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. 1996 Aug 15;98(4):914–922. doi: 10.1172/JCI118874

A lung-specific neo-antigen elicits specific CD8+ T cell tolerance with preserved CD4+ T cell reactivity. Implications for immune-mediated lung disease.

R I Enelow 1, M H Stoler 1, A Srikiatkhachorn 1, C Kerlakian 1, S Agersborg 1, J A Whitsett 1, T J Braciale 1
PMCID: PMC507505  PMID: 8770862

Abstract

The A/Japan/57 influenza hemagglutinin (HA) was expressed in BALB/c mice under the transcriptional control of the surfactant protein C (SP-C) promoter, resulting in expression of HA in type II alveolar epithelial cells, as well as low level variable expression in other tissues, including the thymus in some of the founder lines. Transgenic animals were able to recover from infection with A/Japan/57 influenza, and they were able to mount antibody responses to A/Japan/57 HA in titers similar to wild type. We therefore tested their CD4+ T lymphocyte responses to HA and found them to be similar to wild type responses. However, CD8+ T cells from A/Japan/57-infected transgenic animals were unable to express cytolytic activity against target cells expressing the A/Japan/57 HA. The CD8+ T cell tolerance was also extremely specific, since transgenics immunized with an influenza strain containing a single amino acid substitution in a dominant HA epitope were able to mount full cytolytic responses to that epitope, but not the wild-type epitope. Adoptive transfer of CD8+ T cell clones into transgenic animals resulted extensive interstitial pneumonitis that was antigen-specific and associated with significant morbidity and mortality. We conclude that a lung-specific transgene may lead to specific CD8+ T cell tolerance, with CD4+ T cell and B cell reactivity to the antigen, and that CD4+ T cell reactivity may remain intact to an antigen expressed in the thymus, even when CD8+ T cell tolerance exists. This observation may have profound implications concerning immune-mediated lung diseases, particularly those mediated by CD4+ T cells.

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Selected References

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  1. Ada G. L., Jones P. D. The cell-mediated and humoral responses to influenza virus infection in the mouse lung. Adv Exp Med Biol. 1987;216B:1033–1042. [PubMed] [Google Scholar]
  2. Ada G. L., Jones P. D. The immune response to influenza infection. Curr Top Microbiol Immunol. 1986;128:1–54. doi: 10.1007/978-3-642-71272-2_1. [DOI] [PubMed] [Google Scholar]
  3. Anders E. M., Hartley C. A., Jackson D. C. Bovine and mouse serum beta inhibitors of influenza A viruses are mannose-binding lectins. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4485–4489. doi: 10.1073/pnas.87.12.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Askonas B. A. Our immune defence against influenza. Biochem Soc Trans. 1980 Jun;8(3):257–260. doi: 10.1042/bst0080257. [DOI] [PubMed] [Google Scholar]
  5. Berman J. S., Beer D. J., Theodore A. C., Kornfeld H., Bernardo J., Center D. M. Lymphocyte recruitment to the lung. Am Rev Respir Dis. 1990 Jul;142(1):238–257. doi: 10.1164/ajrccm/142.1.238. [DOI] [PubMed] [Google Scholar]
  6. Braciale T. J., Andrew M. E., Braciale V. L. Heterogeneity and specificity of cloned lines of influenza-virus specific cytotoxic T lymphocytes. J Exp Med. 1981 Apr 1;153(4):910–923. doi: 10.1084/jem.153.4.910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Braciale T. J., Braciale V. L. Viral antigen presentation and MHC assembly. Semin Immunol. 1992 Apr;4(2):81–84. [PubMed] [Google Scholar]
  8. Braciale T. J., Gerhard W., Klinman N. R. Analysis of the humoral immune response to influenza virus in vitro. J Immunol. 1976 Mar;116(3):827–834. [PubMed] [Google Scholar]
  9. Braciale T. J., Henkel T. J., Lukacher A., Braciale V. L. Fine specificity and antigen receptor expression among influenza virus-specific cytolytic T lymphocyte clones. J Immunol. 1986 Aug 1;137(3):995–1002. [PubMed] [Google Scholar]
  10. Braciale T. J. Immunologic recognition of influenza virus-infected cells. I. Generation of a virus-strain specific and a cross-reactive subpopulation of cytotoxic T cells in the response to type A influenza viruses of different subtypes. Cell Immunol. 1977 Oct;33(2):423–436. doi: 10.1016/0008-8749(77)90170-8. [DOI] [PubMed] [Google Scholar]
  11. Braciale T. J., Morrison L. A., Sweetser M. T., Sambrook J., Gething M. J., Braciale V. L. Antigen presentation pathways to class I and class II MHC-restricted T lymphocytes. Immunol Rev. 1987 Aug;98:95–114. doi: 10.1111/j.1600-065x.1987.tb00521.x. [DOI] [PubMed] [Google Scholar]
  12. Braciale T. J., Sweetser M. T., Brown L. R., Braciale V. L. T-lymphocyte recognition of a membrane glycoprotein. Cold Spring Harb Symp Quant Biol. 1989;54(Pt 1):477–485. doi: 10.1101/sqb.1989.054.01.057. [DOI] [PubMed] [Google Scholar]
  13. Braciale T. J., Sweetser M. T., Morrison L. A., Kittlesen D. J., Braciale V. L. Class I major histocompatibility complex-restricted cytolytic T lymphocytes recognize a limited number of sites on the influenza hemagglutinin. Proc Natl Acad Sci U S A. 1989 Jan;86(1):277–281. doi: 10.1073/pnas.86.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Brown L. R., Nygard N. R., Graham M. B., Bono C., Braciale V. L., Gorka J., Schwartz B. D., Braciale T. J. Recognition of the influenza hemagglutinin by class II MHC-restricted T lymphocytes and antibodies. I. Site definition and implications for antigen presentation and T lymphocyte recognition. J Immunol. 1991 Oct 15;147(8):2677–2684. [PubMed] [Google Scholar]
  15. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  16. Couch R. B., Kasel J. A. Immunity to influenza in man. Annu Rev Microbiol. 1983;37:529–549. doi: 10.1146/annurev.mi.37.100183.002525. [DOI] [PubMed] [Google Scholar]
  17. Cresswell P. Immunology: questions of presentation. Nature. 1990 Feb 15;343(6259):593–594. doi: 10.1038/343593a0. [DOI] [PubMed] [Google Scholar]
  18. Glasser S. W., Korfhagen T. R., Bruno M. D., Dey C., Whitsett J. A. Structure and expression of the pulmonary surfactant protein SP-C gene in the mouse. J Biol Chem. 1990 Dec 15;265(35):21986–21991. [PubMed] [Google Scholar]
  19. Gould K., Cossins J., Bastin J., Brownlee G. G., Townsend A. A 15 amino acid fragment of influenza nucleoprotein synthesized in the cytoplasm is presented to class I-restricted cytotoxic T lymphocytes. J Exp Med. 1989 Sep 1;170(3):1051–1056. doi: 10.1084/jem.170.3.1051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Graham M. B., Dalton D. K., Giltinan D., Braciale V. L., Stewart T. A., Braciale T. J. Response to influenza infection in mice with a targeted disruption in the interferon gamma gene. J Exp Med. 1993 Nov 1;178(5):1725–1732. doi: 10.1084/jem.178.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hämmerling G. J., Schönrich G., Ferber I., Arnold B. Peripheral tolerance as a multi-step mechanism. Immunol Rev. 1993 Jun;133:93–104. doi: 10.1111/j.1600-065x.1993.tb01511.x. [DOI] [PubMed] [Google Scholar]
  22. Kappler J. W., Roehm N., Marrack P. T cell tolerance by clonal elimination in the thymus. Cell. 1987 Apr 24;49(2):273–280. doi: 10.1016/0092-8674(87)90568-x. [DOI] [PubMed] [Google Scholar]
  23. Kisielow P., Blüthmann H., Staerz U. D., Steinmetz M., von Boehmer H. Tolerance in T-cell-receptor transgenic mice involves deletion of nonmature CD4+8+ thymocytes. Nature. 1988 Jun 23;333(6175):742–746. doi: 10.1038/333742a0. [DOI] [PubMed] [Google Scholar]
  24. Korfhagen T. R., Glasser S. W., Wert S. E., Bruno M. D., Daugherty C. C., McNeish J. D., Stock J. L., Potter S. S., Whitsett J. A. Cis-acting sequences from a human surfactant protein gene confer pulmonary-specific gene expression in transgenic mice. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6122–6126. doi: 10.1073/pnas.87.16.6122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ktistakis N. T., Thomas D., Roth M. G. Characteristics of the tyrosine recognition signal for internalization of transmembrane surface glycoproteins. J Cell Biol. 1990 Oct;111(4):1393–1407. doi: 10.1083/jcb.111.4.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lo D., Freedman J., Hesse S., Palmiter R. D., Brinster R. L., Sherman L. A. Peripheral tolerance to an islet cell-specific hemagglutinin transgene affects both CD4+ and CD8+ T cells. Eur J Immunol. 1992 Apr;22(4):1013–1022. doi: 10.1002/eji.1830220421. [DOI] [PubMed] [Google Scholar]
  27. Lukacher A. E., Braciale V. L., Braciale T. J. In vivo effector function of influenza virus-specific cytotoxic T lymphocyte clones is highly specific. J Exp Med. 1984 Sep 1;160(3):814–826. doi: 10.1084/jem.160.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lukacher A. E., Morrison L. A., Braciale V. L., Malissen B., Braciale T. J. Expression of specific cytolytic activity by H-2I region-restricted, influenza virus-specific T lymphocyte clones. J Exp Med. 1985 Jul 1;162(1):171–187. doi: 10.1084/jem.162.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. MacDonald H. R., Schneider R., Lees R. K., Howe R. C., Acha-Orbea H., Festenstein H., Zinkernagel R. M., Hengartner H. T-cell receptor V beta use predicts reactivity and tolerance to Mlsa-encoded antigens. Nature. 1988 Mar 3;332(6159):40–45. doi: 10.1038/332040a0. [DOI] [PubMed] [Google Scholar]
  30. Miller J. F., Heath W. R. Self-ignorance in the peripheral T-cell pool. Immunol Rev. 1993 Jun;133:131–150. doi: 10.1111/j.1600-065x.1993.tb01514.x. [DOI] [PubMed] [Google Scholar]
  31. Miller J. F., Morahan G., Allison J., Hoffmann M. A transgenic approach to the study of peripheral T-cell tolerance. Immunol Rev. 1991 Aug;122:103–116. doi: 10.1111/j.1600-065x.1991.tb00599.x. [DOI] [PubMed] [Google Scholar]
  32. Morrison L. A., Braciale V. L., Braciale T. J. Antigen form influences induction and frequency of influenza-specific class I and class II MHC-restricted cytolytic T lymphocytes. J Immunol. 1988 Jul 15;141(2):363–368. [PubMed] [Google Scholar]
  33. Morrison L. A., Braciale V. L., Braciale T. J. Distinguishable pathways of viral antigen presentation to T lymphocytes. Immunol Res. 1986;5(4):294–304. doi: 10.1007/BF02935502. [DOI] [PubMed] [Google Scholar]
  34. Morrison L. A., Lukacher A. E., Braciale V. L., Fan D. P., Braciale T. J. Differences in antigen presentation to MHC class I-and class II-restricted influenza virus-specific cytolytic T lymphocyte clones. J Exp Med. 1986 Apr 1;163(4):903–921. doi: 10.1084/jem.163.4.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Naim H. Y., Roth M. G. Characteristics of the internalization signal in the Y543 influenza virus hemagglutinin suggest a model for recognition of internalization signals containing tyrosine. J Biol Chem. 1994 Feb 11;269(6):3928–3933. [PubMed] [Google Scholar]
  36. Nossal G. J. Negative selection of lymphocytes. Cell. 1994 Jan 28;76(2):229–239. doi: 10.1016/0092-8674(94)90331-x. [DOI] [PubMed] [Google Scholar]
  37. Ohashi P. S., Oehen S., Buerki K., Pircher H., Ohashi C. T., Odermatt B., Malissen B., Zinkernagel R. M., Hengartner H. Ablation of "tolerance" and induction of diabetes by virus infection in viral antigen transgenic mice. Cell. 1991 Apr 19;65(2):305–317. doi: 10.1016/0092-8674(91)90164-t. [DOI] [PubMed] [Google Scholar]
  38. Oldstone M. B., Nerenberg M., Southern P., Price J., Lewicki H. Virus infection triggers insulin-dependent diabetes mellitus in a transgenic model: role of anti-self (virus) immune response. Cell. 1991 Apr 19;65(2):319–331. doi: 10.1016/0092-8674(91)90165-u. [DOI] [PubMed] [Google Scholar]
  39. Segal G. H., Shick H. E., Tubbs R. R., Fishleder A. J., Stoler M. H. In situ hybridization analysis of lymphoproliferative disorders. Assessment of clonality by immunoglobulin light-chain messenger RNA expression. Diagn Mol Pathol. 1994 Sep;3(3):170–177. [PubMed] [Google Scholar]
  40. Sweetser M. T., Braciale V. L., Braciale T. J. Class I major histocompatibility complex-restricted T lymphocyte recognition of the influenza hemagglutinin. Overlap between class I cytotoxic T lymphocytes and antibody sites. J Exp Med. 1989 Oct 1;170(4):1357–1368. doi: 10.1084/jem.170.4.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Walsh C. M., Matloubian M., Liu C. C., Ueda R., Kurahara C. G., Christensen J. L., Huang M. T., Young J. D., Ahmed R., Clark W. R. Immune function in mice lacking the perforin gene. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10854–10858. doi: 10.1073/pnas.91.23.10854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zinkernagel R. M., Althage A., Waterfield E., Kindred B., Welsh R. M., Callahan G., Pincetl P. Restriction specificities, alloreactivity, and allotolerance expressed by T cells from nude mice reconstituted with H-2-compatible or -incompatible thymus grafts. J Exp Med. 1980 Feb 1;151(2):376–399. doi: 10.1084/jem.151.2.376. [DOI] [PMC free article] [PubMed] [Google Scholar]

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