130 cases (31.5%). Focusing on the expression at the invasive front interacting with host immune cells, RCAS1 was detected in 22 of 130 cases (16.9%). The percentage of TUNEL-positive TILs in cases with RCAS1-positive SCCs was significantly higher than in cases with RCAS1-negative SCCs (P