Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Yojiro Arinobu Last modified date:2021.06.18

Lecturer / Department of Clinical Immunology and Rheumatology / Infectious Disease / Kyushu University Hospital

1. Yojiro Arinobu , Yusuke Kashiwado, Kohta Miyawaki, Masahiro Ayano, Yasutaka Kimoto, Hiroki Mitoma, Mitsuteru Akahoshi, Toshihiro Miyamoto, Takahiko Horiuchi, Koichi Akashi, Hiroaki Niiro, Autoimmune manifestations associated with myelodysplastic syndrome predict a poor prognosis., Medicine, 100, 13, e25406, 2021.04.
2. K. Higashioka, Y. Kikushige, M. Ayano, Y. Kimoto, H. Mitoma, M. Kikukawa, M. Akahoshi, Y. Arinobu, T. Horiuchi, K. Akashi, H. Niiro, Generation of a novel CD30+ B cell subset producing GM-CSF and its possible link to the pathogenesis of systemic sclerosis, Clinical and Experimental Immunology, 10.1111/cei.13477, 201, 3, 233-243, 2020.09, Systemic sclerosis (SSc) is a T helper type 2 (Th2)-associated autoimmune disease characterized by vasculopathy and fibrosis. Efficacy of B cell depletion therapy underscores antibody-independent functions of B cells in SSc. A recent study showed that the Th2 cytokine interleukin (IL)-4 induces granulocyte–macrophage colony-stimulating factor (GM-CSF)-producing effector B cells (GM-Beffs) in humans. In this study, we sought to elucidate the generation mechanism of GM-Beffs and also determine a role of this subset in SSc. Among Th-associated cytokines, IL-4 most significantly facilitated the generation of GM-Beffs within memory B cells in healthy controls (HCs). In addition, the profibrotic cytokine transforming growth factor (TGF)-β further potentiated IL-4- and IL-13-induced GM-Beffs. Of note, tofacitinib, a Janus kinase (JAK) inhibitor, inhibited the expression of GM-CSF mRNA and protein in memory B cells induced by IL-4, but not by TGF-β. GM-Beffs were enriched within CD20+CD30+CD38−/low cells, a distinct population from plasmablasts, suggesting that GM-Beffs exert antibody-independent functions. GM-Beffs were also enriched in a CD30+ fraction of freshly isolated B cells. GM-Beffs generated under Th2 conditions facilitated the differentiation from CD14+ monocytes to DC-SIGN+CD1a+CD14CD86+ cells, which significantly promoted the proliferation of naive T cells. CD30+ GM-Beffs were more pronounced in patients with SSc than in HCs. A subpopulation of SSc patients with the diffuse type and concomitant interstitial lung disease exhibited high numbers of GM-Beffs. Together, these findings suggest that human GM-Beffs are enriched in a CD30+ B cell subset and play a role in the pathogenesis of SSc..
3. Qiaolei Wang, Daisuke Oryoji, Hiroki Mitoma, Yasutaka Kimoto, Masamichi Koyanagi, Kana Yokoyama, Masahiro Ayano, Mitsuteru Akahoshi, Yojiro Arinobu, Hiroaki Niiro, Koichi Akashi, Takahiko Horiuchi, Methotrexate Enhances Apoptosis of Transmembrane TNF-Expressing Cells Treated With Anti-TNF Agents, Frontiers in Immunology, 10.3389/fimmu.2020.02042, 11, 2020.08, Background: Concomitant use of methotrexate (MTX) improves the clinical efficacy of anti-TNF agents in the treatment of rheumatoid arthritis (RA). We aimed to clarify the cytotoxic effect of MTX on transmembrane TNF (tmTNF)-expressing cells treated with anti-TNF agents. Methods: Jurkat T cells stably expressing tmTNF were used for the following experiments. Cytotoxicity induced by an anti-TNF agent (infliximab, adalimumab, or certolizumab pegol) with concomitant MTX were compared with that by MTX alone or by an anti-TNF agent alone using flow cytometry. Apoptosis-induction mediated by reverse signal through tmTNF, complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP) were evaluated. Folic acid and Y-27632, a Rho kinase inhibitor, were used as inhibitors to study intracellular signaling pathway in apoptosis induced by MTX and anti-TNF agents. Results: Apoptosis of tmTNF-expressing cells was significantly increased by the concomitant administration of MTX and an anti-TNF agent, compared with MTX alone or an anti-TNF agent alone. The apoptosis induction by concomitant MTX was most pronounced in infliximab-treatment. Reverse signal transduction, but not CDC or ADCC/ADCP, was responsible for the coordinate effect of MTX and an anti-TNF agent on tmTNF-expressing cells. Folic acid inhibited MTX-mediated apoptosis, while Y-27632 suppressed JNK activation and infliximab-induced apoptosis via revere signal through tmTNF. Conclusion: The apoptotic effect was enhanced by combination of MTX and an anti-TNF agent in tmTNF-expressing cells. The intracellular pathways induced by MTX and anti-TNF agents seem to be independent. These findings might explain at least in part improved the clinical response upon co-therapy of MTX and an anti-TNF agent in RA..
4. Yushiro Endo, Shin ya Kawashiri, Shimpei Morimoto, Ayako Nishino, Momoko Okamoto, Sosuke Tsuji, Ayuko Takatani, Toshimasa Shimizu, Remi Sumiyoshi, Takashi Igawa, Tomohiro Koga, Naoki Iwamoto, Kunihiro Ichinose, Mami Tamai, Hideki Nakamura, Tomoki Origuchi, Yukitaka Ueki, Tamami Yoshitama, Nobutaka Eiraku, Naoki Matsuoka, Akitomo Okada, Keita Fujikawa, Hiroaki Hamada, Tomomi Tsuru, Shuji Nagano, Yojiro Arinobu, Toshihiko Hidaka, Yoshifumi Tada, Atsushi Kawakami, Non-TNF inhibitor switchers versus TNF inhibitor cyclers from multicentre rheumatoid arthritis ultrasonography prospective cohort in Japan, Immunological Medicine, 10.1080/25785826.2020.1757920, 43, 3, 115-120, 2020.07, To compare therapeutic efficacy of tumour necrosis factor inhibitor (TNFi) cyclers and non-TNFi switchers in patients with rheumatoid arthritis (RA) having inadequate response to previous TNFis (TNF-IR patients) using composite measures including imaging assessment with power Doppler ultrasonography (PDUS). Patients with RA who had inadequate response to one or more previous TNFi agents with moderate or higher disease activity were enrolled. The outcomes of 56 TNF-IR patients were analysed. Patients were divided into 19 TNFi cyclers and 37 non-TNFi switchers (16 abatacept [ABT] and 21 tocilizumab [TCZ] switchers). Retention ratio at 6 months was significantly higher in non-TNFi switchers than in TNFi cyclers (p '.05). Although there was no significant difference, non-TNFi switchers tended to have a larger decrease than TNFi cyclers in efficacy indicators based on clinical disease activity index and PDUS. Multivariate logistic regression analysis identified a following independent factor associated with both EULAR good response and retention of a biologic agent: non-TNFi switch (p '.05 for both). Non-TNFi switchers were shown to have significantly higher percentage of EULAR good response and higher retention than TNFi cyclers. A non-TNFi biologic agent may hence be a preferential next-line treatment for TNF-IR patients..
5. Masahiro Ayano, Yojiro Arinobu, Hiroshi Tsukamoto, Shun ichiro Ota, Kenta Misaki, Keisuke Nishimura, Yasutaka Kimoto, Hiroki Mitoma, Mitsuteru Akahoshi, Koichi Akashi, Takahiko Horiuchi, Hiroaki Niiro, Shoulder ultrasound and serum lactate dehydrogenase predict inadequate response to glucocorticoid treatment in patients with polymyalgia rheumatica, Rheumatology International, 10.1007/s00296-020-04512-9, 40, 7, 1101-1109, 2020.07.
6. Kazuhiko Higashioka, Yuri Ota, Takashi Maehara, Masafumi Moriyama, Masahiro Ayano, Hiroki Mitoma, Mitsuteru Akahoshi, Yojiro Arinobu, Takahiko Horiuchi, Seiji Nakamura, Koichi Akashi, Hiroaki Niiro, Association of circulating SLAMF7+Tfh1 cells with IgG4 levels in patients with IgG4-related disease, BMC Immunology, 10.1186/s12865-020-00361-0, 21, 1, 2020.06, Background: Follicular helper CD4+ T (Tfh) cells have a critical role in IgG4 production by B cells in IgG4-related disease (IgG4-RD). Recent studies including ours showed that SLAMF7+CD4+ T cells are an important pathological driver of IgG4-RD. In this study, we have sought to elucidate a relationship between helper CD4+ T (Th), particularly Tfh, cells and SLAMF7+ CD4+ T cells in IgG4-RD. Results: The patients with IgG4-RD enrolled in this study were aged 66 ± 12 years and their titers of serum IgG4 were 372 ± 336 mg/dl. Th1 cells, activated circulating Tfh1 (cTfh1), and activated cTfh2 cells increased in IgG4-RD. SLAMF7 was mainly expressed on Th1 and cTfh1, but not cTfh2, cells in the patients. SLAMF7+ cTfh1 cells were PD-1/CD28 double-positive, whereas SLAMF7+ Th1 cells were CD28 negative. Positive correlations were noted between serum IgG4 levels and the number of activated cTfh2 cells and SLAMF7+ cTfh1 cells, but not SLAMF7+ Th1 cells. Intriguingly, among cTfh1 cells, activated SLAMF7+ cTfh1 cells were high producers of IL-10 along with IL-21. Blimp-1, but not Bcl-6, mRNA was expressed at high levels in activated SLAMF7+ cTfh1 cells. In addition to CD4+ T cells, the frequency of SLAMF7+ fraction was higher in memory B cells than naïve B cells in patients with IgG4RD. Finally, upon stimulation via B-cell receptor and CD40, Tfh1-associated cytokines, IL-21 and IFN-γ, most significantly induced SLAMF7 expression in memory B cells. Conclusions: Together, these results suggest that circulating SLAMF7+ Tfh1 cells, along with Tfh2 cells, play a pathologic role in IgG4 production in IgG4-RD..
7. Shoichiro Inokuchi, Hiroki Mitoma, Shotaro Kawano, Shota Nakano, Masahiro Ayano, Yasutaka Kimoto, Mitsuteru Akahoshi, Yojiro Arinobu, Hiroshi Tsukamoto, Koichi Akashi, Takahiko Horiuchi, Hiroaki Niiro, Homeostatic milieu induces production of deoxyribonuclease 1-like 3 from myeloid cells, Journal of Immunology, 10.4049/jimmunol.1901304, 204, 8, 2088-2097, 2020.04, DNase 1-like 3 (DNase1L3), which belongs to DNase1 family, was originally identified as one of apoptosis- and necrosis-related endonucleases that fragmentate intranucleosomal DNA. A loss-of-function mutation has been reported in murine models of systemic lupus erythematosus (SLE) and in familial SLE patients. These reports suggest DNase1L3 plays an important role in the prevention of developing SLE; however, expression and function of DNase1L3 in human immune systems have been largely unclarified. As previous reports showed DNase1L3 is expressed in hematopoietic organs, we first analyzed expression levels of DNase1L3 in each subset of human peripheral blood cells by quantitative real-time PCR. Plasmacytoid dendritic cells showed the highest expression levels of DNase1L3 mRNA among peripheral blood cells. IL-4 enhanced DNase1L3 expression in monocytes, monocyte-derived dendritic cells, and monocyte-derived macrophages (MDMs), but not in T cells, B cells, or plasmacytoid dendritic cells. Together with IL-4, all-trans retinoic acid and apoptotic cells efficiently upregulated expression of DNalse1L3 in MDMs. As a result of intracellular signaling analysis, Jak1-IRS2-ERK/PI3K pathway was essential for IL-4-induced DNase1L3 expression. IL-4-treated monocyte-derived dendritic cells and MDMs secreted active DNase1L3 protein that could degrade liposome-DNA complexes, which were resistant to DNase1. Our results indicate DNase1L3 is secreted by innate immune cells and may play a critical role in the tissue homeostasis and on prevention of developing autoimmunity by degrading self-DNA..
8. Y. Endo, T. Koga, S. Y. Kawashiri, S. Morimoto, A. Nishino, M. Okamoto, M. Eguchi, S. Tsuji, A. Takatani, T. Shimizu, R. Sumiyoshi, T. Igawa, N. Iwamoto, K. Ichinose, M. Tamai, H. Nakamura, T. Origuchi, Y. Ueki, T. Yoshitama, N. Eiraku, N. Matsuoka, A. Okada, K. Fujikawa, H. Hamada, T. Tsuru, S. Nagano, Y. Arinobu, T. Hidaka, Y. Tada, A. Kawakami, Anti-citrullinated protein antibody titre as a predictor of abatacept treatment persistence in patients with rheumatoid arthritis
a prospective cohort study in Japan, Scandinavian Journal of Rheumatology, 10.1080/03009742.2019.1627411, 49, 1, 13-17, 2020.01, Objective: Successful rheumatoid arthritis (RA) outcome depends on treatment efficacy in the early stages of the disease and its sustainability. It is thus critical to identify factors predicting treatment persistence with biological agents, such as abatacept. We compared clinical profiles, including early changes in autoantibody titres at 3 months, between patients with RA demonstrating sustained persistence and those discontinuing abatacept treatment. Method: We prospectively enrolled 71 and 78 active RA patients treated with abatacept and tumour necrosis factor inhibitors (TNF-Is), respectively, who had previous disease-modifying anti-rheumatic drug) failure. Clinical characteristics were compared between non-continuation and continuation groups stratified according to abatacept or TNF-I persistence for at least 12 months from treatment initiation. Results: Significantly larger decreases in rheumatoid factor titre and anti-citrullinated protein autoantibody (ACPA) titre were observed in the continuation group of abatacept therapy at 3 months, and early reduction in ACPA titre remained a significant and independent predictor of sustained persistence with abatacept in multivariate analysis. In addition, we obtained the area under the receiver operator characteristics curve of 0.904 from a model including baseline ACPA titre and reduction of ACPA titre at 3 months. Sustained reduction of RA disease activity score at 12 months was significantly and independently associated with reduced ACPA titre at 3 months. Conclusions: Persistence with abatacept and sustained therapeutic response are associated with an early reduction in ACPA titre. Prediction of abatacept continuation and efficacy will facilitate the optimal design of therapy in the early stages of RA..
9. Shoichi Fukui, Ayako Kuwahara-Takaki, Nobuyuki Ono, Shuntaro Sato, Tomohiro Koga, Shin ya Kawashiri, Nozomi Iwanaga, Naoki Iwamoto, Kunihiro Ichinose, Mami Tamai, Hideki Nakamura, Tomoki Origuchi, Kiyoshi Migita, Yojiro Arinobu, Hiroaki Niiro, Yoshifumi Tada, Koichi Akashi, Takahiro Maeda, Atsushi Kawakami, Serum levels of fibroblast growth factor-2 distinguish Takayasu arteritis from giant cell arteritis independent of age at diagnosis, Scientific reports, 10.1038/s41598-018-36825-y, 9, 1, 2019.12, Takayasu arteritis (TAK) and giant cell arteritis (GCA) are two major variants of large vessel vasculitis, and age is a major factor in their differential diagnosis. We sought to determine whether the two diseases exist on the same spectrum. We compared the serum levels of multiple cytokines and chemokines in 25 patients with TAK, 20 patients with GCA, and sex- and age-matched healthy donors for either condition (HD-TAK and HD-GCA). To evaluate the effects of age on the levels of cytokines and chemokines, we performed multiple logistic regression analysis using the least absolute shrinkage and selection operator (LASSO) method. The levels of IL-1RA, IL-10, GM-CSF, G-CSF, FGF-2, eotaxin, and IP-10 were significantly different between TAK and GCA, but no differences were found in the levels of IL-6, IL-12(p40), IL-17, IFN-γ, and TNF-α. Significant differences in the levels of IL-1RA, IL-10, GM-CSF, eotaxin, and IP-10 were observed between the HD-TAK and HD-GCA groups. Multiple logistic regression analysis demonstrated that only FGF-2 and IP-10 could significantly distinguish the diseases when added to age. Multiple logistic analysis using factors selected by the LASSO method revealed that FGF-2 was the only significant factor to distinguish the diseases when added to age. Among numerous cytokines and chemokines analyzed, only FGF-2 could be used together with age at diagnosis to differentiate TAK and GCA. Our results suggested the importance of considering the effects of age on serum cytokines..
10. Ayako Takaki-Kuwahara, Yojiro Arinobu, Kohta Miyawaki, Hisakata Yamada, Hirofumi Tsuzuki, Kensuke Irino, Masahiro Ayano, Yasutaka Kimoto, Hiroki Mitoma, Mitsuteru Akahoshi, Hiroshi Tsukamoto, Takahiko Horiuchi, Hiroaki Niiro, Koichi Akashi, CCR6+ group 3 innate lymphoid cells accumulate in inflamed joints in rheumatoid arthritis and produce Th17 cytokines, Arthritis Research and Therapy, 10.1186/s13075-019-1984-x, 21, 1, 2019.08.
11. Yoshifumi Tada, Satomi Inokuchi, Akihito Maruyama, Rie Suematsu, Mariko Sakai, Yuri Sadanaga, Nobuyuki Ono, Yojiro Arinobu, Syuichi Koarada, Are the 2016 EULAR/ACR/PRINTO classification criteria for macrophage activation syndrome applicable to patients with adult-onset Still’s disease?, Rheumatology International, 10.1007/s00296-018-4114-1, 39, 1, 97-104, 2019.01, The objectives of this study are to determine whether the 2016 European League Against Rheumatism/American College of Rheumatology/Paediatric Rheumatology International Trials Organization classification criteria for macrophage activation syndrome (MAS) complicating systemic juvenile idiopathic arthritis (SJIA) can be used to identify MAS in patients with adult-onset Still’s disease (AOSD). Using laboratory data from 76 AOSD patients with and without MAS, we analyzed the ability of the collective and individual constitutive elements of the 2016 MAS in SJIA criteria and additional laboratory measures to discriminate between AOSD patients with (n = 16) and without (n = 60) MAS. Cutoff values to determine the sensitivity, specificity, and predictive values were calculated from receiver operating characteristic curves, and modified classification criteria for MAS in AOSD were evaluated. The 2016 MAS in SJIA classification criteria had an overall sensitivity of 100%, specificity of 70.0%, positive predictive value of 47.1%, and negative predictive value of 100% to discriminate between AOSD patients with and without MAS based on laboratory data. Among the individual criteria, the sensitivity of triglycerides (46.7%) and the specificity of ferritin (15.0%) for MAS in AOSD were particularly low. The sensitivity and specificity for classifying MAS in AOSD patients were increased to 100 and 93%, respectively, by excluding triglycerides and changing the cutoff values for other criteria in the 2016 MAS in SJIA classification. The 2016 classification criteria for MAS in SJIA had higher sensitivity but lower specificity to identify MAS in AOSD patients compared with SJIA patients..
12. Masahiro Ayano, Hiroshi Tsukamoto, Hiroki Mitoma, Yasutaka Kimoto, Mitsuteru Akahoshi, Yojiro Arinobu, Toshihiro Miyamoto, Takahiko Horiuchi, Hiroaki Niiro, Koji Nagafuji, Mine Harada, Koichi Akashi, CD34-selected versus unmanipulated autologous haematopoietic stem cell transplantation in the treatment of severe systemic sclerosis
A post hoc analysis of a phase I/II clinical trial conducted in Japan, Arthritis Research and Therapy, 10.1186/s13075-019-1823-0, 21, 1, 2019.01, Background: The effectiveness of autologous haematopoietic stem cell transplantation (auto-HSCT) in treating severe systemic sclerosis (SSc) is established; however, the necessity of purified CD34+ cell grafts and the appropriate conditioning regimen remain unclear. This study aimed to compare the efficacy and safety of CD34-selected auto-HSCT with unmanipulated auto-HSCT to treat severe SSc. Methods: This study was a post hoc analysis of a phase I/II clinical trial conducted in Japan. Nineteen patients with severe SSc were enrolled. Peripheral blood stem cells (PBSCs) were mobilised with cyclophosphamide (4 g/m 2 ) and filgrastim (10 μg/kg/day). Following PBSC collection by apheresis, CD34+ cells were immunologically selected in 11 patients. All patients were treated with high-dose cyclophosphamide (200 mg/kg) monotherapy as a conditioning regimen and received CD34-selected (n = 11) or unmanipulated auto-HSCT (n = 8). Changes in skin sclerosis and pulmonary function were assessed over an 8-year follow-up period. Differences in the changes, toxicity, progression-free survival (PFS) and overall survival were compared between patients who had received CD34-selected auto-HSCT and those who had received unmanipulated auto-HSCT. Results: Skin sclerosis progressively improved after transplantation over an 8-year follow-up period in both groups, and the improvement was significantly greater in the CD34-selected group than in the unmanipulated group. Forced vital capacity in the CD34-selected group continuously increased over 8 years, whereas in the unmanipulated group it returned to baseline 3 years after transplantation. Toxicity and viral infections, such as cytomegalovirus infection and herpes zoster, were more frequently found in the CD34-selected group than in the unmanipulated group. The frequency of severe adverse events, such as bacterial infections or organ toxicity, was similar between the two groups. No treatment-related deaths occurred in either treatment group. PFS of the CD34-selected group was greater than that of the unmanipulated group, and the 5-year PFS rates of the CD34-selected and unmanipulated group were 81.8% and 50% respectively. Conclusions: CD34-selected auto-HSCT may produce favourable effects on improvement of skin sclerosis and pulmonary function compared with unmanipulated auto-HSCT. Use of CD34-selected auto-HSCT with high-dose cyclophosphamide monotherapy as a conditioning regimen may offer an excellent benefit-to-risk balance..
13. Ayako Nishino, Shin ya Kawashiri, Tomohiro Koga, Naoki Iwamoto, Kunihiro Ichinose, Mami Tamai, Hideki Nakamura, Tomoki Origuchi, Yukitaka Ueki, Tamami Yoshitama, Nobutaka Eiraku, Naoki Matsuoka, Akitomo Okada, Keita Fujikawa, Hiroaki Hamada, Tomomi Tsuru, Shuji Nagano, Yojiro Arinobu, Toshihiko Hidaka, Atsushi Kawakami, Ultrasonographic Efficacy of Biologic and Targeted Synthetic Disease-Modifying Antirheumatic Drug Therapy in Rheumatoid Arthritis From a Multicenter Rheumatoid Arthritis Ultrasound Prospective Cohort in Japan, Arthritis Care and Research, 10.1002/acr.23551, 70, 12, 1719-1726, 2018.12, Objective: To explore the variables associated with initial favorable power Doppler (PD) ultrasound (US) response induced by biologic and targeted synthetic disease-modifying antirheumatic drugs (b/tsDMARDs) in patients with rheumatoid arthritis (RA). Methods: We have been prospectively investigating the course of active RA patients using US after the introduction of b/tsDMARDs in the Kyushu region of Japan since June 2013. A total of 150 patients have completed the first 6 months of observation at present and have been evaluated. US was assessed in 22 joints of bilateral hands using gray-scale and PD images on a scale from 0–3. The sum of these scores was used as the indicator of US disease activity. We defined PD remission as attaining a total PD score of 0 at 6 months and investigated the associated variables by multivariate logistic regression analysis. Results: The total PD and gray-scale scores and the clinical composite measures significantly improved at 6 months, whereas these reductions were less in bDMARD switchers as compared with bDMARD-naive patients. Multivariate logistic regression analysis showed that short disease duration, the absence of any previous use of bDMARDs, and low total PD scores at baseline were independent predictors of PD remission at 6 months. Conclusion: This present prospective US cohort has for the first time shown the variables that are associated with initial PD response to b/tsDMARDs..
14. Yukimi Otsuka, Chikako Kiyohara, Yusuke Kashiwado, Takuya Sawabe, Shuji Nagano, Yasutaka Kimoto, Masahiro Ayano, Hiroki Mitoma, Mitsuteru Akahoshi, Yojiro Arinobu, Hiroaki Niiro, Koichi Akashi, Takahiko Horiuchi, Effects of tumor necrosis factor inhibitors and tocilizumab on the glycosylated hemoglobin levels in patients with rheumatoid arthritis; an observational study, PloS one, 10.1371/journal.pone.0196368, 13, 4, 2018.04, Rheumatoid arthritis (RA) and diabetes mellitus (DM) are associated with inflammation. We tried to investigate the influence of tumor necrosis factor inhibitors (TNFi) and tocilizumab (TCZ) on the glucose metabolism of RA patients. RA patients in whom treatment with TNFi or TCZ was initiated from 2008 to 2015 were studied based on their medical records. We analyzed patients whose glycosylated hemoglobin (HbA1c) levels were measured both before and 3 months after the initiation of these biologic agents. The association between HbA1c reduction and the treatment was evaluated. From 971 cases treated with these biologic agents, 221 cases whose medical records of HbA1c were available, were included (TNFi, n = 154; TCZ, n = 67). Both the TNFi and TCZ groups had significantly lower HbA1c values at 1 month and 3 months after the initiation of treatment (TNFi, p<0.001; TCZ, p<0.001). Although the pretreatment HbA1c values did not differ (TNFi, 6.2%; TCZ, 6.2%; p = 0.532), the 3-month treatment HbA1c values were lower (TNFi, 6.1%; TCZ, 5.8%; p = 0.010) and the changes in HbA1c (ΔHbA1c) were greater (TNFi, 0.1%; TCZ, 0.4%; <;0.001) in the TCZ group. The reduction of HbA1c—defined by the achievement of a ΔHbA1c of 0.5%—was associated with baseline diagnosis of diabetes mellitus, baseline diabetes treatment, hospitalization, medical change during the observation period, and TCZ. In the multivariate logistic regression analysis, TCZ was associated with the reduction of HbA1c in comparison to TNFi (adjusted OR = 5.59, 95% CI = 2.56–12.2; p<0.001). The HbA1c levels in RA patients were significantly lower after the initiation of TNFi or TCZ. Our study suggests that TCZ decreases the HbA1c levels in RA patients to a greater extent than TNFi..
15. Sho Ueda, Mitsuteru Akahoshi, Atsunobu Takeda, Yasushi Inoue, Aya Omoto, Masahiro Ayano, Yasutaka Kimoto, Hiroki Mitoma, Yojiro Arinobu, Hiroaki Niiro, Hiroshi Tsukamoto, Takahiko Horiuchi, Shin-Ichi Hikita, Takako Fukuhara, Tatsuro Ishibashi, Koh-Hei Sonoda, Koichi Akashi, Long-term efficacy of infliximab treatment and the predictors of treatment outcomes in patients with refractory uveitis associated with Behçet's disease, European journal of rheumatology, 10.5152/eurjrheum.2017.17068, 5, 1, 9-15, 2018.03, OBJECTIVE: To assess the long-term efficacy and safety of infliximab (IFX) treatment for refractory uveitis associated with Behçet's disease (BD) and to identify predictors of long-term IFX therapy outcomes.
METHODS: We retrospectively studied 44 consecutive BD patients with uveitis who were started on IFX therapy and analyzed the efficacy and safety of IFX and the treatment continuation rate. To determine predictors of IFX responsiveness, we analyzed the clinical characteristics of the patients who received regular maintenance therapy and those who required treatment intensification. The serum cytokine levels prior to IFX were measured through the Bio-Plex human cytokine assays.
RESULTS: IFX significantly reduced the frequency of ocular attacks and improved the visual acuity of patients with BD-related uveitis. However, approximately half of the patients required dose escalations, necessitating a shortening of the intervals between IFX infusions due to loss of efficacy during the 5-year treatment. The frequency of ocular attacks was significantly higher in patients with complete BD than in patients with incomplete BD. A multiplex cytokine analysis revealed that patients with BD-related uveitis exhibited increased serum IL-2, IL-6, IL-8, and MCP-1 levels. Moreover, among BD patients, the serum IL-2 and IL-6 levels were particularly high in those who maintained remission and received regular IFX treatments.
CONCLUSION: We confirmed the long-term efficacy and tolerability of IFX in patients with BD-related uveitis. Our results indicate that complete BD may be less responsive to IFX and that the pretreatment serum cytokine profiles may be useful for predicting the long-term IFX therapy outcomes..
16. Shinichi Mizuno, Tadafumi Iino, Hidetoshi Ozawa, Yojiro Arinobu, Yong Chong, Koichi Akashi, Notch1 expression is regulated at the post-transcriptional level by the 3′ untranslated region in hematopoietic stem cell development, International journal of hematology, 10.1007/s12185-017-2358-2, 107, 3, 311-319, 2018.03, In hematopoiesis, the expression of critical genes is regulated in a stage-specific manner to maintain normal hematopoiesis. Notch1 is an essential gene involved in the commitment and development of the T-cell lineage. However, the regulation of Notch1 in hematopoiesis is controversial, particularly at the level of hematopoietic stem cell (HSC). Here, we found that the expression of Notch1 is controlled at the post-transcriptional level in HSCs. HSCs express a considerable level of Notch1 mRNA, but its protein level is very low, suggesting a post-transcriptional suppression for Notch1. Using a retroviral sensor vector expressing a fusion mRNA of GFP and 3′ untranslated region (3′UTR) of a target gene, we demonstrated that the Notch1-3′UTR had a post-translational suppressive effect only at the HSC but not in the downstream progenitor stages. The sequence motif AUnA was required for this post-transcriptional regulation by the Notch1-3′UTR. Interestingly, this Notch1-3′UTR-mediated suppressive effect was relieved when HSCs were placed in the thymus, but not in the bone marrow. Thus, the expression of Notch1 in HSCs is regulated by microenvironment at the post-transcriptional level, which may control T lymphoid lineage commitment from HSCs..
17. Yuma Sakamoto, Takuaki Yamamoto, Nobuhiko Sugano, Daisuke Takahashi, Toshiyuki Watanabe, Takashi Atsumi, Junichi Nakamura, Yukiharu Hasegawa, Koichi Akashi, Ichiei Narita, Takeshi Miyamoto, Tsutomu Takeuchi, Katsunori Ikari, Koichi Amano, Atsuhiro Fujie, Toshikazu Kubo, Yoshifumi Tada, Ayumi Kaneuji, Hiroaki Nakamura, Tomoya Miyamura, Tamon Kabata, Ken Yamaji, Takahiro Okawa, Akihiro Sudo, Kenji Ohzono, Yoshiya Tanaka, Yuji Yasunaga, Shuichi Matsuda, Yuuki Imai, Yasuharu Nakashima, Goro Motomura, Satoshi Ikemura, Ryosuke Yamaguchi, Kazuyuki Karasuyama, Kazuhiko Sonoda, Takashi Nishii, Takashi Sakai, Masaki Takao, Tohru Irie, Tsuyoshi Asano, Norimasa Iwasaki, Tatsuya Atsumi, Satoshi Tamaoki, Ryosuke Nakanishi, Satoe Tanabe, Shunji Kishida, Shigeo Hagiwara, Taisuke Seki, Hiroshi Tsukamoto, Hiroaki Niiro, Yojiro Arinobu, Mitsuteru Akahoshi, Hiroshi Mitoma, Masahiro Ayano, Takeshi Kuroda, Yoshiaki Toyama, Atsushi Funayama, Hironari Hanaoka, Kunihiro Yamaoka, Yasushi Kawaguchi, Hisashi Yamanaka, Tetsuji Hosozawa, Shigeki Momohara, Kentaro Chino, Mikihiro Fujioka, Keichiro Ueshima, Masashi Ishida, Masazumi Saito, Shigeki Hayashi, Akira Ikegami, Toru Ichiseki, Shigekazu Mizokawa, Yoichi Ohta, Yoshitomo Kajino, Fumio Sekiya, Fujio Higuchi, Masahiro Hasegawa, Noriki Miyamoto, Shinichi Miyazaki, Toshio Yamaguchi, Wataru Ando, Kazuyoshi Saito, Kazuhisa Nakano, Yutaka Kuroda, Takuma Yamasaki, Masato Akiyama, Michiaki Kubo, Yoichiro Kamatani, Yukihide Iwamoto, Shiro Ikegawa, Genome-wide Association Study of Idiopathic Osteonecrosis of the Femoral Head, Scientific reports, 10.1038/s41598-017-14778-y, 7, 1, 2017.12, Idiopathic osteonecrosis of the femoral head (IONFH) is an ischemic disorder that causes bone necrosis of the femoral head, resulting in hip joint dysfunction. IONFH is a polygenic disease and steroid and alcohol have already known to increase its risk; however, the mechanism of IONFH remains to be elucidated. We performed a genome-wide association study using ~60,000 subjects and found two novel loci on chromosome 20q12 and 12q24. Big data analyses identified LINC01370 as a candidate susceptibility gene in the 20q12 locus. Stratified analysis by IONFH risk factors suggested that the 12q24 locus was associated with IONFH through drinking capacity. Our findings would shed new light on pathophysiology of IONFH..
18. Kohta Miyawaki, Hiromi Iwasaki, Takashi Jiromaru, Hirotake Kusumoto, Ayano Yurino, Takeshi Sugio, Yasufumi Uehara, Jun Odawara, Shinya Daitoku, Yuya Kunisaki, Yasuo Mori, Yojiro Arinobu, Hirofumi Tsuzuki, Yoshikane Kikushige, Tadafumi Iino, Koji Kato, Katsuto Takenaka, Toshihiro Miyamoto, Takahiro Maeda, Koichi Akashi, Identification of unipotent megakaryocyte progenitors in human hematopoiesis, Blood, 10.1182/blood-2016-09-741611, 129, 25, 3332-3343, 2017.06, The developmental pathway for human megakaryocytes remains unclear, and the definition of pure unipotent megakaryocyte progenitor is still controversial. Using single-cell transcriptome analysis, we have identified a cluster of cells within immature hematopoietic stem- and progenitor-cell populations that specifically expresses genes related to the megakaryocyte lineage. We used CD41 as a positive marker to identify these cells within the CD34+CD38+IL-3RαdimCD45RA- common myeloid progenitor (CMP) population. These cells lacked erythroid and granulocyte-macrophage potential but exhibited robust differentiation into the megakaryocyte lineage at a high frequency, both in vivo and in vitro. The efficiency and expansion potential of these cells exceeded those of conventional bipotent megakaryocyte/erythrocyte progenitors. Accordingly, the CD41+ CMP was defined as a unipotent megakaryocyte progenitor (MegP) that is likely to represent the major pathway for human megakaryopoiesis, independent of canonical megakaryocyte-erythroid lineage bifurcation. In the bone marrow of patients with essential thrombocythemia, the MegP population was significantly expanded in the context of a high burden of Janus kinase 2 mutations. Thus, the prospectively isolatable and functionally homogeneous human MegP will be useful for the elucidation of the mechanisms underlying normal and malignant human hematopoiesis. (Blood. 2017;129(25): 3332-3343)..
19. Tsuzuki H, Arinobu Y, Miyawaki K, Takaki A, Ota S, Ota Y, Mitoma H, Akahoshi M, Mori Y, Iwasaki H, Niiro H, Tsukamoto H, Akashi K, Functional interleukin-33 receptors are expressed in early progenitor stages of allergy-related granulocytes, Immunology, 150, 1, 64-73, 2017.01.
20. Hirofumi Tsuzuki, Yojiro Arinobu, Kohta Miyawaki, Ayako Takaki, Shun Ichiro Ota, Yuri Ota, Hiroki Mitoma, Mitsuteru Akahoshi, Yasuo Mori, Hiromi Iwasaki, Hiroaki Niiro, Hiroshi Tsukamoto, Koichi Akashi, Functional interleukin-33 receptors are expressed in early progenitor stages of allergy-related granulocytes, Immunology, 10.1111/imm.12667, 150, 1, 64-73, 2017.01, Interleukin-33 (IL-33) induces T helper type 2 (Th2) cytokine production and eosinophilia independently of acquired immunity, leading to innate immunity-mediated allergic inflammation. Allergy-related innate myeloid cells such as eosinophils, basophils and mast cells express the IL-33 receptor (IL-33R), but it is still unknown how IL-33 regulates allergic inflammation involving these cells and their progenitors. Here, we revealed that the functional IL-33R was expressed on eosinophil progenitors (EoPs), basophil progenitors (BaPs) and mast cell progenitors (MCPs). In the presence of IL-33, these progenitors did not expand, but produced a high amount of Th2 and pro-inflammatory cytokines such as IL-9, IL-13, IL-1β and IL-6. The amount of cytokines produced by these progenitors was greater than that by mature cells. In vivo, IL-33 stimulated the expansion of EoPs, but it was dependent upon the elevated serum IL-5 that is presumably derived from type 2 innate lymphoid cells that express functional IL-33R. These data collectively suggest that EoPs, BaPs and MCPs are not only the sources of allergy-related granulocytes, but can also be sources of allergy-related cytokines in IL-33-induced inflammation. Because such progenitors can differentiate into mature granulocytes at the site of inflammation, they are potential therapeutic targets in IL-33-related allergic diseases..
21. Yuri Ota, Hiroaki Niiro, Shun ichiro Ota, Naoko Ueki, Hirofumi Tsuzuki, Tsuyoshi Nakayama, Koji Mishima, Kazuhiko Higashioka, Siamak Jabbarzadeh-Tabrizi, Hiroki Mitoma, Mitsuteru Akahoshi, Yojiro Arinobu, Akiko Kukita, Hisakata Yamada, Hiroshi Tsukamoto, Koichi Akashi, Generation mechanism of RANKL+ effector memory B cells
Relevance to the pathogenesis of rheumatoid arthritis, Arthritis Research and Therapy, 10.1186/s13075-016-0957-6, 18, 1, 2016.03, Background: The efficacy of B cell-depleting therapies for rheumatoid arthritis underscores antibody-independent functions of effector B cells such as cognate T-B interactions and production of pro-inflammatory cytokines. Receptor activator of nuclear factor ΚB ligand (RANKL) is a key cytokine involved in bone destruction and is highly expressed in synovial fluid B cells in patients with rheumatoid arthritis. In this study we sought to clarify the generation mechanism of RANKL+ effector B cells and their impacts on osteoclast differentiation. Methods: Peripheral blood and synovial fluid B cells from healthy controls and patients with rheumatoid arthritis were isolated using cell sorter. mRNA expression of RANKL, osteoprotegerin, tumor necrosis factor (TNF)-α, and Blimp-1 was analyzed by quantitative real-time polymerase chain reaction. Levels of RANKL, CD80, CD86, and CXCR3 were analyzed using flow cytometry. Functional analysis of osteoclastogenesis was carried out in the co-culture system using macrophage RAW264 reporter cells. Results: RANKL expression was accentuated in CD80+CD86+ B cells, a highly activated B-cell subset more abundantly observed in patients with rheumatoid arthritis. Upon activation via B-cell receptor and CD40, switched-memory B cells predominantly expressed RANKL, which was further augmented by interferon-γ (IFN-γ) but suppressed by interleukin-21. Strikingly, IFN-γ also enhanced TNF-α expression, while it strongly suppressed osteoprotegerin expression in B cells. IFN-γ increased the generation of CXCR3+RANKL+ effector B cells, mimicking the synovial B cell phenotype in patients with rheumatoid arthritis. Finally, RANKL+ effector B cells in concert with TNF-α facilitated osteoclast differentiation in vitro. Conclusions: Our current findings have shed light on the generation mechanism of pathogenic RANKL+ effector B cells that would be an ideal therapeutic target for rheumatoid arthritis in the future..
22. Atsushi Tanaka, Hiroshi Tsukamoto, Hiroki Mitoma, Chikako Kiyohara, Naoyasu Ueda, Masahiro Ayano, Shun ichiro Ohta, Yasutaka Kimoto, Mitsuteru Akahoshi, Yojiro Arinobu, Hiroaki Niiro, Yoshifumi Tada, Takahiko Horiuchi, Koichi Akashi, Serum progranulin levels are elevated in dermatomyositis patients with acute interstitial lung disease, predicting prognosis, Arthritis Research and Therapy, 10.1186/s13075-015-0547-z, 17, 1, 2015.12, Introduction: Progranulin (PGRN), a pleiotropic growth factor, has emerged as an immunoregulatory molecule. Because the roles of PGRN in dermatomyositis (DM) are still unknown, we investigated whether serum PGRN levels are associated with disease activity and prognosis in DM patients, particularly in those with DM complicated with interstitial lung disease (ILD). Methods: The serum levels of PGRN were measured by enzyme-linked immunosorbent assay in patients with DM (n =57; acute/subacute interstitial pneumonia (A/SIP): n =17, chronic interstitial pneumonia (CIP): n =24, without ILD: n =16), polymyositis (PM, n =21; including 6 with ILD) and normal healthy controls (NHCs, n =60). We assessed the correlation between the serum PGRN levels and the activity indexes of ILD or prognosis in DM patients with ILD. Results: Serum PGRN levels were significantly higher in DM patients than in PM patients (P =0.0025) and in NHCs (P <0.0001). In DM patients, the levels were significantly higher in patients with A/SIP than in those with CIP (P <0.0001) or without ILD (P =0.0003). The serum PGRN levels in DM patients with ILD significantly correlated with serum ferritin (r S =0.77, P <0.0001), lactate dehydrogenase (r S =0.54, P =0.0003) and C-reactive protein (r S =0.48, P =0.0015) levels. Moreover, in DM patients with ILD, the cumulative survival rate for 6 months was significantly lower in the group with serum PGRN levels ≥200 ng/ml (67%) than in the group with serum PGRN levels <200 ng/ml (100%) (P =0.0009). Conclusions: Serum PGRN is associated with disease activity and prognosis of DM with ILD. PGRN may play a role in the pathogenesis of DM and could be a useful biomarker..
23. Masahiro Ayano, Hiroshi Tsukamoto, Kentaro Kohno, Naoyasu Ueda, Atsushi Tanaka, Hiroki Mitoma, Mitsuteru Akahoshi, Yojiro Arinobu, Hiroaki Niiro, Takahiko Horiuchi, Koichi Akashi, Increased CD226 expression on CD8+ T cells is associated with upregulated cytokine production and endothelial cell injury in patients with systemic sclerosis, Journal of Immunology, 10.4049/jimmunol.1403046, 195, 3, 892-900, 2015.08, Systemic sclerosis (SSc) is an autoimmune disease characterized by vascular damage and fibrosis of the skin and internal organs. Because activated and oligoclonally expanded CD8+ T cells can be detected in peripheral blood and lungs of SSc patients, effector memory CD8+ T cells may play a critical role for organ involvement in SSc; however, the pathogenic functions of effector memory CD8+ T cells remain incompletely understood. In this study, we performed DNA microarray analysis of the sort-purified effector memory CD8+ T cells from SSc patients and healthy controls, and showed that the expression of genes related to immune response and cell adhesion, including CD226 (also known as DNAX accessory molecule-1 [DNAM-1]), was significantly altered. Moreover, detailed analysis of CD226 revealed that CD226highCD8+ T cells were increased in SSc patients (mean, 50.7%) compared with healthy controls (32.9%) and were appreciably associated with the severity of skin sclerosis and interstitial lung disease. Furthermore, CD226+CD8+T cells produced higher amount of various cytokines than CD226- ones, and CD226highCD8+ T cells from SSc patients showed upregulated IL-13 production and positive correlation with the cytotoxic capacity of CD8+ T cells against HUVECs. Finally, the neutralization of CD226 in CD8+ T cells impaired costimulation, cytokine productions, and cytolysis against HUVECs. These findings indicate that upregulated CD226 expression on CD8+ T cells reflects disease severity and is involved in SSc pathogenesis via the production of various cytokines, including profibrotic IL-13 and endothelial cell injury, and that CD226 may be a useful target in the treatment of SSc..
24. Miyawaki K, Arinobu Y, Iwasaki H, Kohno K, Tsuzuki H, Iino T, Shima T, Kikushige Y,Takenaka K, Miyamoto T, Akashi K, CD41 marks the initial myelo-erythroid lineage specification in adult mouse hematopoiesis: redefinition of murine common myeloid progenitor., Stem Cells, 33: 976-987, 2015.03.
25. Kohta Miyawaki, Yojiro Arinobu, Hiromi Iwasaki, Kentaro Kohno, Hirofumi Tsuzuki, Tadafumi Iino, Takahiro Shima, Yoshikane Kikushige, Katsuto Takenaka, Toshihiro Miyamoto, Koichi Akashi, CD41 marks the initial myelo-erythroid lineage specification in adult mouse hematopoiesis
Redefinition of murine common myeloid progenitor, STEM CELLS, 10.1002/stem.1906, 33, 3, 976-987, 2015.03, Previous studies have predicted that reciprocal activation of GATA-1 and PU.1 regulates myelo-erythroid versus myelo-lymphoid lineage commitment in early hematopoiesis. Such PU.1-activating myelo-lymphoid progenitors exist within the lymphoid-primed multipotent progenitor (LMPP) population at the primitive Lineage-Sca-1+c-Kit+ (LSK) stage. We here show that the counterpart of GATA-1-activating myelo-erythroid progenitor resides also at the LSK stage, expressing CD41 at a high level. Purified CD41hi LSK cells showed exceedingly strong and prolonged myelo-erythroid-restricted reconstitution, and primed myelo-erythroid gene expression with a more primitive molecular signature as compared to the original common myeloid progenitor (CMP). The CD41hi LSK cells more strongly contributed to emergent and malignant myelopoiesis than LMPPs, and produced the original CMP by downregulating Sca-1 and CD41, suggesting that they are the earliest CMPs. Thus, the hematopoietic developmental map should be revised by integrating the primary branchpoint comprised of the new, isolatable CD41hi CMP and the LMPP populations. Stem Cells 2015;33:976-987.
26. 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, 1, 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 Sjögren'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..
27. Tsuyoshi Nakayama, Mitsuteru Akahoshi, Kensuke Irino, Yasutaka Kimoto, Yojiro Arinobu, Hiroaki Niiro, Hiroshi Tsukamoto, Takahiko Horiuchi, Koichi Akashi, Transient antiphospholipid syndrome associated with primary cytomegalovirus infection
a case report and literature review, Case reports in rheumatology, 10.1155/2014/271548, 2014, 271548, 2014, Viral infection is known to induce transient autoimmunity in humans. Acute cytomegalovirus (CMV) infection is implicated in occasional thrombosis formation. We here, for the first time, report a 19-year-old female who had an acute CMV infection, leading to a deep venous thrombosis and a pulmonary embolism along with transient appearance of lupus anticoagulant. The pathological role of antiphospholipid antibodies in CMV-mediated thrombosis is discussed..
28. Naoyasu Ueda, Hiroshi Tsukamoto, Hiroki Mitoma, Masahiro Ayano, Atsushi Tanaka, Shun Ichiro Ohta, Yasushi Inoue, Yojiro Arinobu, Hiroaki Niiro, Koichi Akashi, Takahiko Horiuchi, The cytotoxic effects of certolizumab pegol and golimumab mediated by transmembrane tumor necrosis factorα, Inflammatory bowel diseases, 10.1097/MIB.0b013e318280b169, 19, 6, 1224-1231, 2013.05, Background: Anti-tumor necrosis factor α (anti-TNF-α) agents have been successfully applied for the treatment of rheumatoid arthritis, Crohn's disease, and other chronic inflammatory diseases. Not only the neutralization of soluble TNF-α but also the effect on transmembrane TNF-α is important mechanisms of action of anti-TNF-α agents. This study investigated the cytotoxic effects of new anti-TNF-α agents, certolizumab pegol and golimumab, which are mediated by transmembrane TNF-α. Methods: Transmembrane TNF-α-expressing Jurkat T cells that did not express TNF receptors were used. The binding ability of each anti-TNF-α agent to transmembrane TNF-α, antibody-dependent cell-mediated cytotoxicity, complement-dependent cytotoxicity, and the apoptotic effect were examined. Results: Certolizumab pegol and golimumab bound to transmembrane TNF-α. Golimumab induced antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity, which was comparable to infliximab and adalimumab. However, certolizumab pegol did not induce antibody-dependent cell-mediated cytotoxicity or complement-dependent cytotoxicity. Certolizumab pegol directly induced nonapoptotic cell death in transmembrane TNF-α-expressing cells. Golimumab induced a weaker apoptotic effect than infliximab and adalimumab. Conclusions: The cytotoxic effects of anti-TNF-α agents on TNF-α-expressing cells are considered to be associated with the clinical effect of these agents on granulomatous diseases. The direct cytotoxic effect of certolizumab pegol on TNF-α-producing cells may contribute to its clinical efficacy in Crohn's disease. Golimumab may be less effective for granulomatous diseases..
29. Atsushi Tanaka, Hiroshi Tsukamoto, Hiroki Mitoma, Chikako Kiyohara, Naoyasu Ueda, Masahiro Ayano, Shun ichiro Ohta, Yasushi Inoue, Yojirou Arinobu, Hiroaki Niiro, Takahiko Horiuchi, Koichi Akashi, Serum progranulin levels are elevated in patients with systemic lupus erythematosus, reflecting disease activity, Arthritis Research and Therapy, 10.1186/ar4087, 14, 6, 2012.11, Introduction: Progranulin (PGRN) is the precursor of granulin (GRN), a soluble cofactor for toll-like receptor 9 (TLR9) signaling evoked by oligonucleotide (CpG)-DNA. Because TLR9 signaling plays an important role in systemic lupus erythematosus (SLE), we investigated whether PGRN is involved in the pathogenesis of SLE.Methods: We measured concentrations of serum PGRN and interleukin-6 (IL-6) with enzyme-linked immunosorbent assay (ELISA) in patients with SLE (n = 68) and in healthy controls (n = 60). We assessed the correlation between the serum PGRN levels and established disease-activity indexes. The sera from the patients with high PGRN titers (>80 ng/ml) at the initial evaluation were reevaluated after the disease was ameliorated by treatment. We also measured the IL-6 concentration secreted by peripheral blood mononuclear cells (PBMCs) incubated with (a) oligonucleotide (CpG-B) in the presence or absence of recombinant human PGRN (rhPGRN); and (b) lupus sera in the presence or absence of a neutralizing anti-PGRN antibody.Results: Serum PGRN levels were significantly higher in SLE patients than healthy controls. Their levels were significantly associated with activity of clinical symptoms. They also significantly correlated with values of clinical parameters, including the SLE Disease Activity Index and anti-double-stranded DNA antibody titers, and inversely with CH50, C3, and C4 levels. Moreover, serum PGRN levels significantly decreased after successful treatment of SLE. The rhPGRN significantly upregulated the production of IL-6 by PBMCs stimulated with CpG-B. Patients' sera stimulated production of IL-6 from PBMCs, which was significantly impaired by neutralization of PGRN. The serum PGRN levels significantly correlated with the serum IL-6 levels.Conclusions: Serum PGRN could be a useful biomarker for disease activity of SLE. PGRN may be involved in the pathogenesis of SLE partly by enhancing the TLR9 signaling..
30. Hiroaki Niiro, Siamak Jabbarzadeh-Tabrizi, Yoshikane Kikushige, Takahiro Shima, Kumiko Noda, Shun Ichiro Ota, Hirofumi Tsuzuki, Yasushi Inoue, Yojiro Arinobu, Hiromi Iwasaki, Shinji Shimoda, Eishi Baba, Hiroshi Tsukamoto, Takahiko Horiuchi, Tadayoshi Taniyama, Koichi Akashi, CIN85 is required for Cbl-mediated regulation of antigen receptor signaling in human B cells, Blood, 10.1182/blood-2011-04-351965, 119, 10, 2263-2273, 2012.03, The aberrant regulation of B-cell receptor (BCR) signaling allows unwanted B cells to persist, thereby potentially leading to autoimmunity and B-cell malignancies. Casitas B-lineage lymphoma (Cbl) proteins suppress BCR signaling; however, the molecular mechanisms that control Cbl function in human B cells remain unclear. Here, we demonstrate that CIN85 (c-Cbl interacting protein of 85 kDa) is constitutively associated with c-Cbl, Cbl-b, and B-cell linker in B cells. Experiments using CIN85-overexpressing and CIN85-knockdown B-cell lines revealed that CIN85 increased c-Cbl phosphorylation and inhibited BCR-induced calcium flux and phosphorylation of Syk and PLCγ2, whereas it did not affect BCR internalization. The Syk phosphorylation in CIN85-overexpressing and CIN85-knockdown cells was inversely correlated with the ubiquitination and degradation of Syk. Moreover, CIN85 knockdown in primary B cells enhanced BCR-induced survival and growth, and increased the expression of BcLxL, A1, cyclin D2, and myc. Following the stimulation of BCR and Toll-like receptor 9, B-cell differentiation-associated molecules were up-regulated in CIN85-knockdown cells. Together, these results suggest that CIN85 is required for Cbl-mediated regulation of BCR signaling and for downstream events such as survival, growth, and differentiation of human B cells..
31. Tetsuro Yamamoto, Takahiko Horiuchi, Hisaaki Miyahara, Shigeru Yoshizawa, Junichi Maehara, Eisuke Shono, Kazuto Takamura, Haruhisa MacHida, Kaoru Tsujioka, Takehiko Kaneko, Naoki Uemura, Kenichi Suzawa, Norihiko Inagaki, Noriko Umegaki, Yoshihiro Kasamatsu, Akihito Hara, Yojiro Arinobu, Yasushi Inoue, Hiroaki Niiro, Yoichiro Kashiwagai, Shin Ichi Harashima, Tomoko Tahira, Hiroshi Tsukamoto, Koichi Akashi, Hereditary angioedema in Japan
Genetic analysis of 13 unrelated cases, American Journal of the Medical Sciences, 10.1097/MAJ.0b013e31822bdb65, 343, 3, 210-214, 2012.03, Introduction: The molecular bases and clinical features of hereditary angioedema (HAE) have not been systematically documented in Japan or in other Asian countries. Thus, the authors researched the genetic and clinical characteristics of Japanese patients with HAE. Methods: The authors analyzed the CIINH gene for mutations in 13 unrelated Japanese patients with HAE by means of the polymerase chain reaction and nucleotide sequencing. In addition, the authors searched the literature from January 1969 to October 2010 on Japanese patients with HAE. Results: Seven of the mutations found were novel, including 4 missense mutations (8728T>G, 8831C>A, 16661T>G and 16885C>A), 2 frameshift mutations (2281-2350del70, 14158delT) and 1 large deletion (at least 1 kb-length deletion including exon 4), whereas 6 mutations had previously been reported in European populations. Conclusions: The genetic and clinical characteristics in Japanese patients with HAE may be similar to those in Western patients although our sample size is small and the authors identified 7 novel mutations..
32. Yojiro Arinobu, Koichi Akashi, [Myeloid lineage commitment from the hematopoietic stem cell]., Japanese Journal of Allergology, 60, 7, 817-822, 2011.07.
33. Tsukamoto H, Nagafuji K, Horiuchi T, Mitoma H, Niiro H, Arinobu Y, Inoue Y, To K, Miyamoto T, Iwasaki H, Teshima T, Harada M, Akashi K. , Analysis of immune reconstitution after autologous CD34+ stem/progenitor cell transplantation for systemic sclerosis: predominant reconstitution of Th1 CD4+ T cells., Rheumatology., 50: 944-952, 2011.05.
34. Hiroshi Tsukamoto, Koji Nagafuji, Takahiko Horiuchi, Hiroki Mitoma, Hiroaki Niiro, Yojiro Arinobu, Yasushi Inoue, Kentaro To, Toshihiro Miyamoto, Hiromi Iwasaki, Takanori Teshima, Mine Harada, Koichi Akashi, Analysis of immune reconstitution after autologous CD34 + stem/progenitor cell transplantation for systemic sclerosis
Predominant reconstitution of Th1 CD4 + t cells, Rheumatology, 10.1093/rheumatology/keq414, 50, 5, 944-952, 2011.05, Objective: The aim of this study is to evaluate the mechanism of long-term effect of autologous haematopoietic stem cell transplantation (ASCT) in treatment of SSc. Methods: Eleven patients (three males and eight females) with SSc were enrolled. Blood mononuclear cells were harvested after mobilization treatment with CYC and G-CSF. CD34 + haematopoietic stem/progenitor cell fractions were purified and cryopreserved. Patients were transplanted with >2 × 10/kg autologous CD34 + cells after high-dose CYC (50 mg/kg for 4 days) conditioning. Immune reconstitution was evaluated serially by analysing lymphocyte subpopulations for 36 months. Results: Progressive improvement of skin sclerosis has been observed for 3 years in most of the patients. The serum level of anti-Scl-70, an auto-antibody specific to SSc, was progressively decreased after ASCT. Improvement of skin sclerosis was significantly associated with the change in the serum anti-Scl-70 level after ASCT at 36 months. Serum levels of KL-6 and surfactant protein D, indicators for interstitial pneumonia activity, were also significantly decreased. The number of CD8+ T cells immediately recovered within a month after ASCT, while the number of CD4+ T cells remained low for >36 months post-transplant. The majority of CD4 + cells were memory but not naïve T cells, and regulatory CD4 + T cells were not recovered. Th1/Th2 ratio was significantly increased after ASCT. Conclusions: ASCT with purified CD34 + cells was effective in controlling the disease activity of SSc. Th1/Th2 ratio was significantly increased for at least 3 years after ASCT..
35. Naoyasu Ueda, Yasushi Inoue, Daisuke Himeji, Yoshiya Shimao, Kensuke Oryoji, Hiroki Mitoma, Yojiro Arinobu, Hiroaki Niiro, Hiroshi Tsukamoto, Takahiko Horiuchi, Akira Ueda, Koichi Akashi, Wegener's granulomatosis detected initially by integrated 18F-fluorodeoxyglucose positron emission tomography/computed tomography, Modern Rheumatology, 10.1007/s10165-009-0255-0, 20, 2, 205-209, 2010.04, Early diagnosis is crucial for effective treatment of Wegener's granulomatosis, although this disease shows only atypical symptoms in the primary stage. This report describes a patient suspected of having a malignancy based on integrated 18F-fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT), which showed increased uptake in pulmonary nodules and nasopharyngeal mucosa. Integrated PET/CT is therefore considered to be useful to confirm the distribution and determine the optimal site for biopsy..
36. Katsuta H, Akashi T, Katsuta R, Nagaya M, Kim D, Arinobu Y, Hara M, Bonner-Weir S, Sharma AJ, Akashi K, Weir GC. , Single pancreatic beta cells co-express multiple islet hormone genes in mice. , Diabetologia. , 53: 128-138, 2010.01.
37. H. Katsuta, T. Akashi, R. Katsuta, M. Nagaya, D. Kim, Y. Arinobu, M. Hara, S. Bonner-Weir, A. J. Sharma, K. Akashi, G. C. Weir, Single pancreatic beta cells co-express multiple islet hormone genes in mice, Diabetologia, 10.1007/s00125-009-1570-x, 53, 1, 128-138, 2010.01, Aims/hypothesis: It is widely accepted that production of insulin, glucagon, somatostatin and pancreatic polypeptide in islet cells is specific to beta, alpha, delta and pancreatic polypeptide cells, respectively. We examined whether beta cells express other genes encoding islet hormones. Methods: Nested RT-PCR was performed on single beta cells of transgenic mice with green fluorescent protein (GFP) driven by mouse insulin I promoter (MIP-GFP). Results: Only 55% of adult beta cells expressed the insulin gene alone, while others expressed two or more islet hormone genes; 4% expressed all four hormone genes. In embryonic and neonatal cells, 60% to 80% of GFP+ cells co-expressed pancreatic polypeptide and insulin genes in contrast to 29% in adult. To clarify cell fate, we conducted lineage tracing using rat insulin II promoter-cre mice crossed with reporter mice Gt(ROSA)26Sor-loxP-flanked STOP-cassette-GFP. All GFP+ cells expressed insulin I and II genes, and showed similar heterogeneity of co-expression to that seen in MIP-GFP mice. Although we report expression of other hormone genes in a significant proportion of beta cells, our lineage tracing results demonstrate that after inducing InsII (also known as Ins2) expression, beta cell progenitors do not redifferentiate to non-beta cells. Conclusions/interpretation: This study shows co-expression of multiple hormone genes in beta cells of adult mice as well as in embryos and neonates. This finding could: (1) represent residual expression from beta cell precursors; (2) result from alternative developmental pathways for beta cells; or (3) denote the differentiation potential of these cells. It may be linked to functional heterogeneity. This heterogeneity in gene expression may provide a means to characterise the functional, cellular and developmental heterogeneity seen in beta cells..
38. Yojiro Arinobu, Hiromi Iwasaki, Koichi Akashi, Origin of basophils and mast cells, Allergology International, 10.2332/allergolint.08-RAI-0067, 58, 1, 21-28, 2009.10.
39. Shinji Shimoda, Hiroshi Miyakawa, Minoru Nakamura, Hiromi Ishibashi, Kentaro Kikuchi, Hiroto Kita, Hiroaki Niiro, Youjirou Arinobu, Nobuyuki Ono, Ian R. Mackay, M. Eric Gershwin, Koichi Akashi, CD4 T-cell autoreactivity to the mitochondrial autoantigen PDC-E2 in AMA-negative primary biliary cirrhosis, Journal of Autoimmunity, 10.1016/j.jaut.2008.05.003, 31, 2, 110-115, 2008.09, Approximately 5% of patients with primary biliary cirrhosis (PBC) lack characteristic anti-mitochondrial antibodies (AMA). Yet clinically AMA+ and AMA- patients are similar. Using both AMA+ and AMA- patients, we quantitated the frequency of autoreactive T cells that respond to the major CD4 T-cell epitope, PDC-E2 163-176, using limiting dilution assays and quantitation of IFN-γ, IL-10 and IL-4. Further, based on data that both PBC patients and healthy subjects have CD4+ T cells that recognize PDC-E2 163-176 but with differing costimulation requirements, assays were performed using two different antigen-presenting cell (APC) systems: either autologous peripheral blood mononuclear cells (PBMC) or HLA DR53 transfected mouse fibroblast cell lines (L-DR53). When costimulation-incompetent L-DR53 were used as APCs, the PDC-E2 CD4 T-cell frequency and capacity for IFN-γ production were equivalent in both AMA+ and AMA- patients but the frequencies of such cells were significantly lower in normals, with IL-10 production similar in all three groups. Thus, in PBC there is 'universal' autoreactive CD4+ T-cell immune responsiveness to the critical autoantigen, PDC-E2. These observations emphasize that the mitochondrial autoreactivity in PBC is a multi-lineage response and hence, AMA-negative PBC may be an anachronism that refers only to sera autoantibodies..
40. Cantor AB, Iwasaki H, Arinobu Y, Moran TB, Shigematsu H, Sullivan MR, Akashi K, Orkin SH., Antagonism of FOG-1 and GATA factors in fate choice for the mast cell lineage. , J Exp Med. , 17: 611-624, 2008.03.
41. Alan B. Cantor, Hiromi Iwasaki, Yojiro Arinobu, Tyler B. Moran, Hirokazu Shigematsu, Matthew R. Sullivan, Koichi Akashi, Stuart H. Orkin, Antagonism of FOG-1 and GATA factors in fate choice for the mast cell lineage, Journal of Experimental Medicine, 10.1084/jem.20070544, 205, 3, 611-624, 2008.03, The zinc finger transcription factor GATA-1 requires direct physical interaction with the cofactor friend of GATA-1 (FOG-1) for its essential role in erythroid and megakaryocytic development. We show that in the mast cell lineage, GATA-1 functions completely independent of FOG proteins. Moreover, we demonstrate that FOG-1 antagonizes the fate choice of multipotential progenitor cells for the mast cell lineage, and that its downregulation is a prerequisite for mast cell development. Remarkably, ectopic expression of FOG-1 in committed mast cell progenitors redirects them into the erythroid, megakaryocytic, and granulocytic lineages. These lineage switches correlate with transcriptional down-regulation of GATA-2, an essential mast cell GATA factor, via switching of GATA-1 for GATA-2 at a key enhancer element upstream of the GATA-2 gene. These findings illustrate combinatorial control of cell fate identity by a transcription factor and its co factor, and highlight the role of transcriptional networks in lineage determination. They also provide evidence for lineage instability during early stages of hematopoietic lineage commitment. JEM.
42. Mitsuteru Akahoshi, Kumiko Aizawa, Shuji Nagano, Hisako Inoue, Atsushi Sadanaga, Yojiro Arinobu, Hiroaki Niiro, Hitoshi Nakashima, Acquired hemophilia in a patient with systemic lupus erythematosus
A case report and literature review, Modern Rheumatology, 10.1007/s10165-008-0084-6, 18, 5, 511-515, 2008, We report the case of a 38-year-old female patient with systemic lupus erythematosus (SLE) who developed acquired hemophilia caused by factor VIII (FVIII) inhibitors. She manifested spontaneous bleeding symptoms such as ecchymoses and hematuria. Laboratory findings showed an isolated prolongation of the activated partial thromboplastin time, reduced FVIII activity, and a high titer of FVIII inhibitors. She was successfully treated with oral predonisolone and cyclosporine in combination with steroid and cyclophosphamide pulse therapy..
43. Arinobu Y, Mizuno S, Chong Y, Shigematsu H, Iino T, Iwasaki H, Graf T, Mayfield R, Chan S, Kastner P, Akashi K. , Reciprocal activation of GATA-1 and PU.1 marks initial specification of hematopoietic stem cells into myeloerythroid and myelolymphoid lineages. , Cell Stem Cell , 1: 416-427, 2007.10.
44. Yojiro Arinobu, Shin ichi Mizuno, Yong Chong, Hirokazu Shigematsu, Tadafumi Iino, Hiromi Iwasaki, Thomas Graf, Robin Mayfield, Susan Chan, Philippe Kastner, Koichi Akashi, Reciprocal Activation of GATA-1 and PU.1 Marks Initial Specification of Hematopoietic Stem Cells into Myeloerythroid and Myelolymphoid Lineages, Cell stem cell, 10.1016/j.stem.2007.07.004, 1, 4, 416-427, 2007.10, A hierarchical hematopoietic development with myeloid versus lymphoid bifurcation has been proposed downstream of the multipotent progenitor (MPP) stage, based on prospective isolation of progenitors capable of generating only myeloerythroid cells (common myeloid progenitor, CMP) or only lymphocytes (common lymphoid progenitor, CLP). By utilizing GATA-1 and PU.1 transcription factor reporters, here we identified progenitor populations that are precursors for either CMPs or CLPs. Two independent populations expressing either GATA-1 or PU.1 resided within the CD34+Sca-1+c-Kit+ MPP fraction. The GATA-1+ MPP displayed potent myeloerythroid potential without giving rise to lymphocytes, whereas the PU.1+ MPP showed granulocyte/monocyte/lymphoid-restricted progenitor activity without megakaryocyte/erythroid differentiation. Furthermore, GATA-1+ and PU.1+ MPPs possessed huge expansion potential and differentiated into the original CMPs and CLPs, respectively. Thus, the reciprocal activation of GATA-1 and PU.1 primarily organizes the hematopoietic lineage fate decision to form the earliest hematopoietic branchpoint that comprises isolatable myeloerythroid and myelolymphoid progenitor populations..
45. Pilar Alcaide, Tatiana G. Jones, Graham M. Lord, Laurie H. Glimcher, Jenny Hallgren, Yojiro Arinobu, Koichi Akashi, Alison M. Paterson, Michael A. Gurish, Francis W. Luscinskas, Dendritic cell expression of the transcription factor T-bet regulates mast cell progenitor homing to mucosal tissue, Journal of Experimental Medicine, 10.1084/jem.20060626, 204, 2, 431-439, 2007.02, The transcription factor T-bet was identified in CD4+ T cells, and it controls interferon γ production and T helper type 1 cell differentiation. T-bet is expressed in certain other leukocytes, and we recently showed (Lord, G.M., R.M. Rao, H. Choe, B.M. Sullivan, A.H. Lichtman, F.W. Luscinskas, and L.H. Glimcher. 2005. Blood. 106:3432-3439) that it regulates T cell trafficking. We examined whether T-bet influences homing of mast cell progenitors (MCp) to peripheral tissues. Surprisingly, we found that MCp homing to the lung or small intestine in T-bet-/- mice is reduced. This is reproduced in adhesion studies using bone marrow-derived MCs (BMMCs) from T-bet-/- mice, which showed diminished adhesion to mucosal addresin cellular adhesion molecule-1 (MAdCAM-1) and vascular cell adhesion molecule-1 (VCAM-1), endothelial ligands required for MCp intestinal homing. MCp, their precursors, and BMMCs do not express T-bet, suggesting that T-bet plays an indirect role in homing. However, adoptive transfer experiments revealed that T-bet expression by BM cells is required for MCp homing to the intestine. Furthermore, transfer of WT BM-derived dendritic cells (DCs) to T-bet -/- mice restores normal MCp intestinal homing in vivo and MCp adhesion to MAdCAM-1 and VCAM-1 in vitro. Nonetheless, T-bet-/- mice respond vigorously to intestinal infection with Trichinella spiralis, eliminating a role for T-bet in MC recruitment to sites of infection and their activation and function. Therefore, remarkably, T-bet expression by DCs indirectly controls MCp homing to mucosal tissues. JEM.
46. Iwasaki H, Mizuno S, Arinobu Y, Ozawa H, Mori Y, Shigematsu H, Takatsu K, Tenen DG, Akashi K. , The order of expression of transcription factors directs hierarchical specification of hematopoietic lineages. , Genes Dev. , 20: 3010-3021, 2006.11.
47. Hiromi Iwasaki, Shin Ichi Mizuno, Yojiro Arinobu, Hidetoshi Ozawa, Yasuo Mori, Hirokazu Shigematsu, Kiyoshi Takatsu, Daniel G. Tenen, Koichi Akashi, The order of expression of transcription factors directs hierarchical specification of hematopoietic lineages, Genes and Development, 10.1101/gad.1493506, 20, 21, 3010-3021, 2006.11, The mechanism of lineage specification in multipotent stem cells has not been fully understood. We recently isolated progenitors with the eosinophil, basophil, or mast cell lineage potential, all of which originate from granulocyte/monocyte progenitors (GMPs). By using these prospectively purified progenitors, we show here that the expression timing of GATA-2 and CCAAT enhancer-binding protein α (C/EBPα) can differentially control their lineage commitment. The expression of GATA-2 instructed C/EBPα-expressing GMPs to commit exclusively into the eosinophil lineage, while it induced basophil and/or mast cell lineage commitment if C/EBPα was suppressed at the GMP stage. Furthermore, simply by switching the order of C/EBPα and GATA-2 transduction, even lymphoid-committed progenitors recaptured these developmental processes to be reprogrammed into each of these lineages. We propose that the order of expression of key transcription factors is critical for their interplay to selectively drive lineage specification programs, by which stem cells could generate multiple lineage cells in a hierarchical manner..
48. Arinobu Y, Iwasaki H, Gurish M, Mizuno S, Shigematsu H, Ozawa H, Tenen DG, Austen KF and Akashi K. , Developmental checkpoints of the basopohil/mast cell lineages in adult murine hematopoiesis. , Proc Natl Acad Sci USA. , 10.1073/pnas.0509148102, 102, 50, 18105-18110, 102: 18105-18110, 2005.12.
49. Hiromi Iwasaki, Chamorro Somoza, Hirokazu Shigematsu, Estelle A. Duprez, Junko Iwasaki-Arai, Shin Ichi Mizuno, Yojiro Arinobu, Kristin Geary, Pu Zhang, Tajhal Dayaram, Maris L. Fenyus, Shannon Elf, Susan Chan, Philippe Kastner, Claudia S. Huettner, Richard Murray, Daniel G. Tenen, Koichi Akashi, Distinctive and indispensable roles of PU.1 in maintenance of hematopoietic stem cells and their differentiation, Blood, 10.1182/blood-2005-03-0860, 106, 5, 1590-1600, 2005.09, The PU.1 transcription factor is a key regulator of hematopoietic development, but its role at each hematopoietic stage remains unclear. In particular, the expression of PU.1 in hematopoietic stem cells (HSCs) could simply represent "priming" of genes related to downstream myelolymphold lineages. By using a conditional PU.1 knock-out model, we here show that HSCs express PU.1, and its constitutive expression is necessary for maintenance of the HSC pool In the bone marrow. Bone marrow HSCs disrupted with PU. 1 m situ could not maintain hematopoiesis and were outcompeted by normal HSCs. PU.1-deficient HSCs also failed to generate the earliest myeloid and lymphold progenitors. PU.1 disruption in granulocyte/monocyte-committed progenitors blocked their maturation but not proliferation, resulting in myeloblast colony formation. PU. 1 disruption in common lymphoid progenitors, however, did not prevent their B-cell maturation. In vivo disruption of PU.1 in mature B cells by the CD19-Cre locus did not affect B-cell maturation, and PU.1-deficient mature B cells displayed normal proliferation in response to mitogenic signals including the cross-linking of surface immunoglobulln M (IgM). Thus, PU.1 plays indispensable and distinct roles in hematopoietic development through supporting HSC self-renewal as well as commitment and maturation of myeloid and lymphoid lineages..
50. Iwasaki H, Mizuno S, Mayfield R, Shigematsu H, Arinobu Y, Seed B, Gurish MF, Takatsu K, and Akashi K. , Identification of eosinophil lineage-committed progenitors in the murine bone marrow. , J Exp Med. , 10.1084/jem.20050548, 201, 12, 1891-1897, 201: 1891-1897, 2005.06.
51. Takeshi Inukai, Toshiya Inaba, Jinjun Dang, Ryoko Kuribara, Keiya Ozawa, Atsushi Miyajima, Wenshu Wu, A. Thomas Look, Yojiro Arinobu, Hiromi Iwasaki, Koichi Akashi, Keiko Kagami, Kumiko Goi, Kanji Sugita, Shinpei Nakazawa, TEF, an antiapoptotic bZIP transcription factor related to the oncogenic E2A-HLF chimera, inhibits cell growth by down-regulating expression of the common β chain of cytokine receptors, Blood, 10.1182/blood-2004-08-2976, 105, 11, 4437-4444, 2005.06, Gain and/or loss of function mediated by chimeric transcription factors generated by nonrandom translocations in leukemia is a key to understanding oncogenesis. E2A-hepatic leukemia factor (HLF), a chimeric basic region/leucine zipper (bZIP) transcription factor expressed in t(17;19)-positive leukemia cells, contributes to leukemogenesis through its potential to inhibit apoptosis. To identify physiologic counterparts of this chimera, we investigated the function of other bZIP factors that bind to the same DNA sequence recognized by E2A-HLF. Here, we show that thyrotroph embryonic factor (TEF), which shares a high level of sequence identity with HLF and recognizes the same DNA sequence, is expressed in a small fraction of each subset of hematolymphoid progenitors. When TEF was introduced into FL5.12 interleukin 3 (IL-3)-dependent cells, TEF protected the cells from apoptosis due to IL-3 deprivation. Unexpectedly, TEF also almost completely down-regulated expression of the common β (βc) chain of cytokine receptors. Consequently, TEF-expressing cells accumulated in G0/G1 phase without undergoing apoptosis. These findings suggest that TEF is one of the apoptotic regulators in hematopoietic progenitors and controls hematopoietic-cell proliferation by regulating the expression of the βc chain. In contrast, E2A-HLF promoted cell survival more efficiently than TEF but did not down-regulate βc chain expression, suggesting that E2A-HLF retains ideal properties for driving leukemic transformation..
52. Yasushi Inoue, Takeshi Otsuka, Hiroaki Niiro, Shuji Nagano, Yojirou Arinobu, Eiichi Ogami, Mitsuteru Akahoshi, Katsuhisa Miyake, Ichiro Ninomiya, Sakiko Shimizu, Hitoshi Nakashima, Mine Harada, Novel Regulatory Mechanisms of CD40-Induced Prostanoid Synthesis by IL-4 and IL-10 in Human Monocytes, Journal of Immunology, 10.4049/jimmunol.172.4.2147, 172, 4, 2147-2154, 2004.02, Interleukins IL-4 and IL-10 are considered to be central regulators for the limitation and eventual termination of inflammatory responses in vivo, based on their potent anti-inflammatory effects toward LPS-stimulated monocytes/macrophages and neutrophils. However, their role in T cell-dependent inflammatory responses has not been fully elucidated. In this study, we investigated the effects of both cytokines on the production of PGE 2, a key molecule of various inflammatory conditions, in CD40-stimulated human peripheral blood monocytes. CD40 ligation of monocytes induced the synthesis of a significant amount of PGE2 via inducible expression of the cyclooxygenase (COX)-2 gene. Both IL-10 and IL-4 significantly inhibited PGE2 production and COX-2 expression in CD40-stimulated monocytes. Using specific inhibitors for extracellular signal-related kinase (ERK) and p38 mitogen-activated protein kinase (MAPK), we found that both kinase pathways are involved in CD40-induced COX-2 expression. CD40 ligation also resulted in the activation of NF-κB Additional experiments exhibited that CD40 clearly induced the activation of the upstream kinases MAPK/ERK kinase 1/2, MAPK kinase 3/6, and I-κB in monocytes. IL-10 significantly inhibited CD40-induced activation of the ERK, p38 MAPK, and NF-κB pathways; however, inhibition by IL-4 was limited to the ERK pathway in monocytes. Neither IL-10 nor IL-4 affected the recruitment of TNFR-associated factors 2 and 3 to CD40 in monocytes. Collectively, IL-10 and IL-4 use novel regulatory mechanisms for CD40-induced prostanoid synthesis in monocytes, thus suggesting a potential role for these cytokines in regulating T cell-induced inflammatory responses, including autoimmune diseases..
53. Ayako Oguma, Takeshi Kojima, Daisuke Himeji, Yojiro Arinobu, Ikuo Kikuchi, Akira Ueda, A case of Legionella pneumonia complicated with acute respiratory distress syndrome treated with methylprednisolone and sivelestat sodium in combination with intravenous erythromycin and ciprofloxacin, Nihon Kokyūki Gakkai zasshi = the journal of the Japanese Respiratory Society, 42, 11, 956-960, 2004.01, A 64-year-old man was referred to us because of pneumonia refractory to panipenem/betamipron. His chest radiography showed patchy consolidations in the lower lobe of the right lung and in the middle field of the left lung, and severe hypoxia was present. He was diagnosed as having acute respiratory distress syndrome due to severe pneumonia, and was treated with pulse methylprednisolone and sivelestat sodium in combination with intravenous erythromycin and ciprofloxacin. The patient recovered with this treatment. Serological examination using blood samples collected on the 12th and 28th hospital days revealed elevation of anti-L. pneumophila serogroup I antibody. It is suggested that administration of methylprednisolone and sivelestat sodium in combination with intravenous erythromycin and ciprofloxacin in a case of severe Legionella pneumonia complicated with acute respiratory distress syndrome is effective, and may be of use in similar cases..
54. Shuji Nagano, Takeshi Otsuka, Hiroaki Niiro, Kunihiro Yamaoka, Yojirou Arinobu, Eiichi Ogami, Mitsuteru Akahoshi, Yasushi Inoue, Katsuhisa Miyake, Hitoshi Nakashima, Yoshiyuki Niho, Mine Harada, Molecular mechanisms of lipopolysaccharide-induced cyclooxygenase-2 expression in human neutrophils
Involvement of the mitogen-activated protein kinase pathway and regulation by anti-inflammatory cytokines, International immunology, 10.1093/intimm/dxf038, 14, 7, 733-740, 2002, Neutrophils are an important cellular source of proinflammatory mediators, whose regulation may be of potential benefit for the treatment of a number of inflammatory diseases. However, the mechanisms of lipopolysaccharide (LPS)-induced neutrophil activation and its regulation by anti-inflammatory cytokines have not yet been fully elucidated. Recent studies have revealed that mitogen-activated protein kinases (MAPK) play a crucial role in the generation of proinflammatory mediators in some cell types. Therefore, we conducted this study to determine whether MAPK activation could be involved in prostaglandin E2 (PGE2) production and cyclooxygenase (COX)-2 expression in LPS-stimulated human neutrophils. PD98059 (MEK1 inhibitor) and SB203580 (p38MAPK inhibitor) reduced PGE2 production as well as COX-2 expression in LPS-stimulated neutrophils. In addition, both extracellular signal-regulated protein kinase (ERK) and p38MAPK were phosphorylated and activated in time- and dose-dependent manners. Since we previously showed that IL-10 and IL-4 similarly inhibited COX-2 expression in LPS-stimulated neutrophils, we next tested the effects of IL-10 and IL-4 on the phosphorylation and activation of both kinases. IL-10 inhibited the phosphorylation and activation of p38MAPK, but not ERK. In addition, IL-4 caused a marginal inhibition in the activation of p38MAPK. Taken together, these results suggest that both ERK and p38MAPK pathways are involved in LPS-induced COX-2 expression and PGE2 production in neutrophils, and IL-10 and IL-4 inhibit neutrophil prostanoid synthesis by down-regulating the activation of p38MAPK..
55. Chika Morita, Takahiko Horiuchi, Hiroshi Tsukamoto, Nobuaki Hatta, Yuji Kikuchi, Yojiro Arinobu, Takeshi Otsuka, Takuya Sawabe, Shin Ichi Harashima, Kohei Nagasawa, Yoshiyuki Niho, Association of tumor necrosis factor receptor type II polymorphism 196R with systemic lupus erythematosus in the Japanese molecular and functional analysis, Arthritis and rheumatism, 10.1002/1529-0131(200112)44:12<2819::AID-ART469>3.0.CO;2-2, 44, 12, 2819-2827, 2001, Objective. To investigate whether a polymorphism(s) or mutation(s) in the tumor necrosis factor receptor II (TNFRII) gene is involved in the pathogenesis of systemic lupus erythematosus (SLE). Methods. All 10 exons of the TNFRII gene were analyzed by exon-specific polymerase chain reaction-single-strand conformation polymorphism, followed by nucleotide sequencing of exons that displayed aberrant bands. To analyze the function of the TNFRII polymorphisms, the full-length TNFRII complementary DNA of each allele was transfected in HeLa cells and then studied for specific binding of 125I-TNFα, as well as interleukin-6 (IL-6) production and cytotoxic activity after treatment with recombinant human TNFα. Results. We identified 4 polymorphisms, at codons 56, 181, 196, and 232. The latter 2 had amino acid substitutions M196R and E232K, respectively. Only the 196R allele was significantly associated with SLE in our 105 Japanese SLE patients, with an allele frequency of 20.5%, compared with 12.6% in 99 healthy controls (P = 0.0335). More importantly, using TNFRII-transfected HeLa cells, we demonstrated significantly increased IL-6 production by 196R TNFRII compared with 196M TNFRII. The cytotoxic activity induced by 196R TNFRII was also increased compared with that of 196M TNFRII. This increase was achieved without affecting the binding affinity of TNFα to TNFRII, as demonstrated by the finding that specific TNFα binding to the HeLa transfectants of 196R and 196M TNFRII was similar, with Kd values of 3.12 × 10-10M and 4.34 × 10-10M, respectively. Conclusion. These results suggest that 196R TNFRII, which transduces the signals of TNFα more effectively than does 196M TNFRII, is involved in the pathogenesis of SLE..
56. Yojiro Arinobu, Rie Sugimoto, Mina Akaiwa, Kazuhiko Arima, Takeshi Otsuka, Naotaka Hamasaki, Kenji Izuhara, Augmentation of signal transducer and activation of transcription (STAT)6 and STAT3 expression in stimulated B and T cells, Biochemical and Biophysical Research Communications, 10.1006/bbrc.2000.3674, 277, 2, 317-324, 2000.10.
57. P. S. Gao, X. Q. Mao, M. H. Roberts, Y. Arinobu, M. Akaiwa, T. Enomoto, Y. Dake, M. Kawai, S. Sasaki, N. Hamasaki, K. Izuhara, T. Shirakawa, J. M. Hopkin, Variants of STAT6 (signal transducer and activator of transcription 6) in atopic asthma, Journal of medical genetics, 37, 5, 380-382, 2000.05.
58. A. Heinzmann, X. Q. Mao, M. Akaiwa, R. T. Kreomer, P. S. Gao, K. Ohshima, R. Umeshita, Y. Abe, S. Braun, T. Yamashita, M. H. Roberts, R. Sugimoto, K. Arima, Y. Arinobu, B. Yu, S. Kruse, T. Enomoto, Y. Dake, M. Kawai, S. Shimazu, S. Sasaki, C. N. Adra, M. Kitaichi, H. Inoue, K. Yamauchi, N. Tomichi, F. Kurimoto, N. Hamasaki, J. M. Hopkin, K. Izuhara, T. Shirakawa, K. A. Deichmann, Genetic variants of IL-13 signalling and human asthma and atopy, Human molecular genetics, 10.1093/hmg/9.4.549, 9, 4, 549-559, 2000.03, Asthma and atopy show epidemiological association and are biologically linked by T-helper type 2 (T(h)2) cytokine-driven inflammatory mechanisms. IL-4 operates through the IL-4 receptor (IL-4R, a heterodimer of IL-4Rα and either γc or IL-13Rα1) and IL-13 operates through IL-13R (a heterodimer of IL-4Ra and IL-13Rα1) to promote IgE synthesis and IgE-based mucosal inflammation which typify atopy. Recent animal model data suggest that IL-13 is a central cytokine in promoting asthma, through the stimulation of bronchial epithelial mucus secretion and smooth muscle hyper-reactivity. We investigated the role of common genetic variants of IL-13 and IL-13Rα1 in human asthma, considering IgE levels. A novel variant of human IL-13, Gln110Arg, on chromosome 5q31, associated with asthma rather than IgE levels in case-control populations from Britain and Japan [peak odds ratio (OR) = 2.31, 95% CI 1.33-4.00]; the variant also predicted asthma and higher serum IL-13 levels in a general, Japanese paediatric population. Immunohistochemistry demonstrated that both subunits of IL-13R are prominently expressed in bronchial epithelium and smooth muscle from asthmatic subjects. Detailed molecular modelling analyses indicate that residue 110 of IL-13, the site of the charge-modifying variants Arg and Gln, is important in the internal constitution of the ligand and crucial in ligand-receptor interaction. A non-coding variant of IL-13Rα1, A1398G, on chromosome Xq13, associated primarily with high IgE levels (OR = 3.38 in males, 1.10 in females) rather than asthma. Thus, certain variants of IL-13 signalling are likely to be important promoters of human asthma; detailed functional analysis of their actions is needed..
59. H. Nakashima, M. Akahoshi, Y. Tanaka, K. Yamaoka, E. Ogami, S. Nagano, Y. Arinobu, H. Niiro, T. Otsuka, Y. Niho, Polymorphisms within the interleukin-10 receptor cDNA gene (IL10R) in Japanese patients with systemic lupus erythematosus, Rheumatology, 10.1093/rheumatology/38.11.1142, 38, 11, 1142-1144, 1999.11, Objective. To assess the association between polymorphisms within the interleukin-10 receptor cDNA gene (IL10R) and systemic erythematosus (SLE) in Japanese people. Method. We examined the IL-10 receptor genotype of 109 SLE patients and 102 healthy subjects by the reverse transcription-polymerase chain reaction-restriction fragment length polymorphism (RT-PCR-RFLP) method. Results. There was no difference in the IL10R genotype frequencies of these two groups. Conclusion. The IL10R genotype does not determine susceptibility to SLE in Japanese people..
60. H. Nakashima, T. Otsuka, Y. Ohba, M. Akahoshi, S. Nagano, E. Ogami, Y. Arinobu, K. Miyake, Y. Inoue, H. Niiro, Y. Kaji, Y. Niho, Two polymorphisms within interleukin-3 (hlL3) gene detected by mismatch PCR/RFLP, Genes and Immunity, 10.1038/sj.gene.6363636, 1, 2, 156-158, 1999.11, Two alleles of IL-3 have been reported to GenBank (GenBank M14743, M20137). The sequence difference between these two alleles is at the first nucleotide of the 27th codon (the 131st nucleotide from the initiation site): thymine and cytosine, and leading the amino acid difference: proline and serine (Pro27Ser). The other allelism, thymine and cytosine, was also observed at position -16 of the IL-3 upstream promotor region (GenBank L10616, M60870). We clarified that these substitutions were frequent polymorphisms in the Japanese population by using the mismatch-PCR (polymerase chain reaction)/RFLP (restriction fragment length polymorphism) method..
61. Mitsuteru Akahoshi, Hitoshi Nakashima, Yosuke Tanaka, Tsutomu Kohsaka, Shuji Nagano, Eiichi Ohgami, Yojiro Arinobu, Kunihiro Yamaoka, Hiroaki Niiro, Michiya Shinozaki, Hideki Hirakata, Takahiko Horiuchi, Takeshi Otsuka, Yoshiyuki Niho, Th1/Th2 balance of peripheral T helper cells in systemic lupus erythematosus, Arthritis and rheumatism, 10.1002/1529-0131(199908)42:8<1644::AID-ANR12>3.0.CO;2-L, 42, 8, 1644-1648, 1999.08, Objective. To analyze the Th1/Th2 balance of peripheral Th cells in patients with systemic lupus erythematosus (SLE). Methods. The Th1:Th2 ratio was analyzed in 3 groups: SLE without proteinuria (group I; n = 23), SLE with proteinuria (group II; n = 31), and normal controls (group III; n = 24). Group II patients who had undergone renal biopsy were classified into 3 subgroups based on their renal histopathologic findings. The intracellular cytokine detection method with flow cytometry was used to quantitate Th1 and Th2 cells. Results. There was no difference in the mean Th1:Th2 ratio between SLE patients (groups I and II) and healthy controls (group III). However, the mean value in group II was significantly higher than those in groups I and III. Moreover, within group II, the mean value in SLE patients who had diffuse proliferative lupus nephritis (World Health Organization class IV) was especially high. Conclusion. Although SLE has been considered to be a disease in which Th2 cells predominate, the Th1/Th2 balance of peripheral Th cells in SLE patients in the present study did not show a predominance of these cells. In contrast, among SLE patients with WHO class IV lupus nephritis, there was a strong predominance of Th1..
62. Hitoshi Nakashima, Hisako Inoue, Mitsuteru Akahoshi, Yosuke Tanaka, Kunihiro Yamaoka, Eiichi Ogami, Shuuji Nagano, Yojiro Arinobu, Hiroaki Niiro, Takeshi Otsuka, Yoshiyuki Niho, The combination of polymorphisms within interferon-γ receptor I and receptor 2 associated with the risk of systemic lupus erythematosus, FEBS Letters, 10.1016/S0014-5793(99)00701-2, 453, 1-2, 187-190, 1999.06, Genetic factors seem to play a significant role in susceptibility to systemic lupus erythematosus (SLE). We previously described the amino acid polymorphism (Val14Met) within the IFN-γ receptor 1 (IFN-γR1), and that the frequency of the Met14 allele in SLE patients was significantly higher than that of the healthy control population. We also found an amino acid polymorphism (Gln64Arg) within IFN-γ receptor 2 (IFN-γR2). Since the IFN-γ receptor is a complex consisting of IFN-γR1 and IFN-γR2, we searched for the particular combination of two kinds of amino acid polymorphisms found within the IFN-γ receptor which plays a prominent role in susceptibility to SLE. The greatest risk of the development of SLE was detected in the individuals who had the combination of IFNGR1 Met14/Val14 genotype and IFNGR2 Gln64/Gln64 genotype..
63. Arinobu Y, Atamas SP, Otsuka T, Niiro H, Yamaoka K, Mitsuyasu H, Niho Y, Hamasaki N, White B, Izuhara K., Antagonistic effects of an alternative splice variant of human IL-4, IL-4δ2 , on IL-4 activities in human monocytes and B cells. , Cell Immunol. , 10.1006/cimm.1998.1431, 191, 2, 161-167, 191: 161-167, 1999.02.
64. Hiromichi Mitsuyasu, Yukiyoshi Yanagihara, Xiao Quan Mao, Pei Sun Gao, Yojiro Arinobu, Kenji Ihara, Akira Takabayashi, Toshiro Hara, Tadao Enomoto, Sei Sasaki, Minoru Kawai, Naotaka Hamasaki, Taro Shirakawa, Julian M. Hopkin, Kenji Izuhara, Cutting edge
Dominant effect of Ile50Val variant of the human IL-4 receptor α-chain in IgE synthesis, Journal of Immunology, 162, 3, 1227-1231, 1999.02, Two variant of the IL-4R α-chain (IL-4Rα) gene have been recently identified in association with different atopic disorders. To clarify the etiological relationship between the two variants, we analyzed responsiveness to IL-4 of transfectants with four kinds of IL-4Rα carrying either Val or Ile at 50 and either Gln or Arg at 551. The substitution of Ile for Val augmented STAT6 activation, proliferation, and transcription activity of the Iε promoter by IL-4, whereas that of Arg for Gln did not change these IL-4 signals. Arg551 was not associated with atopic asthma in the Japanese population. CD23 expression and IgE synthesis by IL-4 were augmented in Ile50-bearing PBMC, compared with those bearing Val50. Taken together, substitution of Arg551 does not enhance the IL-4 signal for generation of germline ε transcript, whereas the substitution of Ile50 contributes to enhancement of IgE synthesis..
65. K. Izuhara, Y. Arinobu, H. Sumimoto, H. Nunoi, R. Takeya, K. Higuchi, K. Takeshige, N. Hamasaki, N. Harada, Association of the interleukin-4 receptor α chain with p47(phox), an activator of the phagocyte NADPH oxidase in B cells, Molecular Immunology, 10.1016/S0161-5890(98)00111-4, 36, 1, 45-52, 1999.01, Interleukin (IL)-4 plays an important role in IgE synthesis in B cells and in Th2 differentiation in T cells, IL-4 conducts its biological activities through binding to the IL-4 receptor (IL-4R) on the surface of target cells. IL-4R are thought to be composed of the IL-4R α chain (IL- 4Rα) and either the IL-2R γ chain or the IL-13R α chain. We have previously shown that the membrane-proximal portion in the cytoplasmic domain of the human IL-4Rα (hIL-4Rα) is critical for proliferation, generation of germline ε transcript, and activation of STAT6, based on analyses of truncated hIL-4Rαs. In this study, we found that p47p(phox), an activator of the phagocyte NADPH oxidase, binds to this portion by the two-hybrid system. Furthermore, we observed the association of p47(phox) with the hIL-4Rα in B cells derived from a normal donor. These results suggest that p47(phox) is involved in the signal transduction of IL-4 in B cells. However, activation of STAT6, CD23 expression, and IgE synthesis induced by IL-4 were not affected in p47(phox)-deficient patients, which raises the possibility that p47(phox) may be important in other signaling activities as well in B cells..
66. Hiroaki Niiro, Takeshi Otsuka, Eiichi Ogami, Kunihiro Yamaoka, Shuji Nagano, Mitsuteru Akahoshi, Hitoshi Nakashima, Yojiro Arinobu, Kenji Izuhara, Yoshiyuki Niho, MAP kinase pathways as a route for regulatory mechanisms of IL-10 and IL-4 which inhibit COX-2 expression in human monocytes, Biochemical and Biophysical Research Communications, 10.1006/bbrc.1998.9287, 250, 2, 200-205, 1998.09, Mitogen-activated protein kinases (MAPKs) are activated by various extracellular stimuli and play an important role in regulating the expression of proinflammatory molecules in monocytes/macrophages. We first questioned whether MAPK activation is involved in cyclooxygenase (COX)-2 expression in lipopolysaccharide (LPS)-stimulated human monocytes. LPS induced the expression of COX-2 protein and COX-2 mRNA as well as the phosphorylation and activation of extracellular signal-regulated protein kinase (ERK)2 and p38 MAPK in monocytes. The induction of COX-2 mRNA, COX-2 protein, and prostaglandin (PG)E2 by LPS was inhibited by the specific inhibitors of ERK and p38 MAPK, suggesting that the activation of ERK2 and p38 MAPK is involved in COX-2 expression in LPS-stimulated monocytes. Since we previously showed that interleukin (IL)-10 and IL-4 similarly inhibited COX-2 expression in LPS-stimulated monocytes, we next questioned whether these cytokines regulate the phosphorylation and activation of ERK2 and p38 MAPK in LPS-stimulated monocytes. Interestingly, LPS-induced phosphorylation and activation of ERK2 was significantly inhibited by IL-4 and IL-10, while that of p38 MAPK was inhibited by IL-10, but not IL-4. These results suggest that the mechanisms of inhibition by IL-10 and IL-4 of the LPS-induced expression of proinflammatory molecules could be ascribed to the regulatory effects of both cytokines on MAPK activation..
67. Mitsuyasu H, Izuhara K, Mao XQ, Gao PS, Arinobu Y, Enomoto T, Kawai M, Sasaki S, Dake Y, Hamasaki N, Shirakawa T and Hopkin JM. , Ile50Val variant of IL4R alpha upregulates IgE synthesis and associates with atopic ashma. , Nat Genet. , 10.1038/472, 19, 2, 119-120, 19: 119-120, 1998.06.
68. Kunihiro Yamaoka, Takeshi Otsuka, Hiroaki Niiro, Yojiro Arinobu, Yoshiyuki Niho, Naotaka Hamasaki, Kenji Izuhara, Activation of STAT5 by lipopolysaccharide through granulocyte-macrophage colony-stimulating factor production in human monocytes, Journal of Immunology, 160, 2, 838-845, 1998.01, LPS is a potent stimulator of monocytes, inducing many of their functions. Although the details of how LPS exerts such functions remain largely unknown, transcription factors such as nuclear factor-κB, nuclear factor-IL-6, and activator protein-1 have been shown to be involved in this process. However, to date it has been thought that no known STAT molecule plays a role in the activation of monocytes by LPS. In this study we examined whether some known STAT molecule is stimulated by LPS, based on the finding that a GAS motif sequence is conserved in the promoter regions of human, mouse, and rat cyclo-oxygenase-2 (COX-2) genes. Consequently, LPS induced activation of STAT5 in human monocytes, and this STAT5 activation occurred in an indirect way via granulocyte-macrophage CSF (GM-CSF) secreted by LPS- stimulated monocytes. Expression of COX-2 protein was partially reduced by treatment of anti-human GM-CSF Ab. Activation of STAT5 was inhibited by either IL-10 or dexamethasone (Dex), but not by aspirin. IL-10 blocked activation of STAT5 indirectly by suppressing GM-CSF production, while Dex inhibited this activation both directly and indirectly. Taken together, these results suggest that in addition to other transcription factors, STAT5 plays an important role in activation of monocytes by LPS, and that STAT5 is another target for IL-10 and Dex to inhibit COX-2 expression in activated monocytes..
69. Yosuke Tanaka, Hitoshi Nakashima, Takeshi Otsuka, Yoshiaki Nemoto, Hiroaki Niiro, Kunihiro Yamaoka, Ei Ichi Ogami, Yojiro Arinobu, Hidenori Tachida, Takashi Imamura, Yoshiyuki Niho, Detection of polymorphisms within the human IL10 receptor cDNA gene sequence by RT-PCR RFLP, Immunogenetics, 10.1007/s002510050301, 46, 5, 439-441, 1997.09.