Immunohematologic studies on the SDF-1/CXCR-4 system with monoclonal antibod.

使用单克隆抗体对 SDF-1/CXCR-4 系统进行免疫血液学研究。

• 批准号: 09671107
• 负责人: HORI Toshiyuki
• 金额: $ 1.86万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671107/
• 关键词: CXCR-4; CDF-1; chemokine; HIV-1; CXCR4; モノクローナル抗体; HIV

Three mAbs, termed IVR7, AI58, and THS123, were generated and demonstrated to react specifically with human CXCR-4-transfected COS-7 cells. THSl23 could immunoprecipitate a component with an apparent mol wt of 47 kD from CXCR-4^+ cells. Flow cytometric analysis showed that most of the hematopoietic cell lines and some non-hematopoietic cell lines expressed CXCR-4. A fraction of normal peripheral blood mononulcear cells expressed CXCR-4 but neutrophils were negative. Two color analysis revealed that the majority of but not all T cells, virtually all B cells and all moncytes expressed CXCR-4 while it was hardly detectable on NK cells. As to differentiated helper T cells, Th2 but not Th1 expressed CXCR-4. It was also found that IL-4 could induce expression of functional CXCR-4 on Th1. Thus, expression of CXCR-4 is not ubiquitous but rather cell type-specific in hematopoietic cells.Entry of HIV-1 is initiated by interaction of the envelope protein gp120 with CD4 and one of the relevant chemokine receptors. Although the V3 region of gp120 is speculated to be involved in interaction with chemokine receptors, it is still unclear if the V3 region can directly bind to them. Using synthetic V3 peptides that correspond to the V3 regions of gp120 of T-tropic, M-tropic and dual tropic HIV-1, we could demonstrate that V3 peptides of T-tropic and dual tropic strains but not that of an M-tropic strain could directly bind to CXCR-4, which indicate that the V3 region of gp120 can bind to the relevant chemokine receptor by itself without CD4 or other domains of gp120.

产生了三种mAb，称为IVR 7、AI 58和THS 123，并证明其与人CXCR-4转染的COS-7细胞特异性反应。THS 123可免疫沉淀CXCR-4^+细胞中表观分子量为47 kD的组分。流式细胞术分析显示，大多数造血细胞系和一些非造血细胞系表达CXCR-4。一部分正常外周血单核细胞表达CXCR-4，而中性粒细胞为阴性。双色分析显示，大多数但不是所有的T细胞、几乎所有的B细胞和所有的单核细胞表达CXCR-4，而在NK细胞上几乎检测不到。在分化的辅助性T细胞中，Th 2表达CXCR-4，而Th 1不表达。IL-4可诱导Th 1细胞表达功能性CXCR-4。因此，CXCR-4的表达在造血细胞中并非普遍存在，而是具有细胞类型特异性。HIV-1的进入是通过包膜蛋白gp 120与CD 4和相关趋化因子受体之一的相互作用启动的。尽管推测gp 120的V3区参与与趋化因子受体的相互作用，但仍不清楚V3区是否可以直接与它们结合。使用与嗜T、嗜M和双嗜性HIV-1的gp 120的V3区相对应的合成V3肽，我们可以证明嗜T和双嗜性毒株的V3肽而不是嗜M毒株的V3肽可以直接结合CXCR-4，这表明gp 120的V3区可以在没有CD 4或gp 120的其他结构域的情况下自身结合相关趋化因子受体。

项目成果

T.Hori: "Delineation of CXCR-4 as an entry cofactor for T-tropic HIV-1 by monoclonal anfibodies" Immunology Letters. 56. 15-15 (1997)

T.Hori：“单克隆抗体将 CXCR-4 描述为 T 向性 HIV-1 的进入辅助因子”免疫学快报。

Patrick Jourdan: "Interleukin-4 induces functional expression of CXCR-4 on human Th1 and Th2 cells which results in activation of ERK-2 MAP kinase" The Journal of Immunology. 160. 4153-4157 (1998)

Patrick Jourdan：“Interleukin-4 诱导人类 Th1 和 Th2 细胞上 CXCR-4 的功能性表达，从而激活 ERK-2 MAP 激酶”《免疫学杂志》。

Hitoshi Sakaida, Toshiyuki Hori, Akihito Yonezawa, Akihiko Sato, Yoshitaka Isaka, Osamu Yoshie, Toshio Hattori, and Takashi Uchiyama.: "T-tropic human immunodeficiency virus type 1 (HIV-1)-derived V3 loop peptides directly bind to CXCR-4 and inhibit T-tro

Hitoshi Sakaida、Toshiyuki Hori、Akihito Yonezawa、Akihiko Sato、Yoshitaka Isaka、Osamu Yoshie、Toshio Hattori 和 Takashi Uchiyama。：“T-tropic 人类免疫缺陷病毒 1 型 (HIV-1) 衍生的 V3 环肽直接与 CXCR- 结合

Huiling Hu, Tatsuo Shioda, Toshiyuki Hori, Chiakaya Moriya, Atsushi Kato, Yuko Sakai, Kouji Matsushima, Takashi Uchiyama, and Yoshiyuki Nagai.: "Internalization of CXCR-4 is not required for its coreceptor activity for HIV-1 entry." Archives of Virology.

Huiling Hu、Tatsuo Shioda、Toshiyuki Hori、Chiakaya Moriya、Atsushi Kato、Yuko Sakai、Kouji Matsushima、Takashi Uchiyama 和 Yoshiyuki Nagai.：“CXCR-4 的内化并不是其辅助受体活性进入 HIV-1 所必需的。”

M.Arai: "Human T-cell leukemia virus type I Tax protein induces the expression of lymphocyte chemo atlracf ant SDF-1/PBSF" Virofogy. (発表予定).

M.Arai：“人类 T 细胞白血病病毒 I 型 Tax 蛋白诱导淋巴细胞趋化因子 SDF-1/PBSF 的表达”Virofogy（待提交）。