Infection with high-risk types of human papillomavirus (HPV) can cause the development of malignant tumors. To study mechanisms responsible for immune escape of tumor cells infected with HPV16, we previously used mouse oncogenic TC-1 cells producing HPV16 E6 and E7 oncoproteins to derive TC-1 clones resistant to immunization against E7. We have found immunoresistance of the clones to correlate with the point mutation in the E7 oncogene, which resulted in the N53S substitution in the immunodominant epitope RAHYNIVTF (aa 49-57). Here, we have shown that this mutation reduced stabilization of H-2D(b) molecules on RMA-S cells and eliminated immunogenicity of E7. The resistance of TC-1 clones was E7-specific as immunization against E6 inhibited tumor growth. Transduction of the TC-1/F9 clone carrying the mutated epitope with the wild-type E7 gene restored susceptibility to immunization against E7. Our results suggest that mutagenesis of tumor antigens can lead to the escape of malignant cells and should be considered in the development and evaluation of cancer immunotherapy.

Department of Experimental Virology Institute of Hematology and Blood Transfusion U Nemocnice 1 128 20 Prague 2 Czech Republic

Zobrazit více v PubMed

Ahmad M, Rees RC, Ali SA. Escape from immunotherapy: possible mechanisms that influence tumor regression/progression. Cancer Immunol Immunother. 2004;53:844–854. doi: 10.1007/s00262-004-0540-x. PubMed DOI PMC

Bai XF, Liu J, Li O, Zheng P, Liu Y. Antigenic drift as a mechanism for tumor evasion of destruction by cytolytic T lymphocytes. J Clin Invest. 2003;111:1487–1496. PubMed PMC

Bauer S, Heeg K, Wagner H, Lipford GB. Identification of H-2Kb binding and immunogenic peptides from human papilloma virus tumour antigens E6 and E7. Scand J Immunol. 1995;42:317–323. doi: 10.1111/j.1365-3083.1995.tb03662.x. PubMed DOI

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–254. doi: 10.1016/0003-2697(76)90527-3. PubMed DOI

Chen CA, Okayama H. Calcium phosphate-mediated gene transfer: a highly efficient transfection system for stably transforming cells with plasmid DNA. Biotechniques. 1988;6:632–638. PubMed

Chen L, Mizuno MT, Singhal MC, Hu SL, Galloway DA, Hellstrom I, Hellstrom KE. Induction of cytotoxic T lymphocytes specific for a syngeneic tumor expressing the E6 oncoprotein of human papillomavirus type 16. J Immunol. 1992;148:2617–2621. PubMed

Chen LP, Thomas EK, Hu SL, Hellstrom I, Hellstrom KE. Human papillomavirus type 16 nucleoprotein E7 is a tumor rejection antigen. Proc Natl Acad Sci USA. 1991;88:110–114. doi: 10.1073/pnas.88.1.110. PubMed DOI PMC

DuBridge RB, Tang P, Hsia HC, Leong PM, Miller JH, Calos MP. Analysis of mutation in human cells by using an Epstein-Barr virus shuttle system. Mol Cell Biol. 1987;7:379–387. PubMed PMC

Ellis JR, Keating PJ, Baird J, Hounsell EF, Renouf DV, Rowe M, Hopkins D, Duggan-Keen MF, Bartholomew JS, Young LS. The association of an HPV16 oncogene variant with HLA-B7 has implications for vaccine design in cervical cancer. Nat Med. 1995;1:464–470. doi: 10.1038/nm0595-464. PubMed DOI

Feltkamp MC, Smits HL, Vierboom MP, Minnaar RP, de Jongh BM, Drijfhout JW, ter Schegget J, Melief CJ, Kast WM. Vaccination with cytotoxic T lymphocyte epitope-containing peptide protects against a tumor induced by human papillomavirus type 16-transformed cells. Eur J Immunol. 1993;23:2242–2249. doi: 10.1002/eji.1830230929. PubMed DOI

He Z, Wlazlo AP, Kowalczyk DW, Cheng J, Xiang ZQ, Giles-Davis W, Ertl HC. Viral recombinant vaccines to the E6 and E7 antigens of HPV-16. Virology. 2000;270:146–161. doi: 10.1006/viro.2000.0271. PubMed DOI

Hicklin DJ, Marincola FM, Ferrone S. HLA class I antigen downregulation in human cancers: T-cell immunotherapy revives an old story. Mol Med Today. 1999;5:178–186. doi: 10.1016/S1357-4310(99)01451-3. PubMed DOI

Jainchill JL, Aaronson SA, Todaro GJ. Murine sarcoma and leukemia viruses: assay using clonal lines of contact-inhibited mouse cells. J Virol. 1969;4:549–553. PubMed PMC

Karre K, Ljunggren HG, Piontek G, Kiessling R. Selective rejection of H-2-deficient lymphoma variants suggests alternative immune defence strategy. Nature. 1986;319:675–678. doi: 10.1038/319675a0. PubMed DOI

Keating PJ, Cromme FV, Duggan KM, Snijders PJ, Walboomers JM, Hunter RD, Dyer PA, Stern PL. Frequency of down-regulation of individual HLA-A and -B alleles in cervical carcinomas in relation to TAP-1 expression. Br J Cancer. 1995;72:405–411. PubMed PMC

Khammanivong V, Liu XS, Liu WJ, Rodda SJ, Leggatt GR, Tindle RW, Frazer IH, Fernando GJ. Paucity of functional CTL epitopes in the E7 oncoprotein of cervical cancer associated human papillomavirus type 16. Immunol Cell Biol. 2003;81:1–7. doi: 10.1046/j.1440-1711.2003.01130.x. PubMed DOI

Koopman LA, Corver WE, van der Slik AR, Giphart MJ, Fleuren GJ. Multiple genetic alterations cause frequent and heterogeneous human histocompatibility leukocyte antigen class I loss in cervical cancer. J Exp Med. 2000;191:961–976. doi: 10.1084/jem.191.6.961. PubMed DOI PMC

Lee KH, Panelli MC, Kim CJ, Riker AI, Bettinotti MP, Roden MM, Fetsch P, Abati A, Rosenberg SA, Marincola FM. Functional dissociation between local and systemic immune response during anti-melanoma peptide vaccination. J Immunol. 1998;161:4183–4194. PubMed

Lichtig H, Algrisi M, Botzer LE, Abadi T, Verbitzky Y, Jackman A, Tommasino M, Zehbe I, Sherman L. HPV16 E6 natural variants exhibit different activities in functional assays relevant to the carcinogenic potential of E6. Virology. 2006;350:216–227. doi: 10.1016/j.virol.2006.01.038. PubMed DOI

Lin KY, Guarnieri FG, Staveley OK, Levitsky HI, August JT, Pardoll DM, Wu TC. Treatment of established tumors with a novel vaccine that enhances major histocompatibility class II presentation of tumor antigen. Cancer Res. 1996;56:21–26. PubMed

Lipford GB, Bauer S, Wagner H, Heeg K. Peptide engineering allows cytotoxic T-cell vaccination against human papilloma virus tumour antigen, E6. Immunology. 1995;84:298–303. PubMed PMC

Ljunggren HG, Stam NJ, Ohlen C, Neefjes JJ, Hoglund P, Heemels MT, Bastin J, Schumacher TN, Townsend A, Karre K. Empty MHC class I molecules come out in the cold. Nature. 1990;346:476–480. doi: 10.1038/346476a0. PubMed DOI

Londesborough P, Ho L, Terry G, Cuzick J, Wheeler C, Singer A. Human papillomavirus genotype as a predictor of persistence and development of high-grade lesions in women with minor cervical abnormalities. Int J Cancer. 1996;69:364–368. doi: 10.1002/(SICI)1097-0215(19961021)69:5<364::AID-IJC2>3.0.CO;2-3. PubMed DOI

Munger K, Howley PM. Human papillomavirus immortalization and transformation functions. Virus Res. 2002;89:213–228. doi: 10.1016/S0168-1702(02)00190-9. PubMed DOI

Nasiell K, Roger V, Nasiell M. Behavior of mild cervical dysplasia during long-term follow-up. Obstet Gynecol. 1986;67:665–669. doi: 10.1097/00006250-198605000-00012. PubMed DOI

Nemeckova S, Stranska R, Subrtova J, Kutinova L, Otahal P, Hainz P, Maresova L, Sroller V, Hamsikova E, Vonka V. Immune response to E7 protein of human papillomavirus type 16 anchored on the cell surface. Cancer Immunol Immunother. 2002;51:111–119. doi: 10.1007/s00262-001-0261-3. PubMed DOI PMC

Pawelec G, Heinzel S, Kiessling R, Muller L, Ouyang Q, Zeuthen J. Escape mechanisms in tumor immunity: a year 2000 update. Crit Rev Oncog. 2000;11:97–133. PubMed

Peng S, Ji H, Trimble C, He L, Tsai YC, Yeatermeyer J, Boyd DA, Hung CF, Wu TC. Development of a DNA vaccine targeting human papillomavirus type 16 oncoprotein E6. J Virol. 2004;78:8468–8476. doi: 10.1128/JVI.78.16.8468-8476.2004. PubMed DOI PMC

Reche PA, Glutting JP, Reinherz EL. Prediction of MHC class I binding peptides using profile motifs. Hum Immunol. 2002;63:701–709. doi: 10.1016/S0198-8859(02)00432-9. PubMed DOI

Reche PA, Glutting JP, Zhang H, Reinherz EL. Enhancement to the RANKPEP resource for the prediction of peptide binding to MHC molecules using profiles. Immunogenetics. 2004;56:405–419. doi: 10.1007/s00251-004-0709-7. PubMed DOI

Riker A, Cormier J, Panelli M, Kammula U, Wang E, Abati A, Fetsch P, Lee KH, Steinberg S, Rosenberg S, Marincola F. Immune selection after antigen-specific immunotherapy of melanoma. Surgery. 1999;126:112–120. PubMed

Sadovnikova E, Zhu X, Collins SM, Zhou J, Vousden K, Crawford L, Beverley P, Stauss HJ. Limitations of predictive motifs revealed by cytotoxic T lymphocyte epitope mapping of the human papilloma virus E7 protein. Int Immunol. 1994;6:289–296. doi: 10.1093/intimm/6.2.289. PubMed DOI

Scott M, Nakagawa M, Moscicki AB. Cell-mediated immune response to human papillomavirus infection. Clin Diagn Lab Immunol. 2001;8:209–220. doi: 10.1128/CDLI.8.2.209-220.2001. PubMed DOI PMC

Seliger B, Maeurer MJ, Ferrone S. Antigen-processing machinery breakdown and tumor growth. Immunol Today. 2000;21:455–464. doi: 10.1016/S0167-5699(00)01692-3. PubMed DOI

Smahel M, Pokorna D, Mackova J, Vlasak J. Enhancement of immunogenicity of HPV16 E7 oncogene by fusion with E. coli beta-glucuronidase. J Gene Med. 2004;6:1092–1101. doi: 10.1002/jgm.596. PubMed DOI

Smahel M, Sima P, Ludvikova V, Marinov I, Pokorna D, Vonka V. Immunisation with modified HPV16 E7 genes against mouse oncogenic TC-1 cell sublines with downregulated expression of MHC class I molecules. Vaccine. 2003;21:1125–1136. doi: 10.1016/S0264-410X(02)00519-4. PubMed DOI

Smahel M, Sima P, Ludvikova V, Vonka V. Modified HPV16 E7 genes as DNA vaccine against E7-containing oncogenic cells. Virology. 2001;281:231–238. doi: 10.1006/viro.2000.0794. PubMed DOI

Smahel M, Smahelova J, Tejklova P, Tachezy R, Marinov I. Characterization of cell lines derived from tumors induced by TC-1 cells in mice preimmunized against HPV16 E7 oncoprotein. Int J Oncol. 2005;27:731–742. PubMed

Zehbe I, Mytilineos J, Wikstrom I, Henriksen R, Edler L, Tommasino M. Association between human papillomavirus 16 E6 variants and human leukocyte antigen class I polymorphism in cervical cancer of Swedish women. Hum Immunol. 2003;64:538–542. doi: 10.1016/S0198-8859(03)00033-8. PubMed DOI

zur Hausen H. Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer. 2002;2:342–350. doi: 10.1038/nrc798. PubMed DOI

Chemokine binding protein vCCI attenuates vaccinia virus without affecting the cellular response elicited by immunization with a recombinant vaccinia vector carrying the HPV16 E7 gene