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. 1993 May;67(5):2799–2806. doi: 10.1128/jvi.67.5.2799-2806.1993

Production and characterization of human proliferative T-cell clones specific for human papillomavirus type 1 E4 protein.

J C Steele 1, T Stankovic 1, P H Gallimore 1
PMCID: PMC237604  PMID: 7682626

Abstract

Human papillomavirus type 1 (HPV1) virions and E4 protein purified from cutaneous warts were tested in lymphocyte proliferation assays using normal individuals. Both antigens were found to be capable of eliciting good lymphoproliferative responses. Several T-cell clones specific for wart E4 protein were obtained from a donor who had consistently responded very well to E4 in these initial assays. They were maintained in culture by repeated stimulation with antigen and interleukin-2, using an autologous mitomycin-treated lymphoblastoid cell line as a source of antigen-presenting cells. Two of these clones (3F5 and 4A8), which behaved identically, have been studied in more detail. A series of overlapping synthetic peptides covering the entire E1 E4 protein sequence was used to identify a single T-cell epitope which maps to a strongly hydrophilic region spanning amino acid residues 38 to 50. We have also tested the ability of a panel of major histocompatibility complex class II-matched and -mismatched lymphoblastoid cell lines to present this peptide to the T-cell clones in proliferation assays. The study reports that the epitope is restricted through HLA-DQ7 and that it can be recognized by T cells with different T-cell receptor gene rearrangements.

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Selected References

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  1. Altmann A., Jochmus-Kudielka I., Frank R., Gausepohl H., Moebius U., Gissmann L., Meuer S. C. Definition of immunogenic determinants of the human papillomavirus type 16 nucleoprotein E7. Eur J Cancer. 1992;28(2-3):326–333. doi: 10.1016/s0959-8049(05)80047-4. [DOI] [PubMed] [Google Scholar]
  2. Berman A., Winkelmann R. K. Involuting common warts. Clinical and histopathologic findings. J Am Acad Dermatol. 1980 Oct;3(4):356–362. doi: 10.1016/s0190-9622(80)80328-8. [DOI] [PubMed] [Google Scholar]
  3. Bjorkman P. J., Saper M. A., Samraoui B., Bennett W. S., Strominger J. L., Wiley D. C. Structure of the human class I histocompatibility antigen, HLA-A2. Nature. 1987 Oct 8;329(6139):506–512. doi: 10.1038/329506a0. [DOI] [PubMed] [Google Scholar]
  4. Bjorkman P. J., Saper M. A., Samraoui B., Bennett W. S., Strominger J. L., Wiley D. C. The foreign antigen binding site and T cell recognition regions of class I histocompatibility antigens. Nature. 1987 Oct 8;329(6139):512–518. doi: 10.1038/329512a0. [DOI] [PubMed] [Google Scholar]
  5. Bunney M. H. Viral warts: a new look at an old problem. Br Med J (Clin Res Ed) 1986 Oct 25;293(6554):1045–1047. doi: 10.1136/bmj.293.6554.1045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chardonnet Y., Viac J., Staquet M. J., Thivolet J. Cell-mediated immunity to human papillomavirus. Clin Dermatol. 1985 Oct-Dec;3(4):156–164. doi: 10.1016/0738-081x(85)90060-4. [DOI] [PubMed] [Google Scholar]
  7. Chen L. P., Thomas E. K., Hu S. L., Hellström I., Hellström K. E. Human papillomavirus type 16 nucleoprotein E7 is a tumor rejection antigen. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):110–114. doi: 10.1073/pnas.88.1.110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chicz R. M., Urban R. G., Lane W. S., Gorga J. C., Stern L. J., Vignali D. A., Strominger J. L. Predominant naturally processed peptides bound to HLA-DR1 are derived from MHC-related molecules and are heterogeneous in size. Nature. 1992 Aug 27;358(6389):764–768. doi: 10.1038/358764a0. [DOI] [PubMed] [Google Scholar]
  9. Comerford S. A., McCance D. J., Dougan G., Tite J. P. Identification of T- and B-cell epitopes of the E7 protein of human papillomavirus type 16. J Virol. 1991 Sep;65(9):4681–4690. doi: 10.1128/jvi.65.9.4681-4690.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cubie H. A., Norval M., Crawford L., Banks L., Crook T. Lymphoproliferative response to fusion proteins of human papillomaviruses in patients with cervical intraepithelial neoplasia. Epidemiol Infect. 1989 Dec;103(3):625–632. doi: 10.1017/s0950268800031022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cubie H. A., Norval M. Humoral and cellular immunity to papillomavirus in patients with cervical dysplasia. J Med Virol. 1988 Jan;24(1):85–95. doi: 10.1002/jmv.1890240111. [DOI] [PubMed] [Google Scholar]
  12. Davies D. H., Hill C. M., Rothbard J. B., Chain B. M. Definition of murine T helper cell determinants in the major capsid protein of human papillomavirus type 16. J Gen Virol. 1990 Nov;71(Pt 11):2691–2698. doi: 10.1099/0022-1317-71-11-2691. [DOI] [PubMed] [Google Scholar]
  13. DeLisi C., Berzofsky J. A. T-cell antigenic sites tend to be amphipathic structures. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7048–7052. doi: 10.1073/pnas.82.20.7048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Doorbar J., Campbell D., Grand R. J., Gallimore P. H. Identification of the human papilloma virus-1a E4 gene products. EMBO J. 1986 Feb;5(2):355–362. doi: 10.1002/j.1460-2075.1986.tb04219.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Doorbar J., Evans H. S., Coneron I., Crawford L. V., Gallimore P. H. Analysis of HPV-1 E4 gene expression using epitope-defined antibodies. EMBO J. 1988 Mar;7(3):825–833. doi: 10.1002/j.1460-2075.1988.tb02881.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Doorbar J., Gallimore P. H. Identification of proteins encoded by the L1 and L2 open reading frames of human papillomavirus 1a. J Virol. 1987 Sep;61(9):2793–2799. doi: 10.1128/jvi.61.9.2793-2799.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Galloway D. A., Jenison S. A. Characterization of the humoral immune response to genital papillomaviruses. Mol Biol Med. 1990 Feb;7(1):59–72. [PubMed] [Google Scholar]
  18. Gissmann L., Gross G. Association of HPV with human genital tumors. Clin Dermatol. 1985 Oct-Dec;3(4):124–129. doi: 10.1016/0738-081x(85)90056-2. [DOI] [PubMed] [Google Scholar]
  19. Grand R. J., Doorbar J., Smith K. J., Coneron I., Gallimore P. H. Phosphorylation of the human papillomavirus type 1 E4 proteins in vivo and in vitro. Virology. 1989 May;170(1):201–213. doi: 10.1016/0042-6822(89)90367-x. [DOI] [PubMed] [Google Scholar]
  20. Haas W., Kaufman S., Martinez C. The development and function of gamma delta T cells. Immunol Today. 1990 Oct;11(10):340–343. doi: 10.1016/0167-5699(90)90133-t. [DOI] [PubMed] [Google Scholar]
  21. Han R., Breitburd F., Marche P. N., Orth G. Linkage of regression and malignant conversion of rabbit viral papillomas to MHC class II genes. Nature. 1992 Mar 5;356(6364):66–68. doi: 10.1038/356066a0. [DOI] [PubMed] [Google Scholar]
  22. Hunt D. F., Michel H., Dickinson T. A., Shabanowitz J., Cox A. L., Sakaguchi K., Appella E., Grey H. M., Sette A. Peptides presented to the immune system by the murine class II major histocompatibility complex molecule I-Ad. Science. 1992 Jun 26;256(5065):1817–1820. doi: 10.1126/science.1319610. [DOI] [PubMed] [Google Scholar]
  23. Ivanyi L., Morison W. L. In vitro lymphocyte stimulation by wart antigen in man. Br J Dermatol. 1976 May;94(5):523–527. doi: 10.1111/j.1365-2133.1976.tb05140.x. [DOI] [PubMed] [Google Scholar]
  24. Iwatsuki K., Tagami H., Takigawa M., Yamada M. Plane warts under spontaneous regression. Immunopathologic study on cellular constituents leading to the inflammatory reaction. Arch Dermatol. 1986 Jun;122(6):655–659. doi: 10.1001/archderm.122.6.655. [DOI] [PubMed] [Google Scholar]
  25. Jardetzky T. S., Lane W. S., Robinson R. A., Madden D. R., Wiley D. C. Identification of self peptides bound to purified HLA-B27. Nature. 1991 Sep 26;353(6342):326–329. doi: 10.1038/353326a0. [DOI] [PubMed] [Google Scholar]
  26. Jarrett W. F., O'Neil B. W., Gaukroger J. M., Smith K. T., Laird H. M., Campo M. S. Studies on vaccination against papillomaviruses: the immunity after infection and vaccination with bovine papillomaviruses of different types. Vet Rec. 1990 May 12;126(19):473–475. [PubMed] [Google Scholar]
  27. Jarrett W. F., Smith K. T., O'Neil B. W., Gaukroger J. M., Chandrachud L. M., Grindlay G. J., McGarvie G. M., Campo M. S. Studies on vaccination against papillomaviruses: prophylactic and therapeutic vaccination with recombinant structural proteins. Virology. 1991 Sep;184(1):33–42. doi: 10.1016/0042-6822(91)90819-w. [DOI] [PubMed] [Google Scholar]
  28. Kienzler J. L., Lemoine M. T., Orth G., Jibard N., Blanc D., Laurent R., Agache P. Humoral and cell-mediated immunity to human papillomavirus type 1 (HPV-1) in human warts. Br J Dermatol. 1983 Jun;108(6):665–672. doi: 10.1111/j.1365-2133.1983.tb01078.x. [DOI] [PubMed] [Google Scholar]
  29. Lahiri D. K., Nurnberger J. I., Jr A rapid non-enzymatic method for the preparation of HMW DNA from blood for RFLP studies. Nucleic Acids Res. 1991 Oct 11;19(19):5444–5444. doi: 10.1093/nar/19.19.5444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lamb J. R., Eckels D. D., Phelan M., Lake P., Woody J. N. Antigen-specific human T lymphocyte clones: viral antigen specificity of influenza virus-immune clones. J Immunol. 1982 Mar;128(3):1428–1432. [PubMed] [Google Scholar]
  31. Lee A. K., Eisinger M. Cell-mediated immunity (CMI) to human wart virus and wart-associated tissue antigens. Clin Exp Immunol. 1976 Dec;26(3):419–424. [PMC free article] [PubMed] [Google Scholar]
  32. Morison W. L. In vitro assay of immunity to human wart antigen. Br J Dermatol. 1975 Nov;93(5):545–552. doi: 10.1111/j.1365-2133.1975.tb02247.x. [DOI] [PubMed] [Google Scholar]
  33. Pyrhönen S., Jablonska S., Obałek S., Kuismanen E. Immune reactions in epidermodysplasia verruciformis. Br J Dermatol. 1980 Mar;102(3):247–254. doi: 10.1111/j.1365-2133.1980.tb08136.x. [DOI] [PubMed] [Google Scholar]
  34. Rabin H., Hopkins R. F., 3rd, Ruscetti F. W., Neubauer R. H., Brown R. L., Kawakami T. G. Spontaneous release of a factor with properties of T cell growth factor from a continuous line of primate tumor T cells. J Immunol. 1981 Nov;127(5):1852–1856. [PubMed] [Google Scholar]
  35. Rothbard J. B., Taylor W. R. A sequence pattern common to T cell epitopes. EMBO J. 1988 Jan;7(1):93–100. doi: 10.1002/j.1460-2075.1988.tb02787.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rudensky AYu, Preston-Hurlburt P., al-Ramadi B. K., Rothbard J., Janeway C. A., Jr Truncation variants of peptides isolated from MHC class II molecules suggest sequence motifs. Nature. 1992 Oct 1;359(6394):429–431. doi: 10.1038/359429a0. [DOI] [PubMed] [Google Scholar]
  37. Rötzschke O., Falk K., Deres K., Schild H., Norda M., Metzger J., Jung G., Rammensee H. G. Isolation and analysis of naturally processed viral peptides as recognized by cytotoxic T cells. Nature. 1990 Nov 15;348(6298):252–254. doi: 10.1038/348252a0. [DOI] [PubMed] [Google Scholar]
  38. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  39. Schumacher T. N., De Bruijn M. L., Vernie L. N., Kast W. M., Melief C. J., Neefjes J. J., Ploegh H. L. Peptide selection by MHC class I molecules. Nature. 1991 Apr 25;350(6320):703–706. doi: 10.1038/350703a0. [DOI] [PubMed] [Google Scholar]
  40. Shepherd P. S., Tran T. T., Rowe A. J., Cridland J. C., Comerford S. A., Chapman M. G., Rayfield L. S. T cell responses to the human papillomavirus type 16 E7 protein in mice of different haplotypes. J Gen Virol. 1992 May;73(Pt 5):1269–1274. doi: 10.1099/0022-1317-73-5-1269. [DOI] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Stauss H. J., Davies H., Sadovnikova E., Chain B., Horowitz N., Sinclair C. Induction of cytotoxic T lymphocytes with peptides in vitro: identification of candidate T-cell epitopes in human papilloma virus. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7871–7875. doi: 10.1073/pnas.89.17.7871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Steele J. C., Gallimore P. H. Humoral assays of human sera to disrupted and nondisrupted epitopes of human papillomavirus type 1. Virology. 1990 Feb;174(2):388–398. doi: 10.1016/0042-6822(90)90092-6. [DOI] [PubMed] [Google Scholar]
  44. Strang G., Hickling J. K., McIndoe G. A., Howland K., Wilkinson D., Ikeda H., Rothbard J. B. Human T cell responses to human papillomavirus type 16 L1 and E6 synthetic peptides: identification of T cell determinants, HLA-DR restriction and virus type specificity. J Gen Virol. 1990 Feb;71(Pt 2):423–431. doi: 10.1099/0022-1317-71-2-423. [DOI] [PubMed] [Google Scholar]
  45. Tagami H., Oku T., Iwatsuki K. Primary tissue culture of spontaneously regressing flat warts. In vitro attack by mononuclear cells against wart-derived epidermal cells. Cancer. 1985 May 15;55(10):2437–2441. doi: 10.1002/1097-0142(19850515)55:10<2437::aid-cncr2820551023>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  46. Thivolet J., Viac J., Staquet M. J. Cell-mediated immunity in wart infection. Int J Dermatol. 1982 Mar;21(2):94–98. doi: 10.1111/j.1365-4362.1982.tb00510.x. [DOI] [PubMed] [Google Scholar]
  47. Tindle R. W., Fernando G. J., Sterling J. C., Frazer I. H. A "public" T-helper epitope of the E7 transforming protein of human papillomavirus 16 provides cognate help for several E7 B-cell epitopes from cervical cancer-associated human papillomavirus genotypes. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5887–5891. doi: 10.1073/pnas.88.13.5887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Townsend A., Ohlén C., Bastin J., Ljunggren H. G., Foster L., Kärre K. Association of class I major histocompatibility heavy and light chains induced by viral peptides. Nature. 1989 Aug 10;340(6233):443–448. doi: 10.1038/340443a0. [DOI] [PubMed] [Google Scholar]
  49. Van Bleek G. M., Nathenson S. G. Isolation of an endogenously processed immunodominant viral peptide from the class I H-2Kb molecule. Nature. 1990 Nov 15;348(6298):213–216. doi: 10.1038/348213a0. [DOI] [PubMed] [Google Scholar]
  50. Viac J., Thivolet J., Hegazy M. R., Chardonnet Y., Dambuyant C. Comparative study of delayed hypersensitivity skin reactions and antibodies to human papilloma virus (HPV). Clin Exp Immunol. 1977 Aug;29(2):240–246. [PMC free article] [PubMed] [Google Scholar]
  51. Wank R., Thomssen C. High risk of squamous cell carcinoma of the cervix for women with HLA-DQw3. Nature. 1991 Aug 22;352(6337):723–725. doi: 10.1038/352723a0. [DOI] [PubMed] [Google Scholar]
  52. van Bleek G. M., Nathenson S. G. The structure of the antigen-binding groove of major histocompatibility complex class I molecules determines specific selection of self-peptides. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11032–11036. doi: 10.1073/pnas.88.24.11032. [DOI] [PMC free article] [PubMed] [Google Scholar]

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