Modeling of the inherence of feedback regulation and stem cell behavior in granulopoiesis

金沢大学医学部附属病院小児科Long-standing controversies in hematopoiesis are the mechanisms of self-maintenance and differentiation commitment of the hematopoietic stem cells (HSC), and regulation of the peripheral control of hematopoiesis. In the present study, we have applied a threedimensional cellular automaton (CA) model to granulopoiesis in order to identify the internally generative theoretical relationship between microscopic mechanisms and macroscopic behavior of hematopoietic processes. The number of mitotic event of the cells in a proliferating phase, the transit time of each of 15 differential stages from HSC to mature cells (designated as Tl to Tl 5, and Tdup for HSC duplication time), and the neighborhood rules for HSC self-renewal were incorporated in this model system as analytical parameters. Homeostatic granulopoiesis was achieved when the following inequalities for the transit times were fulfilled: Tl > Z Tn (n = 2 to 15) and Tdup > 1/2 Tl. Importantly, stabilization of the cell production was induced in a negative feedback manner following external perturbation of the peripheral granulocyte numbers. The Tdup of individual HSC was dramatically fluctuated to produce the offspring responding to this perturbation. A single cell kinetic analysis demonstrated that symmetrical or asymmetrical cell division of the HSC was recruited in a transitional manner resulting in generation of the regulatory effect on the lineage-commitment processes. The inherence of feedback regulation would be a characteristic feature of the emergent dynamical property in the hematopoietic system. The CA modeling will provide the framework to analyze the behavior of HSC and to understand abnormal kinetics of the cells such as minimal residual disease in the treatment of leukemias

Intermittent X-linked thrombocytopenia with a novel WAS gene mutation

X-linked thrombocytopenia (XLT) is caused by mutations in the WAS gene and characterized by thrombocytopenia with minimal or no immunodeficiency. Patients with XLT usually exhibit persistent thrombocytopenia, and intermittent thrombocytopenia has been described only in two families. Here, we report a patient with intermittent XLT carrying a novel missense mutation (Ala56Thr). He showed residual expression of Wiskott-Aldrich syndrome protein in the lymphocytes and platelets. There appeared to be an association between normal platelet numbers and a post infectious state. Our findings further support the importance of analysis of Wiskott-Aldrich syndrome protein in male patients who exhibit fluctuating courses of thrombocytopenia. Pediatr Blood Cancer 2014;61:746-748. © 2013 Wiley Periodicals, Inc

Phase II study of S-1 on alternate days plus bevacizumab in patients aged ≥ 75 years with metastatic colorectal cancer (J-SAVER)

BackgroundAlternate-day administration of S-1 is thought to reduce toxicities. This phase II study evaluated S-1 on alternate days combined with bevacizumab as first-line treatment for elderly patients with metastatic colorectal cancer.Patients and methodsEligible patients had histologically proven colorectal adenocarcinoma, measurable metastatic lesions, age ≥ 75 years, Eastern Cooperative Oncology Group performance status ≤ 1, no previous chemotherapy, and refused oxaliplatin- or irinotecan-containing regimens. Patients received 40 mg, 50 mg, or 60 mg (body surface area ≤ 1.25 m2, > 1.25 to ≤ 1.50 m2, or > 1.50 m2, respectively) of S-1 twice orally on Sunday, Monday, Wednesday, and Friday every week. Bevacizumab (7.5 mg/kg) was administered every 3 weeks. The primary endpoint was progression-free survival.ResultsOf 54 enrolled patients, 50 patients were evaluated for efficacy and 53 for safety. The median age was 79 years (range 75–88 years). The median progression-free survival was 8.1 months (95% confidence interval (CI) 6.7–9.5 months). The median overall survival was 23.1 months (95% CI 17.4–28.8 months). The response rate was 44% (95% CI 30.2–57.8%), and the disease control rate was 88% (95% CI 79.0–97.0%). Grade 3 or higher hematologic, non-hematologic, and bevacizumab-related adverse events occurred in 9%, 11%, and 25% of patients, respectively. The most common grade 3 and 4 treatment-related adverse events were hypertension (11%), nausea (6%), fatigue (6%), anemia (6%), and proteinuria (6%). Only 6 patients discontinued treatment due to adverse events.ConclusionS-1 on alternate days combined with bevacizumab showed better tolerability and comparable survival compared with the results of similar studies

Reactive peripheral blood plasmacytosis in a patient with acute hepatitis A

金沢大学附属病院小児科Reactive plasmacytosis is a transient expansion of plasma cell progenitors and precursors. This rare condition has been reported to occur mainly in infections and tumors. We describe a case of acute hepatitis A presenting with marked peripheral blood plasmacytosis. Plasma cells made up 27.5% of the mononuclear cells and had the immunophenotype CD10-CD19 +CD20-CD21-CD23-CD34 -CD38++HLA-DR+. Although the level of interleukin 6 was not increased, the presence of activated T-cells with an inverted CD4/CD8 ratio and high levels of soluble interleukin 2 receptor and neopterin indicated a marked immune response to acute hepatitis A. The patient\u27s plasma cells had almost disappeared from the blood by hospital day 16. This report may represent the first described case of reactive peripheral blood plasmacytosis in acute hepatitis A. © 2007 The Japanese Society of Hematology

制御性樹状細胞の大量培養を用いた新たな造血幹細胞移植の開発

マウス骨髄細胞からGM-CSFを添加し未熟樹状細胞を大量に培養した後、psolarenと紫外線を併用し(いわゆるPUVA療法の応用)制御性樹状細胞を大量に作製することに成功した。その細胞は、混合リンパ球反応(MLR)においてMHC非拘束性に抑制性の機能を有していた。メカニズムは、抗炎症性サイトカイン(IL-10やTGF-β)によるものではなく、細胞間の接触によって引き起こされていた。トリプトファン代謝による細胞増殖の抑制をみるため、indoleamine 2,3-dioxygenase (IDO)の発現を定量PCR法にて測定したところ、骨髄由来の樹状細胞に比べて5倍以上発現していた。IDOを発現し、MHC非拘束性に抑制機能を有するPUVA処理した樹状細胞による細胞療法は、致死的なGVHDに対して有用な治療法と成りうる可能性がある。Bone marrow-derived cultured DCs acquired tolerogenicity by PUVA (psoralen+UVA) treatment in mice. In MLR assays, strong tolerogenicity was observed when adding PUVA-DCs generated from the same strain of stimulator cells, or responder cells, or even from third party strain into MLR mixture. That is, the PUVA-treated DCs have tolerogenic function in a MHC-independent manner, in part, due to the reduced expression levels of CD80 and CD86. To clarify the mechanisms of how PUVA-treated DCs induce tolerogenicity in further detail, we performed MLR with the addition of neutralizing antibodies against IL-10 or TGF-b1 or both. Neutralization of immunosuppressive cytokines had no effects on MLR. We then showed that cell-to-cell contact between PUVA-DCs and alloreactive T-cells was needed to mediate the regulatory effect by transwell experiment. Next we compared the expression of the indoleamine 2,3-dioxygenase (IDO), which induces T-cell anergy by tryptophan depletion and by the production of metabolic byproducts collectively known as kynurenines, by real-time PCR between PUVA-DCs and BM-DCs. An increase IDO gene transcription level was observed in PUVA-DCs about 5 times more than in BM-DCs (p<0.01). In conclusion, PUVA-DCs acquired regulatory function in a MHC-independent manner by upregulation of IDO. Infusion of PUVA-treated DCs could have a great potential to treat lethal acute GVHD in clinical settings

PUVA処理した制御性樹状細胞の更なる免疫抑制増強の試み: 新たなGVHD治療戦略

金沢大学附属病院PUVA処理（Psoralen+UVA照射2J）した樹状細胞(PUVA-DC)がin vitroにおいて、制御性樹状細胞の性質を獲得したことを報告してきた．その細胞をマウスGVHDモデルにて検討を行ったが、GVHD抑制作用が得られなかったため、PUVA-DCの作製条件の検討を行った。UVA照射の設定（0-2 J）を探索したが、0.2-1Jでは免疫寛容を有していなかった。次にカスパーゼ阻害剤による処理を追加したが、PUVA-DCの24時間後の生存率は改善せず免疫寛容誘導能も有していなかった．PUVA-DCを、細胞治療に用いるためには、さらなる培養条件の検討が必要である．We have reported that bone marrow-derived dendritic cells (BM-DCs), which are generated from bone marrow cells plus GM-CSF, acquired tolerogenicity by PUVA-treatment(psoralen+UVA 2J/cm2) in mice.The PUVA-treated DCs have tolerogenic function in a MHC-independent manner. But these cells did not have the inhibitory effects in the mouse GVHD model. We tried to improve the manufacture condition of PUVA-DC, to make these cells engraft effectively in vivo. At first, we researched the setting (0-2 J /cm2) of the UVA radiation, but did not have acquired tolerance in the UVA 0.2-1J/cm2 setting. Next, we added caspase inhibitor to PUVA treatment for apoptotic suppression by the UVA radiation. The survival rate 24 hours after PUVA-treatment were not improved, and these cells did not have the acquired more tolerance than conventional PUVA-DC. The examination of the further culture condition is necessary to use PUVA-DC for cell therapy.研究課題/領域番号:17K10135, 研究期間(年度):2017-04-01 – 2020-03-31出典：「PUVA処理した制御性樹状細胞の更なる免疫抑制増強の試み: 新たなGVHD治療戦略」研究成果報告書　課題番号17K10135（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/report/KAKENHI-PROJECT-17K10135/17K10135seika/)を加工して作