Abstract
The ethyl-N-nitrosourea-induced rat glioma N32 was treated with the mutagenic compoundN-methyl-N′-nitro-N-nitrosoguanidine and the surviving cells cloned by limiting dilution. Out of 20 clones tested 8 did not produce tumors subcutaneously even after challenge doses 3 log units above the minimal tumor dose for N32. All of 5 clones grew in a retarded manner intracerebrally but produced tumors in some animals. Preimmunizations with three of the rejected clones (tum−) gave protection against subcutaneous and intracerebral isografts of the unmutated N32. This effect could be enhanced if the cells used for immunizations were pretreated with interferon γ (IFNγ) for 48 h. If immunizations were started subsequent to challenge, only immunization with one of two tested tum− clones pretreated with IFNγ induced significant rejection against intracerebral N32 isografts. Both N32 and its tum− closes were MHC class I positive and MHC class II negative. IFNγ treatment enhanced the MHC class I expression with 20%–90% on the tum− clones and with 40% on N32. MHC class II expression could be induced on N32 cells after 7 days of IFNγ treatment but not on any of the tum− clones tested. We conclude that the enhancing effect of IFNγ treatment on tumor isograft rejection may depend on up-regulation of MHC class I but not of MHC class II. This investigation demonstrates that it is possible to induce rejection of weakly immunogenic intracerebral brain tumors by immunization with selected highly immunogenic tumor cell mutants. In conjunction with relevant cytokines, the cross-protective effect of these tum− variants might be further enhanced and serve as a model for immunotherapy against malignant human brain tumors.
Key words: IFNγ, Immunization, Immunogenicity, Isograft rejection, Mutagen treatment, Rat brain tumor
References
- 1.Blankenstein T, Rowley D, Schreiber H. Cytokines and cancer: experimental systems. Curr Opin Immunol. 1991;3:694. doi: 10.1016/0952-7915(91)90098-l. [DOI] [PubMed] [Google Scholar]
- 2.Bloom WH, Carstairs KC, Crompton MR, McKissock W. Autologous glioma transplantation. Lancet. 1960;9:77. doi: 10.1016/s0140-6736(60)91219-8. [DOI] [PubMed] [Google Scholar]
- 3.Bloom HJG, Peckham MJ, Richardson AE, Alexander PA, Payne PM. Glioblastoma multiforme: a controlled trial to asses the value of specific active immunotherapy in patients treated by radical surgery and radiotherapy. Br J Cancer. 1973;27:253. doi: 10.1038/bjc.1973.30. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Bodmer S, Siepl C, Fontana A. Neuroimmune networks: Physiology and diseases. New York: Alan R Liss; 1989. Immunoregulatory factors secreted by glioblastoma cells: glioblastoma derived T-cell supressor factor/transforming growth factor-beta; p. 73. [Google Scholar]
- 5.Boon T. Tum− variants: immunogenic variants obtained by mutagen treatment of tumor cells. Immunol Today. 1985;6:307. doi: 10.1016/0167-5699(85)90182-3. [DOI] [PubMed] [Google Scholar]
- 6.Boon T, Van Snick J, Van Pel A, Uyttenhove C, Marchand M. Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815 II. T-lymphocyte mediated cytolysis. J Exp Med. 1980;152:1184. doi: 10.1084/jem.152.5.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Brent L. Pthophysiology of the blood-brain barrier. Amsterdam: Elsevier; 1990. Immunologically privileged sites; p. 383. [Google Scholar]
- 8.Cravioto HM, De Bernardo E, Hochwald GM, Thorbecke JG. Immunity to transplantable nitrosourea-induced neurogenic tumors. I. Potentiation of tumor immunity withCorynebacterium parvum . J Neuropathol Exp Neurol. 1981;40:526. doi: 10.1097/00005072-198109000-00004. [DOI] [PubMed] [Google Scholar]
- 9.Denlinger R, Axler D, Koestner DA, Liss L. Tumor-specific transplantation immunity to intracerebral challenge with cells from a methylnitrosourea-induced brain tumor. J Med. 1975;6:249. [PubMed] [Google Scholar]
- 10.De Plaen E, Lurquin C, Van Pel A, Mariamé B, Szikora JP, Wölfel T, Sibille C, Chomez P, Boon T. Immunogenic (tum−) variants of mouse tumor P815: cloning of the gene of tum− antigen P91A and identification of the tum− mutation. Proc Natl Acad Sci USA. 1988;85:2274. doi: 10.1073/pnas.85.7.2274. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11.Frost P, Kerbel RS, Bauer E, Tartamello-Biondo R, Cefalu W. Mutagen treatment as a means for selecting immunogenic variants from otherwise poorly immunogenic malignant murine tumors. Cancer Res. 1983;43:125. [PubMed] [Google Scholar]
- 12.Grace JT, Perese DM, Metzgar RS, Sasabe T, Holdridge BS. Tumor autograft responses in patients with glioblastoma multiforme. J Neurosurg. 1961;18:159. [Google Scholar]
- 13.Greene HSN. Heterotransplantation of tumors. Ann NY Acad Sci. 1957;69:818. doi: 10.1111/j.1749-6632.1957.tb49720.x. [DOI] [PubMed] [Google Scholar]
- 14.Hewitt HB, Blake ER, Walder AS. A critique of the evidence for active host defense against cancer, based on personal studies of 27 murine tumours of spontaneous origin. Br J Cancer. 1976;33:241. doi: 10.1038/bjc.1976.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Itaya T, Hunt B, Frost P. Retention of immunogenicity after X-irradiation of mouse colon tumor cells expressing the transfected influenza virus hemagglutinin gene. Cancer Immunol Immunother. 1989;28:248. doi: 10.1007/BF00205233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16.Kaye A. Adjuvant treatment of malignant brain tumors. Aust NZ J Neurosurg. 1989;59:831. doi: 10.1111/j.1445-2197.1989.tb07023.x. [DOI] [PubMed] [Google Scholar]
- 17.Kida Y, Cravioto H, Hochwald GH, Hochgeschwender U, Ransohoff J. Immunity to transplantable nitrosourea-induced neurogenic tumors. II. Immunoprophylaxis of tumors of the brain. J Neuropathol Exp Neurol. 1983;42:122. doi: 10.1097/00005072-198303000-00002. [DOI] [PubMed] [Google Scholar]
- 18.Kelin G, Sjögren HO, Klein E, Hellström K. Demonstration of resistance against methylcholanthrene-induced sarcomas in the primary autochthonous host. Cancer Res. 1960;20:1561. [PubMed] [Google Scholar]
- 19.Koestner A, Swenberg JA, Wechsler W. Transplacental production with ethylnitrosourea of neoplasms of the nervous system in Sprague-Dawley rats. Am J Pathol. 1971;63:37. [PMC free article] [PubMed] [Google Scholar]
- 20.Lindvall M, Sjögren HO. Inhibition of yolk sac tumor growth in vivo by a monoclonal antibody to the retroviral molecule P15E. Cancer Immunol Immunother. 1991;33:21. doi: 10.1007/BF01742523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21.Long RG, O'Connor JS, Jelsma LF. Studies on glioma immunity in the mouse. Arch Neurol. 1962;7:76. [Google Scholar]
- 22.Lurquin C, Van Pel A, Mariamé B, De Plaen E, Szikora J, Janssens C, Reddehase MJ, Lejeune J, Boon T. Structure of the gene of tum− transplantation antigen P91A: the mutated exon encodes a peptide recognized with Ld by cytolytic T cells. Cell. 1989;58:293. doi: 10.1016/0092-8674(89)90844-1. [DOI] [PubMed] [Google Scholar]
- 23.Mahaley MS, Bigner DD, Dudka FL, Wilds PR, Williams DH, Bouldin TW, Whitaker JN, Bynum JM. Immunobiology of primary intracranial tumors. Part 7. Active immunization of patients with anaplastic human glioma cells: a pilot study. J Neurosurg. 1983;59:201. doi: 10.3171/jns.1983.59.2.0201. [DOI] [PubMed] [Google Scholar]
- 24.Maryanski J, Boon T. Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815 IV: analysis of variant-specific antigens by selection of antigen-loss variants with cytolytic T cell clones. Eur J Immunol. 1982;12:406. doi: 10.1002/eji.1830120509. [DOI] [PubMed] [Google Scholar]
- 25.Maryanski JL, Van Snick J, Cerottini JC, Boon T. Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815 III. Clonal analysis of the syngeneic T lymphocyte response. Eur J Immunol. 1982;12:401. doi: 10.1002/eji.1830120508. [DOI] [PubMed] [Google Scholar]
- 26.Medawar PB. Immunity to homologous grafted skin. III. The fate of skin homografts transplanted to the brain, to subcutaneous tissue and to the anterior chamber of the eye. Br J Exp Pathol. 1948;29:58. [PMC free article] [PubMed] [Google Scholar]
- 27.Merchant R, Ellison M, Young H. Immunotherapy for malignant glioma using human recombinant interleukin-2 and activated autologous lymphocytes. J Neuroonc. 1990;8:173. doi: 10.1007/BF00177842. [DOI] [PubMed] [Google Scholar]
- 28.Micco C, Hassoun J, Meyer G, Toga M. Role of the blood-brain barrier in the establishment of the immune response against polyoma virus-induced cerebral tumors in hamsters. J Neuroimmunol. 1986;11:301. doi: 10.1016/0165-5728(86)90083-4. [DOI] [PubMed] [Google Scholar]
- 29.Middle J, Robinson G, Embleton M. Naturally arising tumors of the inbred WAB/Not rat strain II. Immunogenicity of transplanted tumors. JNCI. 1981;67:637. [PubMed] [Google Scholar]
- 30.Nitta T, Sato K, Okumura K, Ishii S. Induction of cytotoxicity in human T-cells coated with anti-glioma x anti-CD3 bispecific antibody against human glioma cells. J Neurosurg. 1990;72:476. doi: 10.3171/jns.1990.72.3.0476. [DOI] [PubMed] [Google Scholar]
- 31.Ostrand-Rosenberg S, Thakur A, Clements V. Rejection of mouse sarcoma cells after transfection of MHC class II genes. J Immunol. 1990;144:4068. [PubMed] [Google Scholar]
- 32.Prehn R, Main J. Immunity to methylcholanthrene-induced sarcomas. JNCI. 1957;18:769. [PubMed] [Google Scholar]
- 33.Puccetti P, Bianchi R, Romani L, Cenci E, Fioretti MC. Delayed-type hypersensitivity to tumor antigens co-expressed with immunogenic determinants induced by xenogenization. Int J Cancer. 1989;43:279. doi: 10.1002/ijc.2910430220. [DOI] [PubMed] [Google Scholar]
- 34.Rainbird S, Ridley A. Antigenicity of ethylnitrosourea-induced rat schwannomas assayed by in vitro lymphocytotoxicity. Neuropathol Appl Neurobiol. 1977;3:9. [Google Scholar]
- 35.Salford L, Brun A, Nirfalk S. Ten-year survival among patients with supratentorial astrocytomas grade III and IV. J Neurosurg. 1988;69:506. doi: 10.3171/jns.1988.69.4.0506. [DOI] [PubMed] [Google Scholar]
- 36.Schackert HK, Itaya T, Schackert G, Fearon E, Vogelstein B, Frost F. Systemic immunity against a murine colon tumor (CT-26) produced by immunization with syngeneic cells expressing a transfected viral gene product. Int J Cancer. 1989;43:823. doi: 10.1002/ijc.2910430514. [DOI] [PubMed] [Google Scholar]
- 37.Scheinberg LC, Suzuki K, Edelman FL, Davidoff LM. Studies in immunization against a transplantable cerebral mouse glioma. J Neurosurg. 1963;4:312. doi: 10.3171/jns.1963.20.4.0312. [DOI] [PubMed] [Google Scholar]
- 38.Scheinberg LC, Edelman FL, Levy WA. Is the brain “an immunologically priveliged site”? I. Studies based on intracerebral tumor homotransplantation and isotransplantation to sensitized hosts. Arch Neurol. 1964;11:248. doi: 10.1001/archneur.1964.00460210026003. [DOI] [PubMed] [Google Scholar]
- 39.Scheinberg LC, Kotsilimbas DG, Karpf R, Mayer N. Is the brain “an immunologically privileged site?” III. Studies based on homologous skin grafts to the brain and the subcutaneous tissue. Arch Neurol. 1966;15:62. doi: 10.1001/archneur.1966.00470130066006. [DOI] [PubMed] [Google Scholar]
- 40.Sibille C, Chomez P, Wildmann C, Van Pel A, De Plaen E, Maryanski J, De Bergeryck V, Boon T. Structure of the gene of tum− transplantation antigen P198: a point mutation generates a new antigenic peptide. J Exp Med. 1990;172:35. doi: 10.1084/jem.172.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 41.Sjögren HO. Transplantation methods as a tool for detection of tumor-specific antigens. Prog Exp Tumor Res. 1965;6:289. [PubMed] [Google Scholar]
- 42.Spence AM, Priestly G. A survey of ethylnitrosourea-induced rat gliomas for the presence of tumor rejection antigens expressed in vivo. Neuropathol Appl Neurobiol. 1981;7:63. doi: 10.1111/j.1365-2990.1981.tb00232.x. [DOI] [PubMed] [Google Scholar]
- 43.Szikora JP, Van Pel A, Brichard V, Andre M, Van Baren N, Henry P, De Plaen E, Boon T. Structure of the gene of tum− transplantation antigen P35B: presence of a point mutation in the antigenic allele. EMBO J. 1990;9:1041. doi: 10.1002/j.1460-2075.1990.tb08208.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 44.Uyttenhove C, Van Snick J, Boon T. Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. I. Rejection by syngeneic mice. J Exp Med. 1980;152:1175. doi: 10.1084/jem.152.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 45.Van den Eynde B, Hainaut P, Hérin M, Knuth A, Lemoine C, Weynants P, Van der Bruggen P, Fauchet R, Boon T. Presence on a human melanoma of multiple antigens recognized by autologous lymphocytes. Int J Cancer. 1989;44:634. doi: 10.1002/ijc.2910440413. [DOI] [PubMed] [Google Scholar]
- 46.Van Pel A, Vessière F, Boon T. Protection against two spontaneous mouse leukemias conferred by immunogenic variants obtained by mutagenesis. J Exp Med. 1983;157:1992. doi: 10.1084/jem.157.6.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 47.Wekerle H, Engelhardt B, Risau W, Meyerman R. Pathophysiology of the blood-brain barrier. Amsterdam: Elsevier; 1990. Passage of lymphocytes across the blood-brain barrier in health and disease; p. 439. [Google Scholar]
- 48.Wen P, Lampson M, Lampson L. Effects of gamma-interferon on major histocompatibility complex antigen expression and lymphocytic infiltration in the 9L gliosarcoma brain tumor model: implications for strategies of immunotherapy. J Neuroimmunol. 1992;36:57. doi: 10.1016/0165-5728(92)90030-o. [DOI] [PubMed] [Google Scholar]
- 49.Yumitori K, Ito Y, Handa H. Protective effect of immunization with virus infected glioma cells against intracerebrally implanted glioma in mice. Eur J Clin Oncol. 1982;18:177. doi: 10.1016/0277-5379(82)90061-x. [DOI] [PubMed] [Google Scholar]