title: [Prediction and Identification of HLA-A*0201 Restricted CTL Epitopes from Eps8].
abstract: OBJECTIVE
  To find and identify HLA-A*0201 restricted cytotoxic T lymphocyte (CTL) epitopes from epidermal growth factor pathway substrate number 8 (Eps8) for specific immunotherapy based on Eps8-derived epitopes in clinic.
  
  
  METHODS
  Online biological softwares involved C-proteasomal cleavage, MHC class I binding affinity and TAP transport efficiency were used for prediction of HLA-A*0201 restricted epitopes from Eps8. Then, T2-binding assays and peptide/MHC complex stability tests were used to further verify the predicted epitopes. Specific secretion of IFN-γ from human CTL was assayed using the IFN-γ ELISPOT kit, and cytolytic activity was measured by a 4-h lactate dehydrogenase (LDH) release assay. Finally, the functional effects in vivo were measured in HLA-A*0201/Kb transgenic (Tg) mice.
  
  
  RESULTS
  Four natural epitopes were designed through online biological softwares. Of the four epitopes selected, p360-368 was found to have the high binding affinity to HLA-A*0201, while p101-109 and p276-284 showed moderate affinities. DC50 of peptide/MHC complexes of the natural epitopes mentioned were all longer than 8 h. In functional assays with human PBMNC in vitro and in HLA-A*0201/Kb transgenic mice in vivo, CTLs primed by each epitope (p101-109, p276-284 and p360-368) secreted IFN-γ and were toxic to cancer cells from a variety of tissue types in an HLA-A*0201-restricted and Eps8-specific manner.
  
  
  CONCLUSIONS
  Natural epitopes (p101-109, p276-284 and p360-368) may be the HLA-A*0201 restricted epitope derived from Eps8.
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China.
authors.forename: Jing-Wen
authors.initials: JW
authors.lastname: Du
authors.affiliation: Second Clinical Medical College of Southern Medical University, Guangzhou 510282, Guangdong Province, China.
authors.forename: Yu-Xin
authors.initials: YX
authors.lastname: Wang
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China.
authors.forename: Wei-Jun
authors.initials: WJ
authors.lastname: Zhou
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China.
authors.forename: Chun-Jun
authors.initials: CJ
authors.lastname: Jiang
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Xie
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China.
authors.forename: Hong-Hao
authors.initials: HH
authors.lastname: Zhang
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China.
authors.forename: Yan-Jie
authors.initials: YJ
authors.lastname: He
authors.affiliation: Department of Hematology, Southern Medical University Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China. E-mail: liyuhua2011gz@163.com.
authors.forename: Yu-Hua
authors.initials: YH
authors.lastname: Li
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000276:immunology*; D000818:Animals; D018984:Epitopes, T-Lymphocyte / Q000378:metabolism*; D015789:HLA-A2 Antigen / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D013602:T-Lymphocytes, Cytotoxic
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C489989:EPS8 protein, human; D018984:Epitopes, T-Lymphocyte; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen