Zhaojin Yu1,2, Wensi Liu1,2, Ying He1,2,3, Mingli Sun1,2, Jiankun Yu1,2, Xue Jiao1,2, Qiang Han1,4, Haichao Tang1,2, Bing Zhang1,2, Yunkai Xian1,2, Jing Qi1,2, Jing Gong1,2, Wang Xin5, Gang Shi6, Fengping Shan7, Rui Zhang8, Jianping Li9,10,11, Minjie Wei12,13. 1. Department of Pharmacology, School of Pharmacy, China Medical University, No. 13, Beihai Road, Dadong District, Shenyang, Liaoning Province, China. 2. Liaoning Key Laboratory of Molecular Targeted Antitumour Drug Development and Evaluation, Liaoning Cancer Immune Peptide Drug Engineering Technology Research Centre, Key Laboratory of Precision Diagnosis and Treatment of Gastrointestinal Tumours, Ministry of Education, China Medical University, No.77, Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, China. 3. The Third Department of Medical Oncology, The Fourth Hospital of China Medical University, Shenyang City, Liaoning Province, China. 4. Department of Pharmacy, Harrison International Peace Hospital, Hengshui, Hebei Province, China. 5. Liaoning Medical Diagnosis and Treatment R&D Centre Co. Ltd., Shenyang, Liaoning Province, China. 6. Department of Colorectal Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Intitute, No.77, Xiaoheyan Road, Dadong District, Shenyang, Liaoning Province, China. 7. Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, Liaoning Province, China. 8. Department of Colorectal Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Intitute, No.77, Xiaoheyan Road, Dadong District, Shenyang, Liaoning Province, China. zhangrui@cancerhosp-ln-cmu.com. 9. Department of Pharmacology, School of Pharmacy, China Medical University, No. 13, Beihai Road, Dadong District, Shenyang, Liaoning Province, China. ljp_63@163.com. 10. Transfusion Medicine Institute, Liaoning Blood Centre, Shenyang, Liaoning Province, China. ljp_63@163.com. 11. Transfusion Medicine Institute, Harbin Blood Centre, Harbin, Heilongjiang Province, China. ljp_63@163.com. 12. Department of Pharmacology, School of Pharmacy, China Medical University, No. 13, Beihai Road, Dadong District, Shenyang, Liaoning Province, China. mjwei@cmu.edu.cn. 13. Liaoning Key Laboratory of Molecular Targeted Antitumour Drug Development and Evaluation, Liaoning Cancer Immune Peptide Drug Engineering Technology Research Centre, Key Laboratory of Precision Diagnosis and Treatment of Gastrointestinal Tumours, Ministry of Education, China Medical University, No.77, Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, China. mjwei@cmu.edu.cn.
Abstract
BACKGROUND: CD8+ T cell-mediated adaptive cellular immunity and natural killer (NK) cell-mediated innate immunity both play important roles in tumour immunity. This study aimed to develop therapeutic tumour vaccines based on double-activation of CD8+ T and NK cells. METHODS: The immune Epitope database, Molecular Operating Environment software, and enzyme-linked immunosorbent assay were used for epitope identification. Flow cytometry, confocal microscopy, UPLC-QTOF-MS, and RNA-seq were utilized for evaluating immunity of PBMC-derived DCs, CD8+ T or NK cells and related pathways. HLA-A2.1 transgenic mice combined with immunologically reconstituted tumour-bearing mice were used to examine the antitumour effect and safety of epitope vaccines. RESULTS: We identified novel HLA-A2.1-restricted extracellular matrix protein 1(ECM1)-derived immunodominant epitopes in which LA induced a potent immune response. We also found that LA-loaded DCs upregulated the frequency of CD3+/CD8+ T cells, CD45RO+/CD69+ activated memory T cells, and CD3-/CD16+/CD56+ NK cells. We demonstrated cytotoxic granule release of LA/DC-CTLs or LA/DC-NK cells and cytotoxicity against tumour cells and microtissue blocks via the predominant IFN-γ/perforin/granzyme B cell death pathway. Further investigating the mechanism of LA-mediated CD8+ T activation, we found that LA could be internalized into DCs through phagocytosis and then formed a LA-MHC-I complex presented onto the DC surface for recognition of the T cell receptor to upregulate Zap70 phosphorylation levels to further activate CD8+ T cells by DC-CTL interactions. In addition, LA-mediated DC-NK crosstalk through stimulation of the TLR4-p38 MAPK pathway increased MICA/B expression on DCs to interact with NKG2D for NK activation. Promisingly, LA could activate CD8+ T cells and NK cells simultaneously via interacting with DCs to suppress tumours in vivo. Moreover, the safety of LA was confirmed. CONCLUSIONS: LA-induced immune antitumour activity through DC cross-activation with CD8+ T and NK cells, which demonstrated proof-of-concept evidence for the capability and safety of a novel therapeutic tumour vaccine.
BACKGROUND:CD8+ T cell-mediated adaptive cellular immunity and natural killer (NK) cell-mediated innate immunity both play important roles in tumour immunity. This study aimed to develop therapeutic tumour vaccines based on double-activation of CD8+ T and NK cells. METHODS: The immune Epitope database, Molecular Operating Environment software, and enzyme-linked immunosorbent assay were used for epitope identification. Flow cytometry, confocal microscopy, UPLC-QTOF-MS, and RNA-seq were utilized for evaluating immunity of PBMC-derived DCs, CD8+ T or NK cells and related pathways. HLA-A2.1 transgenic mice combined with immunologically reconstituted tumour-bearing mice were used to examine the antitumour effect and safety of epitope vaccines. RESULTS: We identified novel HLA-A2.1-restricted extracellular matrix protein 1(ECM1)-derived immunodominant epitopes in which LA induced a potent immune response. We also found that LA-loaded DCs upregulated the frequency of CD3+/CD8+ T cells, CD45RO+/CD69+ activated memory T cells, and CD3-/CD16+/CD56+ NK cells. We demonstrated cytotoxic granule release of LA/DC-CTLs or LA/DC-NK cells and cytotoxicity against tumour cells and microtissue blocks via the predominant IFN-γ/perforin/granzyme B cell death pathway. Further investigating the mechanism of LA-mediated CD8+ T activation, we found that LA could be internalized into DCs through phagocytosis and then formed a LA-MHC-I complex presented onto the DC surface for recognition of the T cell receptor to upregulate Zap70 phosphorylation levels to further activate CD8+ T cells by DC-CTL interactions. In addition, LA-mediated DC-NK crosstalk through stimulation of the TLR4-p38 MAPK pathway increased MICA/B expression on DCs to interact with NKG2D for NK activation. Promisingly, LA could activate CD8+ T cells and NK cells simultaneously via interacting with DCs to suppress tumours in vivo. Moreover, the safety of LA was confirmed. CONCLUSIONS: LA-induced immune antitumour activity through DC cross-activation with CD8+ T and NK cells, which demonstrated proof-of-concept evidence for the capability and safety of a novel therapeutic tumour vaccine.
Entities:
Keywords:
CTL epitope; DC cross-presentation; DC-NK crosstalk; Extracellular matrix protein 1; Therapeutic tumour vaccine
Authors: Robin Parihar; Charlotte Rivas; Mai Huynh; Bilal Omer; Natalia Lapteva; Leonid S Metelitsa; Stephen M Gottschalk; Cliona M Rooney Journal: Cancer Immunol Res Date: 2019-01-16 Impact factor: 11.151
Authors: Randi Vita; James A Overton; Jason A Greenbaum; Julia Ponomarenko; Jason D Clark; Jason R Cantrell; Daniel K Wheeler; Joseph L Gabbard; Deborah Hix; Alessandro Sette; Bjoern Peters Journal: Nucleic Acids Res Date: 2014-10-09 Impact factor: 16.971