Isolation of apoptosis-related genes induced by CD40 ligation

CD40 连接诱导的凋亡相关基因的分离

• 批准号: 08839018
• 负责人: WANG Jiyang
• 金额: $ 1.41万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08839018/
• 关键词: Anti-IgM antibody; B lymphocyte; Fas expression; Apoptosis; CD40; CD40 ligand; zinc fingerモチーフ

We have found that Fas expression in B cells can be strongly induced by CD40 ligation but not by anti-IgM stimulation of IL-4. We also demonstrated that the susceptibility to Fas-mediated apoptosis of B cells is enhanced by CD40 crosslinking but decreased by treatment with anti-IgM antibodies. Using semi-quantitative RT PCR analysis of total RNA from CD40-activated splenic B cells, we could not find a good correlation between the enhanced Fas susceptibility in B cells stimulated by CD40L and the expression of a couple of apoptosis-related genes including HCP,RIP,FADD and ICE.While CD40 ligation did not affect the expression of bcl-2, bcl-xL and bax, anti-IgM treatment strongly induced bcl-xL expression without altering the expression of bcl-2 or bax. Using a subtractive screening strategy, we have successfully isolated a novel cDNA encoding a 587 a.a.protein. This novel protein contains 15 zinc finger motifs which may form a DNA-binding domain, and an acidic domain at its N-terminal which is a potentially transcriptional activation domain. In addition, we have also succeeded in isolating two other novel genes that are induced by CD40L alone or CD40L plus anti-IgM,respectively. We are currently investigating the potential roles of these novel genes in the B cell activation, Fas induction and Fas-triggered apoptosis, as well as the generation of memory B cells.

我们发现，CD40连接可以强烈诱导B细胞中的FAS表达，而不是通过抗IGM刺激IL-4来诱导。我们还证明，CD40的交联增强了对FAS介导的B细胞凋亡的敏感性，但通过抗IGM抗体的处理降低。 Using semi-quantitative RT PCR analysis of total RNA from CD40-activated splenic B cells, we could not find a good correlation between the enhanced Fas susceptibility in B cells stimulated by CD40L and the expression of a couple of apoptosis-related genes including HCP,RIP,FADD and ICE.While CD40 ligation did not affect the expression of bcl-2, bcl-xL and bax, anti-IgM治疗强烈诱导BCL-XL表达，而不会改变Bcl-2或Bax的表达。使用减法筛选策略，我们成功地隔离了编码587 A.A.蛋白质的新型cDNA。这种新型蛋白质包含15个锌指基序，可能形成一个DNA结合结构域，以及其N末端的酸性结构域，这是一个潜在的转录激活结构域。此外，我们还成功地隔离了单独由CD40L或CD40L加抗IGM诱导的另外两个新型基因。我们目前正在研究这些新基因在B细胞激活，FAS诱导和FAS触发的凋亡以及记忆B细胞的产生中的潜在作用。

项目成果

Koizumi,T.,Wang,J.et al: "Regulation of bcl-XL expression and Fas susceptibility in mouse B cellsby CD40 ligation,surface IgM crosslinking and IL-4." Mol.Immunol.(in press). (1997)

Koizumi,T.,Wang,J.等人：“通过 CD40 连接、表面 IgM 交联和 IL-4 调节小鼠 B 细胞中的 bcl-XL 表达和 Fas 敏感性。”

J.Wang,T.Koizumi,and T.Watanabe: "Altered antigen receptor signaling and impaired Fas-mediated apoptosis of B cells in Lyn-deficient mice." J.Exp.Med.184. 831-838 (1996)

J.Wang、T.Koizumi 和 T.Watanabe：“Lyn 缺陷小鼠中抗原受体信号传导发生改变，Fas 介导的 B 细胞凋亡受损。”

Koizumi, T., Wang, J.et al: "Regulation of bcl-XL expression and Fas susceptibility in mouse B cellsby CD40 ligation, surface IgM crosslinking and IL-4." Mol.Immunol.33. 1247-1253 (1996)

Koizumi, T.、Wang, J.等人：“通过 CD40 连接、表面 IgM 交联和 IL-4 调节小鼠 B 细胞中的 bcl-XL 表达和 Fas 敏感性。”

K.Masuda,J.Wang and T.Watanabe: "Reduced susceptibility to Fas-mediated apoptosis in B1 cells." Immunology. 27. 449-455 (1997)

K.Masuda、J.Wang 和 T.Watanabe：“降低 B1 细胞对 Fas 介导的细胞凋亡的敏感性。”

D-M Su, J.Wang, Q.Lin, et al: "Interferons alpha/beta inhibit IL-7 induced proliferaton of CD4-8-CD3-(TN)CD44+25+thymocytes, but not block that of TN CD44-25-cells." Immunology. 90. 543-549 (1997)

D-M Su、J.Wang、Q.Lin 等人：“干扰素 α/β 抑制 IL-7 诱导的 CD4-8-CD3-(TN)CD44 25 胸腺细胞增殖，但不阻断 TN CD44-25 细胞的增殖