Analysis of host responses to intracellular bacteria and development of cell-mediated gene therapy

宿主对细胞内细菌的反应分析和细胞介导的基因治疗的发展

• 批准号: 15390325
• 负责人: HARA Toshiro
• 金额: $ 9.41万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390325/
• 关键词: intracellular bacteria; macrophage; Mycobacterium; osteopontin; CXCL7; Sendai virus vector; dendritic cell; T-bet; 結核; 細胞免疫療法; IFN-γ; 細胞遺伝子療法; 結核感染マウスモデル

1.Analysis of host responses to intracellular bacteriaGranulocyte-macrophage colony-stimulating factor (GM-CSF)-induced human monocyte-derived macrophage (GM-Mφ) or macrophage CSF (M-CSF)-induced human monocyte-derived Mφ (M-Mφ) are distinct in terms of the resistance to Mycobacterium tuberculosis. To elucidate the role of molecules involved in the functional differences between these Mφs, we investigated the gene expression profiles using microarray. After culture of CD14^+ monocytes with CSFs, Mφs were cultured with or without bacillus Calmette-Guerin (BCG) (GM-Mφ-BCG and M-Mφ-BCG). The gene expression profiles from these cells were compared. Chemokines highly expressed in M-Mφs were selected and evaluated for anti-mycobacterial activity and superoxide production. FN1 and FCGR2B were the most up-regulated genes in GM-Mφ and M-Mφ, respectively. After stimulation with BCG, three chemokine genes (Osteopontin (SPP1), CXC chemokine ligand 7(CXCL7) and CC chemokine ligand 11(CCL11)) were highly expressed in M-Mφ-BCG when compared to those in GM-Mφ-BCG. A significantly increased resistance to M. tuberculosis H37Ra was observed after the stimulation of GM-Mφ with SPP1 or CXCL7. Superoxide production levels of SPP1- or CXCL7-stimulated GM-Mφs were higher than those of GM-Mφs without stimulation. These results indicate that both SPP1 and CXCL7 might have a role in the resistance against mycobacteria, at least in part, through augmenting reactive oxygen intermediate production in Mφs.2.Development of cell-mediated gene therapyTo conventional therapy-resistant TB patients, dendritic cell-mediated gene therapy was planned with Sendai virus vector having T-bet gene, which strongly induces Th1 development of T cells. At present, sufficient IFN-gamma production was not obtained. Further study will be necessary to perform cell-mediated gene therapy.

1.粒细胞-巨噬细胞集落刺激因子诱导的人单核细胞来源巨噬细胞(GM-M-φ)和巨噬细胞来源的人单核细胞来源的M-φ(M-M-φ)对结核分枝杆菌的耐药性不同。为了阐明这些M-φS之间的功能差异所涉及的分子所起的作用，我们利用基因芯片研究了这些基因的表达谱。将巨噬细胞集落刺激因子与CD14~+单核细胞共培养后，将M-φS加入或不加入卡介苗(GM-Mφ-卡介苗和M-Mφ-卡介苗)培养。比较这些细胞的基因表达谱。筛选在M-MφS中高表达的趋化因子，并对其抗分枝杆菌活性和超氧化物歧化产物进行评价。FN1和FCGR2B分别是GM-Mφ和M-Mφ中上调最多的基因。经卡介苗刺激后，M-Mφ-BCG较GM-Mφ-BCG高表达骨桥蛋白(Sp1)、CxC趋化因子配体7(CxCL7)和CC趋化因子配体11(CCL11)基因。用SPP1或CXCL7刺激GM-Mφ后，对结核分枝杆菌H37Ra的耐药性显著增加。SPP_1或CXCL_7刺激的GM-MφS的超氧阴离子产生水平高于未刺激的GM-MφS。这些结果表明，SPP1和CXCL7可能至少部分地通过增加M-φ产生的活性氧中间产物而在抗分枝杆菌中起作用。2.细胞介导的基因治疗的发展对于传统治疗耐药的结核病患者，树突状细胞介导的基因治疗是用带有T-bet基因的仙台病毒载体进行的，它能强烈地诱导T细胞的Th1发育。目前，还没有获得足够的干扰素-γ产量。进行细胞介导的基因治疗还需要进一步的研究。

项目成果

小児科学・新生児学テキスト(免疫疾患)改訂第5版

儿科/新生儿科教科书（免疫疾病）修订版第五版

• 发表时间: 2006
• 作者: Ni R;他7名;Hara T;原 寿郎
• 通讯作者: 原 寿郎

Novel roles of osteopontin and CXC chemokine ligand 7 in the defense against mycobacterial infection

骨桥蛋白和CXC趋化因子配体7在防御分枝杆菌感染中的新作用

• 发表时间: 2007
• 期刊: Clin Exp Immunol 143
• 作者: Khajoee V;Saito M;Takada H;Nomura A;Kusuhara K;Yoshida S;Yoshikai Y;Hara T
• 通讯作者: Hara T

Association study of polymorphisms in SOCS family genes with type diabetes mellitus.

SOCS家族基因多态性与型糖尿病的关联研究

• 期刊: Int J Immunogenet (in press)
• 作者: Ni R;他7名;Hara T
• 通讯作者: Hara T

原 寿郎(分担執筆): "総合アレルギー学:高IgE症候群"南山堂. (2003)

Toshiro Hara（合著者）：“综合过敏学：高 IgE 综合征”Nanzando (2003)。

Ishizaki Y, Nomura A, Kusuhara K, 他6名, Hara T: "Quantification of circulating varicella zoster virus-DNA for the early diagnosis of visceral varicella"J Infect. 47.2. 133-138 (2003)

Ishizaki Y、Nomura A、Kusuhara K 和其他 6 人，Hara T：“用于内脏水痘早期诊断的循环水痘带状疱疹病毒 DNA 定量”J Infect 47.2 (2003)。