Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma
Background & Aims: Antigen-specific immunotherapy is a promising strategy to treat HBV infection and hepatocellular carcinoma (HCC). To facilitate killing of malignant and/or infected hepatocytes, it is vital to know which T cell targets are presented by human leucocyte antigen (HLA)-I compl...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-11-01
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| Series: | JHEP Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589555922001483 |
| _version_ | 1811235058233638912 |
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| author | Monique T.A. de Beijer Karel Bezstarosti Robbie Luijten Wouter A.S. Doff Patrick P.C. Boor Roel F.A. Pieterman Rachid Bouzid Paula J. Biesta Jan N.M. Ijzermans Michail Doukas Robert A. de Man Andrea M. Woltman Jeroen A.A. Demmers Sonja I. Buschow |
| author_facet | Monique T.A. de Beijer Karel Bezstarosti Robbie Luijten Wouter A.S. Doff Patrick P.C. Boor Roel F.A. Pieterman Rachid Bouzid Paula J. Biesta Jan N.M. Ijzermans Michail Doukas Robert A. de Man Andrea M. Woltman Jeroen A.A. Demmers Sonja I. Buschow |
| author_sort | Monique T.A. de Beijer |
| collection | DOAJ |
| description | Background & Aims: Antigen-specific immunotherapy is a promising strategy to treat HBV infection and hepatocellular carcinoma (HCC). To facilitate killing of malignant and/or infected hepatocytes, it is vital to know which T cell targets are presented by human leucocyte antigen (HLA)-I complexes on patient-derived hepatocytes. Here, we aimed to reveal the hepatocyte-specific HLA-I peptidome with emphasis on peptides derived from HBV proteins and tumour-associated antigens (TAA) to guide development of antigen-specific immunotherapy. Methods: Primary human hepatocytes were isolated with high purity from (HBV-infected) non-tumour and HCC tissues using a newly designed perfusion-free procedure. Hepatocyte-derived HLA-bound peptides were identified by unbiased mass spectrometry (MS), after which source proteins were subjected to Gene Ontology and pathway analysis. HBV antigen and TAA-derived HLA peptides were searched for using targeted MS, and a selection of peptides was tested for immunogenicity. Results: Using unbiased data-dependent acquisition (DDA), we acquired a high-quality HLA-I peptidome of 2 × 105 peptides that contained 8 HBV-derived peptides and 14 peptides from 8 known HCC-associated TAA that were exclusive to tumours. Of these, 3 HBV- and 12 TAA-derived HLA peptides were detected by targeted MS in the sample they were originally identified in by DDA. Moreover, 2 HBV- and 2 TAA-derived HLA peptides were detected in samples in which no identification was made using unbiased MS. Finally, immunogenicity was demonstrated for 5 HBV-derived and 3 TAA-derived peptides. Conclusions: We present a first HLA-I immunopeptidome of isolated primary human hepatocytes, devoid of immune cells. Identified HBV-derived and TAA-derived peptides directly aid development of antigen-specific immunotherapy for chronic HBV infection and HCC. The described methodology can also be applied to personalise immunotherapeutic treatment of liver diseases in general. Lay summary: Immunotherapy that aims to induce immune responses against a virus or tumour is a promising novel treatment option to treat chronic HBV infection and liver cancer. For the design of successful therapy, it is essential to know which fragments (i.e. peptides) of virus-derived and tumour-specific proteins are presented to the T cells of the immune system by diseased liver cells and are thus good targets for immunotherapy. Here, we have isolated liver cells from patients who have chronic HBV infection and/or liver cancer, analysed what peptides are presented by these cells, and assessed which peptides are able to drive immune responses. |
| first_indexed | 2024-04-12T11:45:45Z |
| format | Article |
| id | doaj.art-0c287e8b13e0403da76d048046b18aa2 |
| institution | Directory Open Access Journal |
| issn | 2589-5559 |
| language | English |
| last_indexed | 2024-04-12T11:45:45Z |
| publishDate | 2022-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | JHEP Reports |
| spelling | doaj.art-0c287e8b13e0403da76d048046b18aa22022-12-22T03:34:22ZengElsevierJHEP Reports2589-55592022-11-01411100576Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinomaMonique T.A. de Beijer0Karel Bezstarosti1Robbie Luijten2Wouter A.S. Doff3Patrick P.C. Boor4Roel F.A. Pieterman5Rachid Bouzid6Paula J. Biesta7Jan N.M. Ijzermans8Michail Doukas9Robert A. de Man10Andrea M. Woltman11Jeroen A.A. Demmers12Sonja I. Buschow13Department of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsProteomics Center, Erasmus MC, Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsProteomics Center, Erasmus MC, Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsDepartment of Surgery, Erasmus MC, Rotterdam, the NetherlandsDepartment of Pathology, Erasmus MC, Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the NetherlandsProteomics Center, Erasmus MC, Rotterdam, the NetherlandsDepartment of Gastroenterology and Hepatology, Erasmus MC University Medical Center Rotterdam, the Netherlands; Corresponding author. Address: Department of Gastroenterology and Hepatology Erasmus MC, Doctor Molewaterplein 40, 3015 GD Rotterdam, the Netherlands.Background & Aims: Antigen-specific immunotherapy is a promising strategy to treat HBV infection and hepatocellular carcinoma (HCC). To facilitate killing of malignant and/or infected hepatocytes, it is vital to know which T cell targets are presented by human leucocyte antigen (HLA)-I complexes on patient-derived hepatocytes. Here, we aimed to reveal the hepatocyte-specific HLA-I peptidome with emphasis on peptides derived from HBV proteins and tumour-associated antigens (TAA) to guide development of antigen-specific immunotherapy. Methods: Primary human hepatocytes were isolated with high purity from (HBV-infected) non-tumour and HCC tissues using a newly designed perfusion-free procedure. Hepatocyte-derived HLA-bound peptides were identified by unbiased mass spectrometry (MS), after which source proteins were subjected to Gene Ontology and pathway analysis. HBV antigen and TAA-derived HLA peptides were searched for using targeted MS, and a selection of peptides was tested for immunogenicity. Results: Using unbiased data-dependent acquisition (DDA), we acquired a high-quality HLA-I peptidome of 2 × 105 peptides that contained 8 HBV-derived peptides and 14 peptides from 8 known HCC-associated TAA that were exclusive to tumours. Of these, 3 HBV- and 12 TAA-derived HLA peptides were detected by targeted MS in the sample they were originally identified in by DDA. Moreover, 2 HBV- and 2 TAA-derived HLA peptides were detected in samples in which no identification was made using unbiased MS. Finally, immunogenicity was demonstrated for 5 HBV-derived and 3 TAA-derived peptides. Conclusions: We present a first HLA-I immunopeptidome of isolated primary human hepatocytes, devoid of immune cells. Identified HBV-derived and TAA-derived peptides directly aid development of antigen-specific immunotherapy for chronic HBV infection and HCC. The described methodology can also be applied to personalise immunotherapeutic treatment of liver diseases in general. Lay summary: Immunotherapy that aims to induce immune responses against a virus or tumour is a promising novel treatment option to treat chronic HBV infection and liver cancer. For the design of successful therapy, it is essential to know which fragments (i.e. peptides) of virus-derived and tumour-specific proteins are presented to the T cells of the immune system by diseased liver cells and are thus good targets for immunotherapy. Here, we have isolated liver cells from patients who have chronic HBV infection and/or liver cancer, analysed what peptides are presented by these cells, and assessed which peptides are able to drive immune responses.http://www.sciencedirect.com/science/article/pii/S2589555922001483Antigen presentationLiver cancerT cell epitopeHLAMHCPeptidome |
| spellingShingle | Monique T.A. de Beijer Karel Bezstarosti Robbie Luijten Wouter A.S. Doff Patrick P.C. Boor Roel F.A. Pieterman Rachid Bouzid Paula J. Biesta Jan N.M. Ijzermans Michail Doukas Robert A. de Man Andrea M. Woltman Jeroen A.A. Demmers Sonja I. Buschow Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma JHEP Reports Antigen presentation Liver cancer T cell epitope HLA MHC Peptidome |
| title | Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma |
| title_full | Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma |
| title_fullStr | Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma |
| title_full_unstemmed | Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma |
| title_short | Immunopeptidome of hepatocytes isolated from patients with HBV infection and hepatocellular carcinoma |
| title_sort | immunopeptidome of hepatocytes isolated from patients with hbv infection and hepatocellular carcinoma |
| topic | Antigen presentation Liver cancer T cell epitope HLA MHC Peptidome |
| url | http://www.sciencedirect.com/science/article/pii/S2589555922001483 |
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