Reconstitution of human lymphoid tissue in mouse and its application

小鼠人淋巴组织的重建及其应用

• 批准号: 12680806
• 负责人: KOYANAGI Yoshio
• 金额: $ 2.37万
• 依托单位: Tohoku University Graduate School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680806/
• 关键词: SCID mouse; lymphoid organ; CD4+ T cell; Apoptosis; HIV; TRAIL; FasL; chemokine; NOD-SCIDマウス; 移植マウス; CD4陽性T細胞破壊; 脾臓; リンパ節構造

We previously reported the establishment of novel mouse strains with defective T and B cells as well as innate immunological functions associated with NK, macrophage, and complement and showed that all mutant mouse strains efficiently received human PBL (peripheral blood leukocyte) engraftment (hu-PBL-scid-mouse) (J. Virol. 71 : 2417-2424, 1997). Of these chimeric mice, hu-PBL-NOD-scid mice, efficient HIV-1 infection was observed after intraperitoneal injection. High levels of viremia (more than 1〜100 ng/ml of gag p24) were obtained by infection with a macrophage-tropic HIV-1 strain, JR-FL. Peak levels of the viremia were observed at 2 weeks post infection and the antigenemia disappeared 4 weeks after infection. In these HIV-1 infected mice, human CD3+/CD4+ and CD3+/CD8+ T cells proliferated up to 2 weeks after infection. In mouse spleen especially, white pulps were reconstructed and HIV_<gag> p24+ cells accumulated in the region where apoptotic cells were also observed. In addition to human T cells, human CD68+ macrophages were also reconstituted in the mouse spleen. These results suggested that the hu-PBL-NOD-scid mouse would be useful to investigate HIV-1 pathogenesis in tissues where human T cells and macrophages coexist. Thus, we started to examine whether the specific signals through molecules such as FasL, TNF, or/and TRAIL are involved in the pathogenesis. Double staining with TUNEL and anti-FasL, anti-TNF, or anti-TRAIL, respectively, revealed that the apoptotic cells were frequently found in conjugation with TRAIL+ cells. On the other hand, FasL+ cells or TNF+ cells were not associated with the apoptotic cells. Further analysis confirmed that the apoptotic and the adjacent TRAIL+ cells were both CD4+ human cells. These results suggest that, in the absence of specific immunity, many of the HIV-1 uninfected CD4+ T cells undergo apoptosis through the TRAIL signal in HIV-1 infected lymphoid organ during primary viremia.

我们之前报道了具有缺陷的T和B细胞以及与NK、巨噬细胞和补体相关的先天免疫功能的新小鼠株的建立，并表明所有突变小鼠株都有效地接受了人外周血白细胞（PBL）移植（hu-PBL-scid-mouse） （J.病毒学报，71:2417-2424,1997）。在这些嵌合小鼠，hu-PBL-NOD-scid小鼠中，腹腔注射后观察到有效的HIV-1感染。感染嗜巨噬细胞的HIV-1毒株JR-FL可导致高水平的病毒血症（gag p24高于1 ~ 100 ng/ml）。病毒血症在感染后2周达到高峰，抗原血症在感染后4周消失。在这些HIV-1感染的小鼠中，人类CD3+/CD4+和CD3+/CD8+ T细胞在感染后2周内增殖。特别是在小鼠脾脏中，重建了白色浆体，在细胞凋亡的区域内聚集了hiv <gag> p24+细胞。除人T细胞外，还在小鼠脾脏中重建了人CD68+巨噬细胞。这些结果表明，hu-PBL-NOD-scid小鼠将有助于研究人类T细胞和巨噬细胞共存的组织中HIV-1的发病机制。因此，我们开始研究通过FasL、TNF或/和TRAIL等分子传递的特定信号是否参与了发病机制。TUNEL和抗fasl、抗tnf、抗TRAIL双染色显示，凋亡细胞多与TRAIL+细胞结合。另一方面，FasL+细胞或TNF+细胞与凋亡细胞无相关性。进一步分析证实，凋亡细胞和相邻的TRAIL+细胞均为CD4+人细胞。这些结果表明，在缺乏特异性免疫的情况下，许多HIV-1未感染的CD4+ T细胞在原发性病毒血症期间通过TRAIL信号在HIV-1感染的淋巴器官中发生凋亡。

项目成果

Miura Y, Misawa N, Maeda N, Inagaki Y, Tanaka Y, Ito M, Kayagaki N, Yamamoto N, Yagita H, Mizusawa H, Koyanagi Y: "Critical contribution of TNF-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4^+ T cells in HIV-1-infected hu-PBL-NOD-SCID

Miura Y、Misawa N、Maeda N、Inagaki Y、Tanaka Y、Ito M、Kayagaki N、Yamamoto N、Yagita H、Mizusawa H、Koyanagi Y：“TNF 相关凋亡诱导配体 (TRAIL) 对细胞凋亡的关键贡献

Ishikawa K, Janssens W, Banor JS, Shinno T, Piedate J, Sata T, Ampofo WK, Brandful J, Koyanagi Y, Yamanoto N, Canas-Ferreira WA, Adu-Sarkodie Y, Kurata T.: "Genetic Analysis of HIV Type2 from Ghana and Guinea-Bissau, West Africa"AIDS Research and Human Re

Ishikawa K, Janssens W, Banor JS, Shinno T, Piedate J, Sata T, Ampofo WK, Brandful J, Koyanagi Y, Yamanoto N, Canas-Ferreira WA, Adu-Sarkodie Y, Kurata T.：“HIV 2 型的基因分析

Tsunetsugu-Yokota Y: "Transcriptional regulation of HIV-1 LTR during antigen-dependent activation of primary T-cell by dendritic cells"Journal of Leukocyte Biology. 67. 432-440 (2000)

Tsunetsugu-Yokota Y：“树突状细胞抗原依赖性激活原代 T 细胞期间 HIV-1 LTR 的转录调节”白细胞生物学杂志。

Miura Y, Misawa N, Maeda N, Inagaki Y, Tanaka Y, Ito M, Kayagaki N, Yamamoto N, Yagits H, Mizusawa H, Koyanagi Y: "Critical contribution of TNF-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4^+T cells in HIV-1-infected hu-PBL-NOD-SCID

Miura Y、Misawa N、Maeda N、Inagaki Y、Tanaka Y、Ito M、Kayagaki N、Yamamoto N、Yagits H、Mizusawa H、Koyanagi Y：“TNF 相关凋亡诱导配体 (TRAIL) 对细胞凋亡的关键贡献

Takahashi Y, Tanaka Y, Yamashita A, Koyanagi Y, Nakamura M, Yamamoto N: "OX40 stimulation by gp34/OX40 ligand enhances productive HIV-1 infection"Journal of Virology. 75. 6748-6757 (2001)

Takahashi Y、Tanaka Y、Yamashita A、Koyanagi Y、Nakamura M、Yamamoto N：“gp34/OX40 配体的 OX40 刺激增强了有效的 HIV-1 感染”病毒学杂志。