脳脊髄液の性状が脳動脈壁組織脆弱性に与える影響についての検討

金沢大学医薬保健研究域医学系法医解剖事例における小型の未破裂脳動脈瘤の頻度と分布を、脳脊髄液(CSF)の生化学的性状と併せて解析した結果、CSF中のゼラチナーゼ(galatinase A及びgalatinase B)活性が脳動脈瘤早期病変の鋭敏な指標となりうる（p<0.001）事が分かった。また、CSF中のゼラチン分解活性は、局所におけるCa調節を司るα-Klotho蛋白の部位別測定値（脳槽CSF、側脳室CSF及び血清中に各々542～5,461 pg/mL、2,758～5,971 pg/mL及び600～5,556 pg/mL）とも連動しており、病理組織学的解析と併せてCa濃度依存性の脳動脈瘤進展のメカニズムが示唆された。The frequency and distribution of unruptured cerebral aneurysms of minimal size in forensic autopsy cases were analyzed, measuring the activities of galatinases in cerebrospinal fluid(CSF). The results showed that gelatinolytic activities of CSF generated from gelatinase A and B can serve as good markers of early-stage cerebral aneurysm（p<0.001）. Moreover the gelatinolytic activities of CSF demonstrated parallels with the concentration of α-Klotho protein by which Ca deposition is controlled locally in the body. Therefore, combined with the microscopic findings, these Ca-dependent biochemical values of CSF can help to predict the development of cerebral aneurysms.研究課題/領域番号:26460874, 研究期間(年度):2014-04-01 - 2017-03-3

ヒト血管壁における組織脆弱性評価の研究

平成23～25年度に行った100例以上の法医解剖事例について、大動脈、頸動脈、脳動脈さらに脈絡叢の組織材料、血清及び脳脊髄液のゼラチナーゼ分解活性の検討を行った。身元不明で開始して後に身元が判明した司法解剖事例から、大動脈及び冠動脈のびまん性内膜肥厚及び組織内ゼラチン分解活性が年齢推定に有効であることが分かった。血管壁組織脆弱性の視点から、血管病変の危険因子である高血圧をもたらす、未検査の続発性アルドステロン症の頻度を組織学的に調べ、充分注意が払われていないことに対して警鐘を鳴らした。動物実験では、血管のびまん性内膜肥厚は線維化の一種であるという考えから自己免疫疾患モデルマウスの解析を行った。With regard to more than 100 cases subjected to forensic autopsy between 2011-2013, gelatinase degrading activity was investigated in aorta, carotid and cerebral arteries and choroid plexus tissue samples in addition to serum and cerebrospinal fluid. In forensic autopsies of initially unidentified cases in which subsequently identity was established, aorta and coronary artery diffuse intimal thickening and tissue gelatinolytic activity were found to be useful in the determination of age. From the viewpoint of vascular wall tissue fragility, we histologically investigated the frequency of unexamined secondary aldosteronism, a cause of hypertension and recognized risk factor for vascular lesions, and based on our findings raised an alarm that sufficient attention is not being paid to this important issue. In animal experiments, based on the contention that vascular diffuse intimal thickening represents one type of fibrosis, analysis using an autoimmune disease model mouse was conducted.研究課題/領域番号:23590844, 研究期間(年度):2011-201

血管壁の極性とプロテアーゼ活性に着目した頭蓋内組織脆弱性の解析

金沢大学医薬保健研究域医学系The frequency and distribution of unruptured cerebral small aneurysms and aneurysms of minimal size in forensic autopsy cases were analyzed, measuring the activities of galatinases in cerebrospinal fluid(CSF). However the results showed that gelatinolytic activities of CSF generated from gelatinase A and B can serve as good markers of early-stage cerebral aneurysm(p<0.001), the gelatinolytic activities of CSF did not demonstrate parallels enough with the concentration of α-klotho protein by which Ca deposition is controlled locally in the body. That means these Ca-dependent biochemical values of CSF can help to predict the development of cerebral aneurysms in late stages and the risk of rupture.法医解剖事例で観察される未破裂脳動脈瘤のうち、従来の小動脈瘤（11 mm未満）と微小動脈瘤（隣接する正常径より約2倍）に分けて、それらの頻度と分布を、脳脊髄液(CSF)の生化学的性状と併せて解析した結果、CSF中のゼラチナーゼ(galatinase A及びgalatinase B)活性が脳動脈瘤早期病変の鋭敏な指標となりうる（p<0.001）事を以前示したが、瘤形成の進展過程におけるCSF中のゼラチン分解活性は、局所のCa調節を司るα-Klotho蛋白の部位別測定値と、少なくとも初期段階では相関していなかった。小動脈~大型動脈瘤さらには巨大動脈瘤の進展及び破裂機序に関わる可能性が示唆された。研究課題/領域番号:18K10120, 研究期間(年度):2018-04-01 - 2022-03-31出典：研究課題「血管壁の極性とプロテアーゼ活性に着目した頭蓋内組織脆弱性の解析」課題番号18K10120（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-18K10120/18K10120seika/）を加工して作

The pronounced lung lesions developing in LATY136F knock-in mice mimic human IgG4-related lung disease

International audienceRationale Immunoglobulin (Ig) G4-related disease (IgG4-RD) is a novel clinical disease entity characterized by an elevated serum IgG4 concentration and tumefaction or tissue infiltration by IgG4-positive plasma cells. Pathological changes are most frequently seen in the pancreas, lacrimal glands, and salivary glands, but pathological changes in the lung also exist. Linker for activation of T cell ( LAT ) Y136F knock-in mice show Th2-dominant immunoreactions with elevated serum IgG1 levels, corresponding to human IgG4. We have reported that LAT Y136F knock-in mice display several characteristic features of IgG4-RD and concluded that they constitute an appropriate model of human IgG4-RD in salivary glands, pancreas, and kidney lesions. Objectives The aim of this study is to evaluate whether lung lesions in LAT Y136F knock-in mice can be a model of IgG4-related lung disease. Methods Lung tissue samples from LAT Y136F knock-in mice (LAT) and wild-type mice (WT) were immunostained for IgG1 and obtained for pathological evaluation, and cell fractions and cytokine levels in broncho-alveolar lavage fluid (BALF) were analyzed. Results In the LAT group, IgG1-positive inflammatory cells increased starting at 4 weeks of age and peaked at 10 weeks of age. The total cell count and percentage of lymphocytes increased significantly in BALF in the LAT group compared to the WT group. In BALF, Th2-dominant cytokines and transforming growth factor-β were also increased. In the LAT group, marked inflammation around broncho-vascular bundles peaked at 10 weeks of age. After 10 weeks, fibrosis around broncho-vascular bundles and bronchiectasis were observed in LAT Y136F knock-in mice but not WT mice. Conclusions LAT Y136F knock-in mice constitute an appropriate model of lung lesions in IgG4-RD

Lat(Y136F) knock-in mouse model for human IgG4-related disease

International audienceBackground The adaptor protein Linker for activation of T cell (LAT) is a key signaling hub used by the T cell antigen receptor. Mutant mice expressing loss-of-function mutations affecting LAT and including a mutation in which tyrosine 136 is replaced by a phenylalanine (Lat(Y136F)) develop lymphoproliferative disorder involving T helper type 2 effector cells capable of triggering a massive polyclonal B cell activation that leads to hypergammaglobulinemia G1 and E and to non-resolving inflammation and autoimmunity. The purpose of this study was to evaluate whether the phenotypes of Lat(Y136F) knock-in mice resemble the immunohistopathological features of immunoglobulin G4-related disease (IgG4-RD).Methods Lat(Y136F) knock-in mice were sacrificed at 4-20 weeks of age, and pancreas, kidney, salivary gland and lung were obtained. All organs were stained with hematoxylin-eosin and with Azan for estimation of collagen in fibrosis, and the severity scores of inflammation and fibrosis were evaluated. Immunostainings were performed to analyze the types of infiltrating cells. In addition, the effects of corticosteroid treatment on the development of tissue lesions and serum levels of IgG1 were assessed.Results Tissue lesions characterized by inflammatory mononuclear cell infiltration and fibrosis were detected in pancreas, kidney, and salivary gland starting from 6 weeks of age. Immunostainings showed pronounced infiltration of plasma cells, CD4-positive T cells, and macrophages. Infiltrating plasma cells predominantly expressed IgG1. The extent of inflammation in pancreas and salivary glands was markedly reduced by corticosteroid treatment.Conclusions Lat(Y136F) knock-in mice displayed increased production of Th2-type IgG1 (a homologue of human IgG4) and developed multiple organ tissue lesions reminiscent of those seen in patients with IgG4-RD. Moreover, the development of these tissue lesions was highly sensitive to corticosteroid treatment like in IgG4-RD. For these reasons we consider the Lat(Y136F) knock-in mouse strain to represent a promising model for human IgG4-RD