イヌの同種腎移植における特異的免疫抑制に関する基礎的研究 : とくに抗Ia単一クローン性抗体による拒絶反応抑制効果について

臓器移植における最も重要な問題は移植後に発現する移植免疫反応すなわち拒絶反応をいかに制御するかにある。　1960年代に開発された移植免疫抑制法はそれまで不可能であった同種移植を可能とし，とくに腎移植においてはヒトにおける臨床応用を一般的なものとするに至っている。　しかしながら，現在の免疫抑制法はいまだ不完全なものであり，移植腎の機能を喪失する原因として拒絶反応が約80％を占めているといわれる。また，現在免疫抑制法の主流をなしている免疫抑制剤は，非特異的な免疫抑制法であるため，感染に対する抵抗性の低下ならびに肺炎．髄膜炎などの重篤な合併症の原因となり，さらには造血機能，骨代謝機能ならびに肝機能の障害や発癌率を増加するなどの副作用を発現している。　近年の著しい免疫遺伝学の進歩により，移植の拒絶反応は根本的には遺伝的に規制された免疫応答であり，なかでも免疫学的に自己を決定している移植抗原が拒絶反応の主因をなし，主要組織適合遺伝子複合体(major histocompatibility complex，以下，MHC)により最も強い移植抗原が支配されていることが明らかになっている。さらに，興味あることはどの動物種においてもMHCがそれぞれ1つずつ存在していることである。　現在MHCの研究が最も進んでいるのはマウスであり，マウスのMHCは第17染色体上に存在しH-2複合体と命名されている。H-2複合体はK，I，S，G，Dの5つの領域にわけられ，I領域はさらにI-A，B，J，E，Cの亜領域にわけられて，特定の抗原に対する免疫応答を支配している。Ia抗原はI-A，E，C亜領域に支配される3種類が明らかにされ，細胞表面上に存在している。とくにI-A亜領域に規定されるIa抗原が極めて強い移植抗原であり，免疫応答遺伝子と密接な関係にあるとされている。また，マウスのIa抗原に相当する分子がヒト，サル，モルモットならびにラットにおいても存在すること(Ia様抗原)が報告されている。　このように，臓器移植における免疫応答に最も強く関与して個体を表現し，認識するものがMHCであり，とくにI-A亜領域に規定されるIa抗原によって移植片が認識され拒絶反応が発現される。このため一般の免疫能に影響を及ぼさずに移植における拒絶反応のみを抑制する特異的な免疫抑制法の開発が切望されている。　今回著者は，腎移植における特異的免疫抑制法の基礎的研究として，免疫応答のなかで重要な役割をもつIa抗原の関与する特定の免疫応答，すなわち拒絶反応のみを人為的に制御することを目的とし，Ia抗原に特異的な単一クローン性抗体(monoclonal antibody，以下，MCA)，すなわち抗IaMCAを用いてイヌの同種腎移植を実施し，その拒絶反応抑制効果について検討した。　実験に使用した抗IaMCAは，H-2複合体のうち，I-A亜領域のKハプロタイプのみに特異的に反応するマウス由来の抗体である。　予備実験として，はじめにマウスのIa抗原に相当する抗原がイヌにおいても存在し，マウス由来の抗IaMCAと種を越えて交叉反応を示すか否かを検討した。すなわちイヌリンパ球とマウス由来の抗IaMCAとの交叉反応の有無をFITC(Fluorescein isothiocyanate)を結合したヤギ抗マウス抗体を用いて，染色陽性率をFACS(Fluorescent activated cell sorter)により測定し，検討した。その結果，Controlにおける染色陽性率は5.4％であったのに対し，抗IaMCAにより処理することによってその陽性率は21.9％と高値を示し，イヌリンパ球においてもマウスのIa抗原に相当するIa様抗原が存在し，マウス由来の抗IaMCAと交叉反応を示すことが確認された。　ついでイヌリンパ球において証明されたIa様抗原が，臓器あるいは組織においても同様に存在するか否かについて検討するためにマウス，ラットならびにイヌの腎臓を，抗IaMCAで処理し，FITC間接蛍光抗体法により腎臓内のIaまたはIa様抗原の検索を行った。その結果，マウスの腎臓におけるIa抗原の分布は糸球体およびその周囲に強い陽性反応を示し，他の部分ではほとんど陽性反応が見られなかった。また，ラットの腎臓においては糸球体に対する陽性反応は弱く，細～中動脈の内皮細胞およびその周囲に強い陽性反応を示したが，イヌの腎臓においては一様に糸球体，尿細管上皮細胞および動脈の内皮細胞に陽性反応を示した。したがって，マウス，ラットならびにイヌの腎組織においてもリンパ球と同様にIaまたはIa様抗体が存在することが確認されたが，その分布に関しては動物種により異なることが推測された。　以上のような予備実験において，マウス由来の抗IaMCAによるイヌの腎移植における拒絶反応抑制効果が示唆されたことより，イヌの同種腎移植実験を実施した。　実験は雑種成犬61例を4群にわけて行った。Group I(18例)は対照群として移植腎ならびに宿主に対し抗IaMCAを使用しなかった。Group II(25例)は摘出した移植腎を抗IaMCA10mlで灌流し，30分間4℃で反応させたのちに，移植を行った。Group III(8例)はGroup IIと同様の方法で移植腎を処理したのちに移植を行い，移植後宿主に対して抗IaMCA10mlを隔日に静脈内に投与した。Group IV(10例)は移植腎は対照群と同様無処置のまま移植し，移植後宿主に対して抗IaMCA10mlを隔日に静脈内に投与した。各群ともに移植後宿主の固有腎は摘出し，他の免疫抑制剤は一切使用しなかった。　その結果，Group Iにおける生存日数は5～13日，平均7.2±2.1日であった。剖検時の移植腎の病理組織学的検査では，全例典型的な急性拒絶反応の所見を示していた。　Group IIにおける生存日数は10～24日，平均14.4±3.6日であり，Group Iと比較し極めて有意(P<0.01)の差が認められた。移植腎の病理組織学的検査では，生存日数10～13日の4例においては，急性拒絶反応の所見に加え腎皮質における層状の高度の細胞浸潤が認められ，さらに髄質部における広範囲の壊死像を示すDIC(播発性血管内凝固．disseminated intravascular coagulation)様の合併症を呈する所見が観察された。また，20日以上正存した3例においては腎皮質の層状の変化はみられず，拒絶反応にDIC様の合併を呈している所見であった。その他の例については急性拒絶反応の所見のみを示した。　Group IIIにおける生存日数は7～14日，平均11.1±2.4日であり，Group Iと比較し有意(P<0.05)の差がみられた。移植腎の病理組織所見では，2例にGroup IIにおいても観察された腎皮質における層状の特徴的変化がみられた。その他の例においては全例急性拒絶反応の所見が観察された。　Group IVにおける生存日数は7～12日，平均9.4±2.1日であり，Group Iと比較し有意(P<0.05)の差がみられた。2例に抗IaMCA投与直後にアナフィラキシー様ショックがみられた。移植腎の組織にはほとんどの例で急性拒絶反応の所見が観察された。　以上の結果から，イヌの同種腎移植においてマウス由来の抗IaMCAを前もって移植腎に灌流，または移植後，宿主に静脈内投与あるいはその両者を併用することにより，明らかに術後の生存日数の延長が認められ，とくに移植腎のみを抗IaMCAで灌流したのちに移植する方法が最も生存日数を延長させる効果があった。また，抗IaMCAの移植腎灌流と術後静脈内投与の併用または術後静脈内投与のみの方法では，移植腎のみの灌流法ほど生存期間の延長効果は著明でなかったが，対照群よりは有意に生存日数が延長した。したがって，抗IaMCAを用いることにより，腎移植後の生存日数が延長する理由は，抗IaMCAが拒絶反応のtriggerとなる腎臓のIa様抗原をブロックし，宿主側のhelper T cellによる抗原認識が抑制されるために，拒絶反応が抑制されるのであろうと推測された。　このように，抗IaMCAを同種腎移植に用いることにより，明らかに移植後の生存日数は延長したが，移植腎の完全な生着例は得られず，病理組織学的検査にみられたように抗IaMCAを使用した群においてもほぼ全例において急性拒絶反応の所見が観察されたことから，現段階においては抗IaMCAにより，完全に拒絶反応を抑制するまでには至っていない。しかしながら，今回使用した抗IaMCAは，マウスのI-A亜領域のkハプロタイプに規定されるIa抗原のみと特異的に反応する抗体であり，I-A亜領域のk以外のハプロタイプまたはI-E，C亜領の各ハプロタイプとは反応しないため，それらに規定されるIa抗原により拒絶反応が出現したためであろうと考えられる。　今後，I-A亜領域のkハプロタイプ以外のIa抗原を規定するI-A，E，C亜領域の各ハプロタイプに対する抗IaMCAのほか，H-2複合体のK，D領域に規定される標的抗原など，移植免疫反応を司っている遺伝領域にそれぞれ反応する特異的なMCAを用いることによって，それらが関与する免疫反応のみを特異的に制御することができ，従来の非特異的な免疫抑制剤を使用することなく，同種腎移植を成功させることが可能となるであろう。　さらに異種の認識に関与する遺伝子領域の解明による種を超えた異種移植の可能性をも示唆するものと考えられる。One of the most important problems in the field of organ transplantation today is to find the safest and specific way of how to suppress the rejection reaction which must follow allogenic organ transplantation. In order to minimized the rejection reaction, the administration of nonspecific and potent immunosuppressive drugs have been currently used in many transplantation cases. However, we all know that drugs have serious drawbacks such as fatal infectious and other complication in the kidney transplantation patients, because those drugs is due to total suppression of immune reactions. One of the specific immunosuppression would be to block the antigen recognition system of immune response. In this line of thinking, author has developed a specific monoclonal antibody (anti Ia MCA) directed to mouse Ia antigen and used to the canine renal transplantation. The aim of this experiments are two folds; No.1 to find if anti Ia MCA is capable of cross reacting the dog lymphocytes and if Ia like antigen exists in the dog kidney and No.2; to find if anti Ia MCA is able to prolong the survival time of canine kidney allograft. Canine kidney transplantations have been performed in four experimental groups. Group I (18 cases) comprised of control. Group II (25 cases) was simple renal perfusion with anti Ia MCA. Group III (8 cases) was simple renal perfusion and intravenous injection with anti Ia MCA. Group IV (10 cases) was only intravenous injection with anti Ia MCA. All animals did\u27nt received postoperative immunosuppression. The results were sammarized as follows: When canine lymphocyte and kidney tissue were treated with anti Ia MCA followed, lymphocytes and kidney tissues were found to be possitive by FACS indirect fluorescent staining. This suggests a cross reaction between the Ia antigens of the mouse and the dog species. The experiments of canine renal allograft transplantation with anti Ia MCA indicated that the mouse anti Ia MCA can prolonge the survival time of canine renal allografts by simple organ perfusion. The dogs of group I survived from 5 to 13 days, mean survival time being 7 days. Group II survived from 10 to 24 days, mean 14 days. Group III survived from 7 to 14 days, mean 11 days. Group IV survived from 7 to 12 days, mean 9 days. The anti Ia MCA was appearently able to prolong the survival of the canine renal allografts.獣医学博士麻布大

Breadand Effervescent Beverage Productions with Local Microbes for the Local Revitalization

Local microbes such as yeasts fungi and bacilli, were isolated and used to apply for the food processing (bread making) and the production of an effervescent beverage (fruits kvass) for promotion of the local revitalization. Our yeast isolates could contribute to make the breads that can sell in alocal bakery shop.  Furthermore, the same yeasts could be used for making the fruit kvass (a Russian effervescent beverage which was fermented by yeast) with local fruit products (lemon, dry grape, mandarin orange and apple that were produced in Hiroshima prefecture in Japan). Development of merchandise having the local brand, which was made with only products of Hiroshima prefecture was attempted. Results of these activities demonstrate that the search of local microbes can help to establish the regional brand product related to its local area. They also showed that the local microbes have the potential ability to lead the local revitalization and the local brand product

Endoscopic and clinical evaluation of treatment and prognosis of Cronkhite-Canada syndrome: a Japanese nationwide survey.

BackgroundFirst reported in 1955, Cronkhite-Canada syndrome (CCS), a rare syndrome characterized by ectodermal abnormalities and inflammatory changes of the gastrointestinal tract mucosa, has been associated with a poor prognosis and life-threatening malignant complications. In a large population survey, we endeavored to characterize the course and treatment outcome of CCS through clinical and endoscopic assessment, and to explore its optimal treatment and surveillance strategy.MethodsA retrospective analysis of 210 patients with CCS was conducted via a questionnaire-based nationwide survey of 983 teaching hospitals located throughout Japan. We assessed clinical features, endoscopic findings, treatments used, and short- and long-term outcomes.ResultsThe average age at diagnosis was 63.5 years. In all cases, upper or lower gastrointestinal tract polyposis was confirmed, accompanied by characteristic ectodermal abnormalities. Of the treatments used, oral corticosteroids (30-49 mg/day) were the most effective treatment for active disease, with adjunctive nutritional support considered beneficial. With corticosteroid treatment, abdominal symptoms were relieved within a few months, whereas polyp regression often required more than 6 months. Maintenance of endoscopic remission with or without steroids for 3 years significantly lowered the development of CCS-related cancer, compared with relapsers or nonresponders, underscoring the importance of sustained endoscopic remission for cancer prevention.ConclusionsThe prognosis of CCS has greatly improved through the use of improved medical treatment. Although CCS continues to be relentlessly progressive, carrying a high cancer risk, a sufficient dose and duration of corticosteroid therapy accompanied by nutritional support and periodic endoscopic surveillance appears to improve its natural history

Multiple Regression Analysis for Grading and Prognosis of Cubital Tunnel Syndrome:Assessment of Akahori’s Classification

The purpose of this study was to quantitatively evaluate Akahori's preoperative classification of cubital tunnel syndrome. We analyzed the results for 57 elbows that were treated by a simple decompression procedure from 1997 to 2004. The relationship between each item of Akahori's preoperative classification and clinical stage was investigated based on the parameter distribution. We evaluated Akahori's classification system using multiple regression analysis, and investigated the association between the stage and treatment results. The usefulness of the regression equation was evaluated by analysis of variance of the expected and observed scores. In the parameter distribution, each item of Akahori's classification was mostly associated with the stage, but it was difficult to judge the severity of palsy. In the mathematical evaluation, the most effective item in determining the stage was sensory conduction velocity. It was demonstrated that the established regression equation was highly reliable (R＝0.922). Akahori's preoperative classification can also be used in postoperative classification, and this classification was correlated with postoperative prognosis. Our results indicate that Akahori's preoperative classification is a suitable system. It is reliable, reproducible and well-correlated with the postoperative prognosis. In addition, the established prediction formula is useful to reduce the diagnostic complexity of Akahori's classification

Sodium valproate blocks the transforming growth factor (TGF)-β1 autocrine loop and attenuates the TGF-β1-induced collagen synthesis in a human hepatic stellate cell line

13301甲第3918号博士（医学）金沢大学博士論文要旨Abstract 以下に掲載：International Journal of Molecular Medicine 28(6) pp.919-925 2011.Spandidos Publicatons. 共著者：Toshifumi Watanabe, Hidehiro Tajima, Hayashi Hironori, Hisatoshi Nakagawara, Ichiro Ohinishi, Hiroyuki Takamura, Itasu Ninomiya, Hirohisa Kitagawa, Sachio Fushida, Takashi Tani, Takashi Fujimura, Tetsuo Ota, Tomohiko Wakayama, Shoichi Iseki, Shinichi Harad

統合失調症初発エピソードの認知機能障害の経過と発症5 年経過群との比較

OBJECTIVE: The course of neurocognitive deficits in schizophrenia has not yet been established. Therefore, we followed patients with first-episode schizophrenia to verify the course of these deficits. METHODS:In Study 1, tests of neurocognitive functioning were administered to patients with first-episode schizophrenia (FE group) every 6 months. Of the 26 patients who completed the baseline assessment, 19 completed a 6-month follow-up, and 13 completed a 1-year follow-up. In Study 2, 19 patients in FE group at 6-months when the neuropsychological measures was less influenced by psychotic symptoms and other patients who experienced schizophrenia 5-years earlier (5-year group) were compared. RESULTS:In Study 1, verbal memory, motor speed, and executive functions significantly improved at the 1-year follow-up. In Study 2, patients in 5-year group performed worse in verbal memory and executive functions than patients in FE at 6-month group, but marginally but significantly better in verbal fluency. CONCLUSIONS:Verbal memory, executive functions, and verbal fluency were significantly different between 5-year group and FE at 6-month group, and may indicate progression of schizophrenia. Executive functions may reflect the state of psychosis. Working memory and processing speed which did not change significantly from onset are needed to verify the course in further research.博士（医学）・甲610号・平成26年3月17

Bone regeneration of induced pluripotent stem cells derived from peripheral blood cells in collagen sponge scaffolds

Stem cell-based regeneration therapy offers new therapeutic options for patients with bone defects because of significant advances in stem cell research. Although bone marrow mesenchymal stem cells are the ideal material for bone regeneration therapy using stem cell, they are difficult to obtain. Induced pluripotent stem cells (iPSCs) are now considered an attractive tool in bone tissue engineering. Recently, the efficiency of establishing iPSCs has been improved by the use of the Sendai virus vector, and it has become easier to establish iPSCs from several type of somatic cells. In our previous study, we reported a method to purify osteogenic cells from iPSCs. Objective: This study aimed to evaluate the osteogenic ability of iPSCs derived from peripheral blood cells. Methodology: Mononuclear cells (MNCs) were obtained from human peripheral blood. Subsequently, T cells were selectively obtained from these MNCs and iPSCs were established using Sendai virus vectors. Established iPSCs were evaluated by the expression of undifferentiated markers and teratoma formation assays. Osteoblasts were induced from these iPSCs and evaluated by the expression of osteoblast markers. Additionally, the induced osteoblasts were transplanted into rat critical size calvaria bone defect models with collagen sponge scaffolds. Samples were evaluated by radiographical and histological assessments. Results: Induced osteoblasts expressed several osteoblast-specific markers. The results of radiographical and histological assessments revealed that the cell transplant group had bone formations superior to those of the control group.&nbsp;Conclusions: This study suggests that peripheral blood MNCs have the potential to differentiate into osteoblasts. Although there are some hurdles in iPSC transplantation, osteoblasts obtained from MNC-iPSCs could be applied to bone regeneration therapy in the future

G-band and Hard X-ray Emissions of the 2006 December 14 flare observed by Hinode/SOT and RHESSI

We report on G-band emission observed by the Solar Optical Telescope onboard the Hinode satellite in association with the X1.5-class flare on 2006 December 14. The G-band enhancements originate from the footpoints of flaring coronal magnetic loops, coinciding with non-thermal hard X-ray bremsstrahlung sources observed by the Reuven Ramaty High Energy Solar Spectroscopic Imager. At the available 2 minute cadence, the G-band and hard X-ray intensities are furthermore well correlated in time. Assuming that the G-band enhancements are continuum emission from a blackbody, we derived the total radiative losses of the white-light flare (white-light power). If the G-band enhancements additionally have a contribution from lines, the derived values are overestimates. We compare the white-light power with the power in hard X-ray producing electrons using the thick target assumption. Independent of the cutoff energy of the accelerated electron spectrum, the white-light power and the power of accelerated electrons are roughly proportional. Using the observed upper limit of ~30 keV for the cutoff energy, the hard X-ray producing electrons provide at least a factor of 2 more power than needed to produce the white-light emission. For electrons above 40 keV, the powers roughly match for all four of the time intervals available during the impulsive phase. Hence, the flare-accelerated electrons contain enough energy to produce the white-light flare emissions. The observed correlation in time, space, and power strongly suggests that electron acceleration and white-light production in solar flares are closely related. However, the results also call attention to the inconsistency in apparent source heights of the hard X-ray (chromosphere) and white-light (upper photosphere) sources.Comment: 15 pages, 7 figures, accepted for publication in Ap

Amino acid residues in the transmembrane domain of the type 1 sigma receptor critical for ligand binding

AbstractThe type 1 sigma receptor expressed in Xenopus oocytes showed binding abilities for the sigma-1 ligands, [3H](+)pentazocine and [3H]NE-100, with similar kinetic properties as observed in native tissue membranes. Amino acid substitutions (Ser99Ala, Tyr103Phe and di-Leu105,106di-Ala) in the transmembrane domain did not alter the expression levels of the type 1 sigma receptor as determined by immunoblot analysis using an anti-type 1 sigma receptor antiserum. By contrast, ligand binding was significantly suppressed by the substitutions. These findings provide evidence that the transmembrane domain of the type 1 sigma receptor plays a critical role in ligand binding of this receptor