- Author:
Jing-Wen DU
1
;
Yu-Xin WANG
2
;
Wei-Jun ZHOU
1
;
Chun-Jun JIANG
1
;
Xiao-Ling XIE
1
;
Hong-Hao ZHANG
1
;
Yan-Jie HE
1
;
Yu-Hua LI
3
Author Information
- Publication Type:Journal Article
- MeSH: Adaptor Proteins, Signal Transducing; immunology; Animals; Epitopes, T-Lymphocyte; metabolism; HLA-A2 Antigen; metabolism; Humans; Mice; Mice, Transgenic; T-Lymphocytes, Cytotoxic
- From: Journal of Experimental Hematology 2016;24(3):865-872
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSOnline 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.
RESULTSFour 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.
CONCLUSIONNatural epitopes (p101-109, p276-284 and p360-368) may be the HLA-A*0201 restricted epitope derived from Eps8.