Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments
Recently, long synthetic peptides or in silico-predicted epitope peptides have been used to identify T cell epitopes, but these approaches may not be suitable for investigating naturally processed epitopes. Here, mRNAs, including fragments or predicted epitope sequences of HCMV pp65 antigen, were ge...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/10/5/787 |
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| author | Hong-Seon Pyo Cheol-Hwa Hong Haeyoun Choi In-Cheol Baek Tai-Gyu Kim |
| author_facet | Hong-Seon Pyo Cheol-Hwa Hong Haeyoun Choi In-Cheol Baek Tai-Gyu Kim |
| author_sort | Hong-Seon Pyo |
| collection | DOAJ |
| description | Recently, long synthetic peptides or in silico-predicted epitope peptides have been used to identify T cell epitopes, but these approaches may not be suitable for investigating naturally processed epitopes. Here, mRNAs, including fragments or predicted epitope sequences of HCMV pp65 antigen, were generated by in vitro transcription following transcriptionally active PCR. Then, artificial antigen-presenting cells (aAPCs) expressing a single HLA allotype were transfected with mRNAs to identify epitopes in donors with T cell responses that recognize pp65 antigen restricted to HLA-A*02:01, -A*02:06, or -B*07:02. T cells restricted to a particular HLA allotype showed positive responses in some of the 10 fragment antigens. Among predicted epitopes within these positive fragments, three epitopes of HLA-A*02:01, -A*02:06, and -B*07:02 were confirmed. In addition, T cells expanded by anti-CD3 stimulation for two weeks could also be effectively used for the identification of these T cell epitopes, although there were individual differences. These results demonstrated that fragment antigens and epitopes can be rapidly generated using mRNA, and naturally processed antigenic regions can be detected using aAPCs without a T cell cloning procedure. This method will help to identify novel T cell epitopes for developing immunotherapy and vaccines against infectious diseases and cancer. |
| first_indexed | 2024-03-10T01:39:17Z |
| format | Article |
| id | doaj.art-50378031de3b46c7b253ac69acb4b761 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T01:39:17Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-50378031de3b46c7b253ac69acb4b7612023-11-23T13:27:16ZengMDPI AGVaccines2076-393X2022-05-0110578710.3390/vaccines10050787Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen FragmentsHong-Seon Pyo0Cheol-Hwa Hong1Haeyoun Choi2In-Cheol Baek3Tai-Gyu Kim4Department of Microbiology, College of Medicine, Catholic University of Korea, Seoul 06591, KoreaDepartment of Microbiology, College of Medicine, Catholic University of Korea, Seoul 06591, KoreaDepartment of Microbiology, College of Medicine, Catholic University of Korea, Seoul 06591, KoreaCatholic Hematopoietic Stem Cell Bank, College of Medicine, Catholic University of Korea, Seoul 06591, KoreaDepartment of Microbiology, College of Medicine, Catholic University of Korea, Seoul 06591, KoreaRecently, long synthetic peptides or in silico-predicted epitope peptides have been used to identify T cell epitopes, but these approaches may not be suitable for investigating naturally processed epitopes. Here, mRNAs, including fragments or predicted epitope sequences of HCMV pp65 antigen, were generated by in vitro transcription following transcriptionally active PCR. Then, artificial antigen-presenting cells (aAPCs) expressing a single HLA allotype were transfected with mRNAs to identify epitopes in donors with T cell responses that recognize pp65 antigen restricted to HLA-A*02:01, -A*02:06, or -B*07:02. T cells restricted to a particular HLA allotype showed positive responses in some of the 10 fragment antigens. Among predicted epitopes within these positive fragments, three epitopes of HLA-A*02:01, -A*02:06, and -B*07:02 were confirmed. In addition, T cells expanded by anti-CD3 stimulation for two weeks could also be effectively used for the identification of these T cell epitopes, although there were individual differences. These results demonstrated that fragment antigens and epitopes can be rapidly generated using mRNA, and naturally processed antigenic regions can be detected using aAPCs without a T cell cloning procedure. This method will help to identify novel T cell epitopes for developing immunotherapy and vaccines against infectious diseases and cancer.https://www.mdpi.com/2076-393X/10/5/787epitope identification methodHLA class Iartificial antigen-presenting cellT cell epitopetranscriptionally active PCRmRNA |
| spellingShingle | Hong-Seon Pyo Cheol-Hwa Hong Haeyoun Choi In-Cheol Baek Tai-Gyu Kim Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments Vaccines epitope identification method HLA class I artificial antigen-presenting cell T cell epitope transcriptionally active PCR mRNA |
| title | Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments |
| title_full | Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments |
| title_fullStr | Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments |
| title_full_unstemmed | Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments |
| title_short | Identification of Naturally Processed Epitope Region Using Artificial APC Expressing a Single HLA Class I Allotype and mRNA of HCMV pp65 Antigen Fragments |
| title_sort | identification of naturally processed epitope region using artificial apc expressing a single hla class i allotype and mrna of hcmv pp65 antigen fragments |
| topic | epitope identification method HLA class I artificial antigen-presenting cell T cell epitope transcriptionally active PCR mRNA |
| url | https://www.mdpi.com/2076-393X/10/5/787 |
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