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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2010, Vol. 30 Issue (02): 23-26    
研究快报     
靶向融合蛋白XE-TNFαm2激活并清除潜伏于受感染细胞内的HIV的研
路金芝1,陈伟京1*,丁枫3*,佟薇2,李涛1,蒋虹2,杨晶3,丛喆2,魏强2,黄毅3,王憬惺3,卢圣栋1**
1. 中国医学科学院基础医学研究所
2. 中国医学科学院输血研究所
3. 中国医学科学院实验动物研究所
4. 中国医学科学院中国协和医科大学基础医学研究所
A targeting Fusion Protein XE-TNFαm2 Stimulates Latent HIV to Re-propagate and then Kills the Host Cells with un-mature HIV
1.Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences/Peking Union Medical College,Beijing 100005,China
2.Institute of Laboratory Animal Sciences,CAMS/PUMC, Beijing 100021,China
3.Institute of Blood Transfusion, CAMS, Chengdu 610081,China
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摘要:

制备了工程化靶向融合蛋白XE-TNFαm2。其中,XE为HIV/SIV辅助受体CXCR4的第二胞外域。TNFαm2是经突变改型的TNFα,其毒副作用已降低18倍,己用于临床治疗恶性肿瘤。本研究所用的整合有HIV的标准细胞株J-Lat Tat-GFP(H2/9855),为美国NIH艾滋病试剂中心所赠送。其中,一个经缺失突变后的HIV被整合在Jurkat细胞的染色体上,成为5’LTR-Tat-GFP-3’LTR。不同剂量的XE-TNFαm2加于一定量的JurkatH2/9855细胞后,流式细胞仪检测结果表明,荧光蛋白的表达量随着处理时间的延续而增加,并有XE-TNFαm2剂量的依赖关系。这一结果表明,XE-TNFαm2可强力激活潜伏于细胞染色体中的HIV,使之重新繁殖起来。鉴于己有的研究表明,XE-TNFαm2可杀灭受HIV/SIV感染的细胞。据此,当重新繁殖的HIV开始出芽时,其gp120必然出现在宿主细胞表面,且此gp120必然被XE-TNFαm2中的XE所结合,并其TNFαm2的杀伤信号将转导进入细胞。这样,这些宿主细胞将被杀灭。细胞的死亡导致未成熟HIV繁殖的中止。最后,在重新繁殖且成熟起来的HIV导致细胞破碎并释放出细胞之前,细胞内尚无感染力的未成熟HIV将同死亡的宿主细胞一起被清除。

关键词: 靶向融合蛋白激活HIV    
Abstract:

Engineered targeting fusion protein XE-TNFαm2 was prepared.In which,XE is the second excellular domain of CXCR4,a co-receptor of HIV/SIV. TNFαm2 is a mutation protein of TNFα.Its side effect was reduced 18 fold.It has been used in clinic for cancer therapy.In this study, Jurkat H2/9855, a standard cell line was donated toward by AIDS Reagent Program, NIH ,USA.In which, a deleted HIV mutant was inserted into chromosome of Jurkat cells to be 5’LTR-Tat-GFP-3’LTR. XE-TNFαm2 with different doses were added into 1x105/ml Jurkat H2/9855 cell suspensions respectively. After 24hrs,48 hrs and 72 hrs,the GFP in samples were determined by flow cytometry. The results showed that the increase of GFP expression level is along with the extended time of treatment with XE-TNFαm2,and it is depended on the dosage of XE-TNFαm2. This result indicated that XE-TNFαm2 is able to stimulate latent HIV to re-propagate in Jurkat H2/9855 cells.Thus,the host cell were became to be novel infected cells. On the other aspect, our previous study indicated that one of XE-TNFαm2 functions is to kill the cells infected by HIV/SIV. Accordingly, when the novel re-propagating HIVs start to pullulate, their gp120 must be appeared on the surface of host cells. Then,gp120 must be bound by XE in XE-TNFαm2, and latter’s killing signal must be transduced into cells.In that case, these host cells must be killed by XE-TNFαm2. Thus, the death of host cells leads to stop the propagation of un-maturated HIVs. Then, such kind of HIV without infection ability must be eliminated together with died host cells before that the re-propagated HIVs maturated to break host cells and to release out from cells.

Key words: Targeting    Fusion protein    Stimulate HIV
收稿日期: 2010-02-01 出版日期: 2010-02-26
通讯作者: 卢圣栋     E-mail: lusd@pumc.edu.cn
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引用本文:

路金芝 陈伟京 丁枫 佟薇 李涛 蒋虹 杨晶 丛喆 魏强 黄毅 王憬惺 卢圣栋. 靶向融合蛋白XE-TNFαm2激活并清除潜伏于受感染细胞内的HIV的研[J]. 中国生物工程杂志, 2010, 30(02): 23-26.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I02/23

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