伊藤 弘康

Indoleamine 2,3-dioxygenase exerts intense immunomodulatory effects due to enzymatic activities that catalyze the breakdown of the essential amino acid L-tryptophan. The activity of indoleamine 2,3-dioxygenase can be estimated by measuring serum L-kynurenine concentrations. Here, we aimed to determine the role of L-kynurenine as a prognostic factor for peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) in a retrospective analysis of data derived from 31 consecutive patients between June 2000 and March 2013 who were histologically diagnosed with PTCL-NOS according to the World Health Organization classification and treated with 6 - 8 cycles of cyclophosphamide, doxorubicin or pirarubicin, vincristine, and prednisolone. L-kynurenine concentrations in serum samples collected at admission were measured using high-performance liquid chromatography. The median serum concentration of L-kynurenine was 3.28 (range 0.92-8.16) mu M. The L-kynurenine cutoff was set at 3.07 mu M using receiver operating characteristics curves. The complete remission rates of patients with L-kynurenine <3.07 and >= 3.07 mu M were 69% and 51%, respectively. The 5-year overall survival (OS) rates for patients with L-kynurenine < 3.07 and >= 3.07 mu M were 80.2% and 23.4%, respectively (p<0.001). More advanced age, poor performance status, elevated lactate dehydrogenase, an unfavorable International Prognostic Index, and a poor prognostic index for T-cell lymphoma were significantly worse factors for OS. Multivariate analyses revealed only L-kynurenine as an independent prognostic factor for OS. In conclusion, serum concentrations of L-kynurenine might comprise a novel prognostic factor with which to determine the outcomes of treatment for PTCL-NOS. Copyright (c) 2016 John Wiley & Sons, Ltd.

今回、筆者らはシスメックス社のCS-2400で測定可能となった血小板凝集能検査について評価する機会を得て、基礎検討を行った。CS-2400の同時再現性、オンボード安定性、共存物質の影響、いずれの検討結果も良好で、既存装置との相関性は、r=0.75〜0.84と良好であった。これまでは、専用装置が必要であった。血小板凝集能測定がルーチンと同じ全自動測定装置であるCS-2400で測定可能となり、作業工程の省力化や検査ワークフローの改善に寄与すると考えられる。(著者抄録)

化学発光酵素免疫測定法による肝臓型脂肪酸結合蛋白(L-FABP)測定試薬の基礎的性能評価を行った。再現性、希釈直線性、測定感度は良好な成績であり、共存物質における影響も認められなかった。また、従来法との相関性も良好であった。このことから、本測定試薬は日常検査における十分な基本性能を有していると考えられる。さらに臨床性能評価として、試験紙尿蛋白陰性患者においてCKD重症度分類リスク群別に尿中L-FABP/クレアチニン値の比較を行った。その結果、重症度リスクの増大に伴う尿中L-FABPの増加がみられたことから、尿蛋白陰性患者のような早期腎機能異常群における尿中L-FABPの有用性が期待される。(著者抄録)

Inflammatory response is required to proceed the optimal liver regeneration after liver injury. Recent reports demonstrated that inflammasomes are involved in the innate immune response. Several NOD-Like receptors (NLRs) participated in the formation of the inflammasomes. NACHT, LRR, and PYD domain-containing protein 3 (NALP3) belongs to the NLR families and recognizes adenosine triphosphate (ATP), crystals, and reactive oxygen species. The present study examined the effect of inflammasomes on the process of liver regeneration using NALP3 knockout (KO) mice. The activation of inflammasomes in the liver was induced after 70% partial hepatectomy (PHx). The liver-to-body weight ratio was significantly decreased in NALP3-KO mice compared to that in WT mice after PHx. The number of Ki67-positive cells and the expression of Cyclin D1 and E1 after PHx were reduced in NALP3-KO mice compared to WT mice. The expression of pro-inflammatory cytokines (IL-1 beta, TNF-alpha, and IL-6) were decreased in the remnant liver of NALP3-KO mice compared to WT mice. Flow cytometric analysis revealed that the expression of chemokines was decreased and the accumulation of CD11b-positive cells was suppressed in NALP3-KO mice after PHx. The treatment with ATP, which is a ligand to NALP3, increased the liver-to-body weight ratio in WT mice. These results indicate that NALP3 signaling is required for the induction of inflammatory response and the development of liver regeneration after PHx.

The inflammatory response accelerates early liver regeneration after liver injury and resection. Recent studies have demonstrated that indoleamine 2,3-dioxygenase-1 (IDO1) suppresses the activation of inflammatory cells and induces immune tolerance. In this study, we examined the role of IDO1 in liver regeneration after partial hepatectomy (PHx). WT or IDO1-knockout (IDO1-KO) mice received 70% PHx. The liver-body weight ratio after PHx was measured and hepatocyte growth was assessed by immunostaining. The expression of cell cycle genes and pro-inflammatory cytokines in the liver was analyzed by quantitative RT-PCR. In addition, 1-methyl-dl-tryptophan (1-MT), which is an IDO1 inhibitory agent, was given to WT mice and the liver-body weight ratio was measured after PHx. The liver-body weight ratio was significantly increased in IDO1-KO mice compared with that in WT mice after PHx. More Ki-67-positive cells were present in IDO1-KO mice than in WT mice after PHx. The expression of cell cycle genes (cyclin D1, cyclin E) and pro-inflammatory cytokines (IL-1 beta, TNF-alpha and IL-6) was up-regulated in the remnant liver of IDO1-KO mice compared with WT mice. Moreover, treatment with 1-MT promoted liver regeneration. IDO1 deficiency promoted early liver regeneration after PHx, indicating that IDO1 suppresses the production of inflammatory cytokines and subsequently inhibits hepatocyte proliferation during liver regeneration.

ObjectivesThe aim of this study was to evaluate the relationships among aging, muscle strength, and image feature analysis of the quadriceps femoris muscle and to evaluate the relationship between aging, muscle strength, and sonographic findings. MethodsOne hundred forty-five healthy volunteers participated in the study. The participants were classified into 6 groups on the basis of sex and age. To assess muscle quality, texture analysis was defined with the following parameters: mean, skewness, kurtosis, inverse difference moment, sum of entropy, and angular second moment. The knee extension force in the sitting position and thickness of the quadriceps femoris muscle were also measured. ResultsThe quadriceps femoris thickness, skewness, kurtosis, inverse difference moment, angular second moment, and muscle strength were significantly decreased in elderly participants versus those in the younger and middle-aged groups (P < .05). In contrast, the mean and sum of entropy were significantly decreased in the younger group compared with the middle-aged and elderly groups. ConclusionsSonography has the capacity to quantitatively assess muscular morphologic changes due to aging and could be a valuable tool for early detection of musculoskeletal disorders.

下腿筋厚と筋内部エコーについて評価し、加齢に伴う変化や深部静脈血栓症との関連性について検討した。また、CT検査による骨格筋量評価を用いたサルコペニア診断基準と深部静脈血栓症陽性率についても検討した。その結果、age matching後の対象群において深部静脈血栓症の有無による比較検討を行い、深部静脈血栓症陽性群における有意な筋厚の減少とエコー輝度の上昇を認めた。また、サルコペニアの有無と関連する因子を明らかにするために解析を行った結果、筋線維性状がサルコペニアのの独立予測因子であり、エコーによる筋線維性状評価は筋質の低下を捉えられる可能性があった。

It remains unclearwhat cells are proper for the generation of induced pluripotent stem cells (iPSCs). Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5) is well known as a tissue stem cell and progenitor marker, both of which are reported to be sensitive to reprogramming. In the present study, we examined the reprogramming behavior of Lgr5-expressing cells (Lgr5+ cells). First, we compared reprogramming behavior using mouse Lgr5+ and Lgr5 negative (Lgr5-) hair follicles (HFs). The number of alkaline phosphatase staining-positive cells was lesser in a well of Lgr5+ HFs than in Lgr5-HFs; however, the ratio of Nanog+ SSEA1+ cells in the cell mixture derived from Lgr5+ HFs was much higher than that from Lgr5-HFs. Lgr5+ cells could be induced from mouse embryonic fibroblasts (MEFs) after transduction with Yamanaka factors. As shown in HFs, the progeny of Lgr5+ cells arising from MEFs highly converted into Nanog+ cells and did not form Nanog-colonies. The progeny represented the status of the late reprogramming phase to a higher degree than the nonprogeny. We also confirmed this using human Lg5+ cells. Our findings suggest that the use of Lgr5+ cells will minimize sorting efforts for obtaining superior iPSCs. (C) 2017 The Authors. Published by Elsevier B.V.

A persistent hepatitis B virus (HBV) infection is characterized by a lack of or a weak immune response to HBV, which may be reflective of tolerance to HBV. Efficient induction of HBV-specific immune response leads to the clearance of HBV in patients with a chronic HBV infection. CpG oligodeoxynucleotides (ODN) has a powerful adjuvant effect in HBV vaccination. A recent report demonstrated that the immunization by B/K CpG ODN (K3) wrapped by the nonagonistic Dectin-1 ligand, schizophyllan (SPG), namely K3-SPG, was more effective in the induction of antigen-specific immune response than that by K3. In this study, we examined the efficacy of K3-SPG as a HBV vaccine adjuvant. Wild-type (WT) mice and HBV transgenic (HBV-Tg) mice were subcutaneously immunized with hepatitis B surface antigen (HBsAg) alone, HBsAg and K3, or HBsAg and K3-SPG. The vaccination with HBsAg and K3-SPG significantly enhanced humoral and cellular immune response to HBV antigen compared to the other vaccinations in WT and HBV-Tg mice. K3-SPG induced the accumulation of dendritic cells (DCs) into draining lymph node and the activation of DCs. The expression of cytokines and chemokines related to Th1 and Th2 responses was upregulated after the vaccination including with K3-SPG. In conclusion, these results indicated that the vaccination using K3-SPG may overcome tolerance even in patients with chronic HBV infection.

Viral infections can give rise to secondary bacterial infections. In the present study, we examined the role of invariant natural killer T (iNKT) cells in lipopolysaccharide (LPS)-induced lethal shock during encephalomyocarditis virus (EMCV) infection. Wild-type (WT) mice and J alpha 18 gene knockout (1 alpha 18 KO) mice were inoculated with EMCV, 5 days prior to challenging with LPS. The survival rate of J alpha 18 KO mice subjected to EMCV and LPS was significantly higher than that of WT mice. TNF-alpha and nitric oxide (NO) production were increased in WT mice, than that in J alpha 18 KO mice, after the administration of EMCV and LPS. EMCV infection increased the number of iNKT cells and IFN-gamma production by iNKT cells in WT mice. Moreover, EMCV infection enhanced the expression of Toll-like receptor 4 (TLR4) in the lung and spleen. IFN-gamma also increased the expression of TLR4 in splenocytes. These findings indicated that EMCV infection activated iNKT cells, and IFN-gamma secreted from the iNKT cells up-regulated the expression of TLR4 in various tissues. As a result, EMCV-infected mice were susceptible to LPS and easily developed the lethal shock. In conclusion, iNKT cells were involved in the development of LPS-induced lethal shock during EMCV infection. (C) 2016 Elsevier GmbH. All rights reserved.

Immune cells constitute a large fraction of the tumor microenvironment and modulate tumor progression. Clinical data indicate that chronic inflammation is present at tumor sites and that IL4 in particular is upregulated. Here, we demonstrate that T follicular helper (Tfh) cells arise in tumor-draining lymph nodes where they produce an abundance of IL4. Deletion of IL4-expressing Tfh cells improves antitumor immunity, delays tumor growth, and reduces the generation of immunosuppressive myeloid cells in the lymph nodes. These findings suggest that IL4 from Tfh cells affects antitumor immunity and constitutes an attractive therapeutic target to reduce immunosuppression in the tumor microenvironment, and thus enhance the efficacy of cancer immunotherapy. Cancer Immunol Res; (C) 2016 AACR.

In the present study, we examined the role of indoleamine 2,3-dioxygenase (IDO) in the development of CCl4-induced hepatic fibrosis. The liver fibrosis induced by repetitive administration with CCl4 was aggravated in IDO-KO mice compared to WT mice. In IDO-KO mice treated with CCl4, the number of several inflammatory cells and the expression of pro-inflammatory cytokines increased in the liver. In the results, activated hepatic stellate cells (HSCs) and fibrogenic factors on HSCs increased after repetitive CCl4 administration in IDO-KO mice compared to WT mice. Moreover, the treatment with L-tryptophan aggravated the CCl4-induced hepatic fibrosis in WT mice. Our findings demonstrated that the IDO deficiency enhanced the inflammation in the liver and aggravated liver fibrosis in repetitive CCl4-treated mice.

新規肝線維化マーカ「Mac-2結合蛋白糖鎖修飾異性体(M2BPGi)」測定試薬の基礎的検討および臨床的性能について検討した。HCV感染症患者45例を対象とした。共存物質では、直接型ビリルビン21.0mg/dL、間接型ビリルビン21.0mg/dL、溶血ヘモグロビン510mg/dL、乳び1.490FTU、リウマトイド因子500IU/mLまで影響を受けなかった。低濃度及び高濃度検体ともに原点を通る良好な直線性を確認した。理論値で0.0014COIまでの最小検出感度を確認した。患者血清は4℃、-30℃の保存で安定した結果が得られた。肝線維化進行群における診断能は、AUC 0.92、感度85%、特異度94%で、高度肝線維化進行群では、AUC 0.83、感度83%、特異度75%で、従来の肝線維化関連マーカとほぼ同等の成績であった。

はじめに：本研究の目的は，超音波検査による棘下筋の脂肪浸潤の評価を行い，MRI結果や腱板断裂程度との関連性について検討することである．<br>対象と方法：腱板評価のためにMRIと超音波検査を施行した32例32肩（平均年齢68.2±11.4歳，男性20例，女性12例）を対象とした．棘下筋の脂肪浸潤については，MRIによるGoutallier分類に従い，stage0と1を軽度，stage2を中等度，stage3と4を併せて高度とした．超音波検査については，装置に搭載されているHistogram法を用い，皮下脂肪層，棘下筋内それぞれ3カ所より計測を行い，ROI中の平均階級値（MN）の平均値を指標とし，皮下脂肪を筋のMNで除した値をecho intensity ratio: ERとした．<br>結果：腱板断裂サイズと年齢の関連性については，断裂なし群が広範囲断裂群に比し有意に低下した．MRIにおける棘下筋脂肪浸潤評価とUS Histogram法の関連性では，皮下脂肪のMNは軽度が高度に比し有意に高値を示し，棘下筋のMNについては，軽度が高度に比し有意に低値を示した．ERについては，高度が軽度，中等度に比べ有意に高値を示した．<br>結語：今回の検討において超音波Histogram法は棘下筋の脂肪浸潤を簡便に定量評価できる有用な検査法であると考えられた．

Subcutaneous masses caused by foreign bodies are frequently encountered in daily practice. Although the majority of foreign bodies such as metals can be detected by radiography, substances such as vegetative materials or wood are difficult to detect. To our knowledge, only a few studies have described the sonographic characteristics of foreign bodies. Herein, we report 3 cases where we studied the sonographic characteristics of the foreign bodies in the dermis and subcutaneous tissue. Our results revealed the following 3 foreign bodies: (1) glass, (2) vegetative material, and (3) a pencil core. Thus, sonographic examination is useful for the detection of foreign bodies.

Alpha-garactosylceramide (GalCer) has been shown to have anti-tumor effect in the basic research and clinical studies. However, anti-tumor effect of GalCer is limited. The administration of GalCer increases the production of IFN-gamma which is involved in the suppression of tumor growth. On the other hand, the enhancement of IFN-gamma production increases immunosuppressive factors such as nitric oxide. This suppressive action might impair the anti-tumor effect of GalCer. In the present study, we evaluated the anti-tumor effect of GalCer in the absence of inducible nitric oxide synthase (iNOS). In lung metastatic model, the number of tumor nodules in the lung of iNOS-KO mice treated with GalCer was significantly reduced compared with that of WT mice treated with GalCer. Moreover, L-NAME, which is the inhibitor for iNOS, enhanced the anti-tumor effect of GalCer in lung metastatic model. The frequency of CD8+ cells in bronchoalveolar lavage fluid increased in iNOS-KO mice treated with GalCer. The administration of GalCer increased the frequency of myeloidderived suppressor cells (MDSCs) in the lung from tumor-bearing WT mice, but the increase of MDSCs in the lung was not induced in iNOS-KO mice. The subcutaneous tumor experiments revealed that the administration of GalCer in the absence of iNOS expression significantly enhanced the induction of tumor antigen-specific response. Finally, our results indicated that the inhibition of iNOS expression could enhance the therapeutic efficacy of GalCer via the increase of tumor antigen-specific immune response and the suppression of MDSCs.

Skin wound healing is a complex process involving several stages that include inflammation, proliferation, and remodeling. In the inflammatory phase, pro-inflammatory cytokines and chemokines are induced at the wound site and, they contribute to the development of wound healing. These cytokines also induce indoleamine 2,3-dioxygenase (IDO1) activity; this is the rate-limiting and first enzyme in the L-tryptophan (TRP)-L-kynurenine (KYN) pathway. This study examined the effect of IDO1 on the process of skin wound healing. The expression of the Idol mRNA was enhanced after creating a wound in wildtype (WT) mice. TRP concentration was simultaneously reduced at the wound site. The rate of wound healing in IDO1 knockout (IDO-KO) mice was significantly higher than that in WT mice. 1-Methyl-DL-tryptophan (1-MT), a potent inhibitor of IDO1, increased the rate of wound healing in WT mice. The administration of TRP accelerated wound healing in vivo and in an in vitro experimental model, whereas the rate of wound healing was not affected by the administration of KYN. The present study identifies the role of IDO1 in skin wound healing, and indicates that the local administration of 1-MT or TRP may provide an effective strategy for accelerating wound healing. (C) 2015 Elsevier Ltd. All rights reserved.

The immunomodulatory effects of indoleamine 2,3-dioxygenase (IDO) are ascribed to its ability to catalyze breakdown of the essential amino acid L-tryptophan. We applied reverse transcription-polymerase chain reaction (RT-PCR) to examine IDO mRNA expression in acute myeloid leukemia (AML) blasts, and investigated its clinical significance. We enrolled 62 patients with AML between April 2005 and March 2013. Bone marrow-derived mononuclear fractions were separated and extracted mRNA was amplified by PCR. RT-PCR showed that the bone marrow of 23 patients expressed IDO mRNA but not in 39. IDO mRNA expression did not significantly differ among cytogenetic risk profiles. The 3-year overall survival rates for patients with and without IDO mRNA expression were 39% and 74%, respectively (p &lt; 0.005). The rates for patients with intermediate-risk cytogenetics with and without IDO mRNA expression were 16% and 70%, respectively (p &lt; 0.005). The expression of IDO mRNA was associated with a poor prognosis of AML.

Toll-like receptor (TLR) agonists have been shown to have anti-tumour activity in basic research and clinical studies. However, TLR agonist monotherapy does not sufficiently eliminate tumours. Activation of the innate immune response by TLR agonists is effective at driving adaptive immunity via interleukin-12 (IL-12) or IL-1, but is counteracted by the simultaneous induction of immunosuppressive cytokines and other molecules, including IL-10, transforming growth factor-, and indoleamine 2,3-dioxygenase (IDO). In the present study, we evaluated the anti-cancer effect of the TLR7 agonist, imiquimod (IMQ), in the absence of IDO activity. The administration of IMQ in IDO knockout (KO) mice inoculated with tumour cells significantly suppressed tumour progression compared with that in wild-type (WT) mice, and improved the survival rate. Moreover, injection with IMQ enhanced the tumour antigen-specific T helper type 1 response in IDO-KO mice with tumours. Combination therapy with IMQ and an IDO inhibitor also significantly inhibited tumour growth. Our results indicated that the enhancement of IDO expression with TLR agonists in cancer treatment might impair host anti-tumour immunity while the inhibition of IDO could enhance the therapeutic efficacy of TLR agonists via the increase of T helper type 1 immune response.

Toll-like receptor (TLR) agonists have been shown to have anti-tumor activity in basic research and clinical studies. However, TLR agonist monotherapy in cancer treatment dose not sufficiently eliminate tumors. Activation of the innate immune response by TLR agonists and other pathogen-associated molecular patterns is effective for driving adaptive immunity via interleukin (IL)-12 or IL-1, but is counteracted by the simultaneous induction of immunosuppressive cytokines and other molecules, including IL-10, tumor growth factor-beta, and induced nitric oxide synthase (iNOS). In the present study, we evaluated the anticancer effect of the TLR7 agonist, imiquimod (IMQ), in the absence of iNOS. The administration of IMQ in iNOS-knockout (KO) mice implanted with tumor cells significantly suppressed tumor progression as compared to that in wild-type mice and improved the survival rate. Moreover, injection with IMQ enhanced the tumor antigen-specific Th1 response in iNOS-KO mice with tumors. The enhancement of the antigen-specific Th1 response was associated with an increase in IL-2 and IL-12b expressions in the tumor-draining lymph nodes. Combination therapy with IMQ and an iNOS inhibitor also significantly inhibited tumor growth in the established tumor model. Finally, our results indicated that the enhancement of iNOS expression through the administration with TLR agonists impairs host anti-tumor immunity, while the inhibition of iNOS could enhance the therapeutic efficacy of TLR agonists via the increase in Th1 immune response.

Purposes: The purposes of this study were to assess blood flow in the shoulders of patients with rotator cuff tears using the Doppler ultrasonography (US) and to evaluate the relationship between blood flow and tear size.<br> Methods: From August 2010 to August 2013, 87 patients with rotator cuff tears （RCTs）and 20 healthy volunteers (controls) were enrolled in this study. The patients were divided into two groups depending on the presence or absence of night pain. The peak systolic velocity (PSV) and resistance index (RI) of the anterior humeral circumflex artery (AHCA) were evaluated using pulse Doppler US. The PSV and RI were compared between the aflected and unaflected sides in patients and between the dominant and nondominant sides in controls.<br> Results: No significant difference in PSV and RI was noted between the dominant and nondominant sides in the controls. In the patients with RCTs, PSV of the AHCA in the affected side was significantly higher than that in the unaffected side (p 〈 0.001). PSV of the AHCA in the group with night pain was significantly higher than that in the group without night pain (p 〈 0.001). Moreover, the patients with night pain showed significantly lower RI on the affected side than on the unaffected side (p＝0.012). Meanwhile, no significant relationships were observed between tear size and blood flow of the AHCA. The receiver operating characteristic curve of the PSV revealed that the area under the curve was 0.85 with an optimal cutoff value of 20.5 cm/s, yielding 83% sensitivity and 86% specificity.<br> Conclusion: These results suggest that assessment of blood flow with Doppler US is useful for the diagnosis of RCT, especially in patients with night pain.

© 2015, Springer International Publishing Switzerland. The kynurenine (KYN) pathway is the major route of L-tryptophan (L-TRP) catabolism and an anabolic source of nicotinamide-containing nucleotide. To date, three enzymes that catalyze the first and rate-limiting step in the KYN pathway of TRP metabolism have been described: indoleamine 2,3-dioxygenase (IDO) 1, IDO2, and L-tryptophan 2,3-dioxygenase (TDO). In this chapter, we focus on the role of IDO1 in various infectious diseases. IDO1 has a much broader substrate profile for indoleamine-containing compounds and is induced by several proinflammatory cytokines. Substantial increases in the TRP-KYN pathway metabolites occur in human brain, blood, and systemic tissues during immune activation. This enzyme also plays a key role in the immunomodulatory effects on several types of immune cells. Originally known for its regulatory function during pregnancy and chronic inflammation in tumorigenesis, the activity of IDO1 seems to modify the inflammatory state of infectious diseases. Understanding the regulation of IDO1 and the subsequent biochemical reactions is essential for the design of therapeutic strategies in certain immune diseases. Therefore, we will discuss current knowledge about the role of IDO1 and its metabolites during various infectious diseases, e.g., infection by hepatitis virus, HIV, influenza virus, encephalomyocarditis virus, and parasites. Especially the regulation of type I interferon (IFN) production via IDO1 in these infectious diseases is discussed.