一种靶向癌细胞中Fn14受体的基于PE38的重组免疫毒素的生物信息学预测与实验验证

Bioinformatic prediction and experimental validation of a PE38-based recombinant immunotoxin targeting the Fn14 receptor in cancer cells.

作者信息

Keshtvarz Maryam, Salimian Jafar, Yaseri Mehdi, Bathaie Seyedeh Zahra, Rezaie Ehsan, Aliramezani Amir, Norouzbabaei Zahra, Amani Jafar, Douraghi Masoumeh

机构信息

Division of Microbiology, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Chemical Injuries Research Center, Baqiyatallah, University of Medical Sciences, Tehran, Iran.

出版信息

Immunotherapy. 2017 Mar;9(5):387-400. doi: 10.2217/imt-2017-0008.

DOI:10.2217/imt-2017-0008

PMID:28357912

Abstract

AIM

AFn14R can serve as an ideal target for cancer immunotherapy. Here, a combined bioinformatic and experimental approach was applied to characterize an immunotoxin consisting of single-chain variable fragment antibody that targets Fn14 and a toxin fragment (PE38).

METHODS & RESULTS: Flow cytometry results showed that the rate of PE38-P4A8 binding to Fn14 was approximately 60 and 40% in HT-29 and A549 cells, respectively. Moreover, 1 ng/µl of immunotoxin was able to lyse approximately 53 and 41% of HT-29 and A549, respectively. PE38-P4A8 showed stability in mouse serum (∼90%) after 3-h incubation. Most importantly, using bioinformatics for determining the structure and function of fusion proteins can be very helpful in designing of experiments.

CONCLUSION

Coupled with bioinformatics, experimental approaches revealed that PE38-P4A8 could be used as a promising therapeutic agent for cancer cells expressing Fn14.

摘要

目的

AFn14R可作为癌症免疫治疗的理想靶点。在此，采用生物信息学与实验相结合的方法来表征一种免疫毒素，该免疫毒素由靶向Fn14的单链可变片段抗体和毒素片段（PE38）组成。

方法与结果

流式细胞术结果显示，在HT-29和A549细胞中，PE38-P4A8与Fn14的结合率分别约为60%和40%。此外，1 ng/µl的免疫毒素能够分别裂解约53%和41%的HT-29和A549细胞。孵育3小时后，PE38-P4A8在小鼠血清中显示出稳定性（约90%）。最重要的是，利用生物信息学确定融合蛋白的结构和功能对实验设计非常有帮助。