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Identification and modification of an HLA-A*0201-restricted cytotoxic T lymphocyte epitope from Ran antigen

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Abstract

Ran is considered to be a promising target for tumor-specific immunotherapy because its protein is exclusively expressed in tumor tissues, though its mRNA can be expressed in most normal tissues. In our study, we obtained four candidate wild-type epitopes designated Ran1, Ran2, Ran3, and Ran4, derived from the Ran antigen with the highest predicted affinity with MHC-I, indicated by affinity prediction plots and molecular dynamics simulation. However, in vitro affinity assays of these epitopes showed only a moderate affinity with MHC-I. Thus, we designed altered peptide ligands (APLs) derived from Ran wild-type epitopes with preferred primary and auxiliary HLA-A*0201 molecule anchor residue replacement. Of the eight tested peptides, the 1Y analog had the strongest binding-affinity and lowest-dissociation rate to HLA-A*0201. Additionally, we investigated the CTLs activities induced by Ran wild-type peptides and the APLs in human PBMCs and in HLA-A*0201/Kb transgenic mice. Ran1 1Y was superior to other APLs and wild-type peptides in eliciting epitope-specific CTL immune responses both in vitro and in vivo. In summary, a wild-type epitope of the tumor-specific antigen Ran, expressed broadly in many tumors, was identified and designated Ran1. An APL of Ran1, Ran1 1Y, was further designed and verified in vitro and in vivo and found to elicit a stronger Ran-specific CTL response, indicating a potential anti-tumor application in the future.

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Acknowledgments

This work was supported by the Major State Basic Research Development Program of China (973 Program) (No. 2007CB512401 and 2007CB512805), the National High Technology Research and Development Program of China (863 Program) (No. 2007AA021603 and 2006AA02A207), the Major Program of National Natural Science Foundation of China (No. 30490241), the General program of National Natural Science Foundation of China (No. 30771950, 30400437, 30872352, and 30571835) and CSTC (2008BB5033).

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Authors and Affiliations

  1. Department of Immunology, Institute of Immunology, Third Military Medical University, 400038, Chongqing, China

    Fan Li, Di Yang, Yiqin Wang, Yijing Deng, Li Wang, Xiaoyun Shang, Bing Ni & Yuzhang Wu

  2. Department of General Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, 400042, Chongqing, China

    Fan Li, Baohua Liu & Weidong Tong

  3. Department of Nephrology, Xinqiao Hospital, Third Military Medical University, 400037, Chongqing, China

    Yiqin Wang

  4. Department of Geriatric Gastroenterology, Chinese PLA General Hospital, 28 FuXing Street, 100853, Beijing, China

    Yijing Deng

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  1. Fan Li
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  2. Di Yang
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Correspondence to Bing Ni or Yuzhang Wu.

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Fan Li, Di Yang, Yiqin Wang, and Baohua Liu contributed equally to this work.

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Li, F., Yang, D., Wang, Y. et al. Identification and modification of an HLA-A*0201-restricted cytotoxic T lymphocyte epitope from Ran antigen. Cancer Immunol Immunother 58, 2039–2049 (2009). https://doi.org/10.1007/s00262-009-0712-9

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  • DOI: https://doi.org/10.1007/s00262-009-0712-9

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