PMID- 36224625 OWN - NLM STAT- MEDLINE DCOM- 20221014 LR - 20230120 IS - 1423-0127 (Electronic) IS - 1021-7770 (Print) IS - 1021-7770 (Linking) VI - 29 IP - 1 DP - 2022 Oct 12 TI - Identification of human MHC-I HPV18 E6/E7-specific CD8 + T cell epitopes and generation of an HPV18 E6/E7-expressing adenosquamous carcinoma in HLA-A2 transgenic mice. PG - 80 LID - 10.1186/s12929-022-00864-5 [doi] LID - 80 AB - BACKGROUND: Human Papillomavirus type 18 (HPV18) is a high-risk HPV that is commonly associated with cervical cancer. HPV18 oncogenes E6 and E7 are associated with the malignant transformation of cells, thus the identification of human leukocyte antigen (HLA)-restricted E6/E7 peptide-specific CD8 + T cell epitopes and the creation of a HPV18 E6/E7 expressing cervicovaginal tumor in HLA-A2 transgenic mice will be significant for vaccine development. METHODS: In the below study, we characterized various human HLA class I-restricted HPV18 E6 and E7-specific CD8 + T cells mediated immune responses in HLA class I transgenic mice using DNA vaccines encoding HPV18E6 and HPV18E7. We then confirmed HLA-restricted E6/E7 specific CD8 + T cell epitopes using splenocytes from vaccinated mice stimulated with HPV18E6/E7 peptides. Furthermore, we used oncogenic DNA plasmids encoding HPV18E7E6(delD70), luciferase, cMyc, and AKT to create a spontaneous cervicovaginal carcinoma model in HLA-A2 transgenic mice. RESULTS: Therapeutic HPV18 E7 DNA vaccination did not elicit any significant CD8 + T cell response in HLA-A1, HLA-24, HLA-B7, HLA-B44 transgenic or wild type C57BL/6 mice, but it did generate a strong HLA-A2 and HLA-A11 restricted HPV18E7-specific CD8 + T cell immune response. We found that a single deletion of aspartic acid (D) at location 70 in HPV18E6 DNA abolishes the presentation of HPV18 E6 peptide (aa67-75) by murine MHC class I. We found that the DNA vaccine with this mutant HPV18 E6 generated E6-specific CD8 + T cells in HLA-A2. HLA-A11, HLA-A24 and HLA-b40 transgenic mice. Of note, HLA-A2 restricted, HPV18 E7 peptide (aa7-15)- and HPV18 E6 peptide (aa97-105)-specific epitopes are endogenously processed by HPV18 positive Hela-AAD (HLA-A(*)0201/D(d)) cells. Finally, we found that injection of DNA plasmids encoding HPV18E7E6(delD70), AKT, cMyc, and SB100 can result in the development of adenosquamous carcinoma in the cervicovaginal tract of HLA-A2 transgenic mice. CONCLUSIONS: We characterized various human HLA class I-restricted HPV18 E6/E7 peptide specific CD8 + T cell epitopes in human HLA class I transgenic mice. We demonstrated that HPV18 positive Hela cells expressing chimeric HLA-A2 (AAD) do present both HLA-A2-restricted HPV18 E7 (aa7-15)- and HPV18 E6 (aa97-105)-specific CD8 + T cell epitopes. A mutant HPV18E6 that had a single deletion at location 70 obliterates the E6 presentation by murine MHC class I and remains oncogenic. The identification of these human MHC restricted HPV antigen specific epitopes as well as the HPV18E6/E7 expressing adenosquamous cell carcinoma model may have significant future translational potential. CI - (c) 2022. The Author(s). FAU - Peng, Shiwen AU - Peng S AD - Department of Pathology, The Johns Hopkins University, Baltimore, MD, USA. FAU - Xing, Deyin AU - Xing D AD - Department of Pathology, The Johns Hopkins University, Baltimore, MD, USA. FAU - Ferrall, Louise AU - Ferrall L AD - Department of Pathology, The Johns Hopkins University, Baltimore, MD, USA. FAU - Tsai, Ya-Chea AU - Tsai YC AD - Department of Pathology, The Johns Hopkins University, Baltimore, MD, USA. FAU - Hung, Chien-Fu AU - Hung CF AD - Department of Pathology, The Johns Hopkins University, Baltimore, MD, USA. chung2@jhmi.edu. AD - Department of Oncology, The Johns Hopkins University, Baltimore, MD, USA. chung2@jhmi.edu. AD - Department of Obstetrics and Gynecology, The Johns Hopkins University, Baltimore, MD, USA. chung2@jhmi.edu. AD - The Johns Hopkins Medical Institutions, CRB II Room 307, 1550 Orleans St., Baltimore, MD, 21231, USA. chung2@jhmi.edu. FAU - Wu, T-C AU - Wu TC AUID- ORCID: 0000-0003-1623-4584 AD - Department of Pathology, The Johns Hopkins University, Baltimore, MD, USA. wutc@jhmi.edu. AD - Department of Oncology, The Johns Hopkins University, Baltimore, MD, USA. wutc@jhmi.edu. AD - Department of Obstetrics and Gynecology, The Johns Hopkins University, Baltimore, MD, USA. wutc@jhmi.edu. AD - Department of Molecular Microbiology and Immunology, The Johns Hopkins University, Baltimore, MD, USA. wutc@jhmi.edu. AD - The Johns Hopkins Medical Institutions, CRB II Room 309, 1550 Orleans St., Baltimore, MD, 21231, USA. wutc@jhmi.edu. LA - eng GR - P50 CA098252/CA/NCI NIH HHS/United States GR - R01 CA237067/CA/NCI NIH HHS/United States GR - P50CA098252/CA/NCI NIH HHS/United States GR - R50 CA251953/CA/NCI NIH HHS/United States GR - R50 CA251953-02/CA/NCI NIH HHS/United States GR - R01CA237067/CA/NCI NIH HHS/United States PT - Journal Article DEP - 20221012 PL - England TA - J Biomed Sci JT - Journal of biomedical science JID - 9421567 RN - 0 (Epitopes, T-Lymphocyte) RN - 0 (HLA-A Antigens) RN - 0 (HLA-A1 Antigen) RN - 0 (HLA-A11 Antigen) RN - 0 (HLA-A2 Antigen) RN - 0 (HLA-A24 Antigen) RN - 0 (HLA-B40 Antigen) RN - 0 (HLA-B44 Antigen) RN - 0 (HLA-B7 Antigen) RN - 0 (Oncogene Proteins, Viral) RN - 0 (Peptides) RN - 0 (Vaccines, DNA) RN - 30KYC7MIAI (Aspartic Acid) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) SB - IM MH - Animals MH - Aspartic Acid MH - CD8-Positive T-Lymphocytes MH - *Carcinoma, Adenosquamous/complications MH - Epitopes, T-Lymphocyte/genetics MH - Female MH - HLA-A Antigens MH - HLA-A1 Antigen MH - HLA-A11 Antigen MH - HLA-A2 Antigen/genetics MH - HLA-A24 Antigen MH - HLA-B40 Antigen MH - HLA-B44 Antigen MH - HLA-B7 Antigen MH - HeLa Cells MH - Human papillomavirus 18 MH - Humans MH - Mice MH - Mice, Inbred C57BL MH - Mice, Transgenic MH - *Oncogene Proteins, Viral/genetics MH - *Papillomavirus Infections/complications MH - Peptides MH - Proto-Oncogene Proteins c-akt MH - T-Lymphocytes, Cytotoxic MH - *Vaccines, DNA/genetics PMC - PMC9554842 OTO - NOTNLM OT - Adenosquamous cell carcinoma OT - E6 OT - E7 OT - Epitope OT - HLA-A2 OT - HPV18 OT - Mouse model COIS- T.-C. Wu is a cofounders of and has an equity ownership interest in Papivax LLC. Also, he owns Papivax Biotech, Inc., stock and is a member of the Scientific Advisory Board of Papivax Biotech, Inc. No other author declares a competing interest. EDAT- 2022/10/13 06:00 MHDA- 2022/10/15 06:00 PMCR- 2022/10/12 CRDT- 2022/10/12 23:46 PHST- 2022/02/15 00:00 [received] PHST- 2022/09/28 00:00 [accepted] PHST- 2022/10/12 23:46 [entrez] PHST- 2022/10/13 06:00 [pubmed] PHST- 2022/10/15 06:00 [medline] PHST- 2022/10/12 00:00 [pmc-release] AID - 10.1186/s12929-022-00864-5 [pii] AID - 864 [pii] AID - 10.1186/s12929-022-00864-5 [doi] PST - epublish SO - J Biomed Sci. 2022 Oct 12;29(1):80. doi: 10.1186/s12929-022-00864-5.