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. 1996 Feb;70(2):1072–1079. doi: 10.1128/jvi.70.2.1072-1079.1996

CD4-deficient mice have reduced levels of memory cytotoxic T lymphocytes after immunization and show diminished resistance to subsequent virus challenge.

M G von Herrath 1, M Yokoyama 1, J Dockter 1, M B Oldstone 1, J L Whitton 1
PMCID: PMC189913  PMID: 8551565

Abstract

Although primary antiviral CD8+ cytotoxic T lymphocytes (CTL) can be induced in mice depleted of CD4+ T cells, the role of CD4+ T lymphocytes in the generation and maintenance of antiviral memory CTL is uncertain. This question, and the consequences upon vaccine-mediated protection, were investigated in transgenic CD4 knockout (CD4ko) mice, which lack CD4+ T lymphocytes. Infection of immunocompetent C57BL/6 mice with lymphocytic choriomeningitis virus (LCMV), or with recombinant vaccinia viruses bearing appropriate LCMV sequences, induces long-lasting protective immunity, mediated mainly by antiviral CD8+ CTL. Here we report two important findings. First, LCMV-specific CD8+ memory CTL are maintained at considerably lower levels in CD4ko mice than in normal C57BL/6J mice; we demonstrate a reduction in precursor CTL evident as soon as 30 days postimmunization and declining, by day 120, to levels 1 to 2 log units below those in normal mice. Thus, CD4+ T cells appear to be important to the generation and maintenance of their CD8+ counterparts. Second, this reduction has an important biological consequence; compared with immunocompetent mice, CD4ko mice immunized with vaccinia virus recombinants expressing nucleoprotein or glycoprotein of LCMV are less effectively protected from subsequent LCMV challenge. Thus, this study underscores the potential importance of CD4+ T lymphocytes in generation of appropriate levels of CD(8+)-cell-mediated immunoprotective memory and has implications for vaccine efficacy in individuals with immune defects in which CD4 levels may be reduced, such as AIDS.

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Selected References

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  1. Ahmed R., Butler L. D., Bhatti L. T4+ T helper cell function in vivo: differential requirement for induction of antiviral cytotoxic T-cell and antibody responses. J Virol. 1988 Jun;62(6):2102–2106. doi: 10.1128/jvi.62.6.2102-2106.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ahmed R., Byrne J. A., Oldstone M. B. Virus specificity of cytotoxic T lymphocytes generated during acute lymphocytic choriomeningitis virus infection: role of the H-2 region in determining cross-reactivity for different lymphocytic choriomeningitis virus strains. J Virol. 1984 Jul;51(1):34–41. doi: 10.1128/jvi.51.1.34-41.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baldridge J. R., Buchmeier M. J. Mechanisms of antibody-mediated protection against lymphocytic choriomeningitis virus infection: mother-to-baby transfer of humoral protection. J Virol. 1992 Jul;66(7):4252–4257. doi: 10.1128/jvi.66.7.4252-4257.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Battegay M., Moskophidis D., Rahemtulla A., Hengartner H., Mak T. W., Zinkernagel R. M. Enhanced establishment of a virus carrier state in adult CD4+ T-cell-deficient mice. J Virol. 1994 Jul;68(7):4700–4704. doi: 10.1128/jvi.68.7.4700-4704.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Borrow P., Evans C. F., Oldstone M. B. Virus-induced immunosuppression: immune system-mediated destruction of virus-infected dendritic cells results in generalized immune suppression. J Virol. 1995 Feb;69(2):1059–1070. doi: 10.1128/jvi.69.2.1059-1070.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Borrow P., Tishon A., Oldstone M. B. Infection of lymphocytes by a virus that aborts cytotoxic T lymphocyte activity and establishes persistent infection. J Exp Med. 1991 Jul 1;174(1):203–212. doi: 10.1084/jem.174.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doherty P. C., Hou S., Southern P. J. Lymphocytic choriomeningitis virus induces a chronic wasting disease in mice lacking class I major histocompatibility complex glycoproteins. J Neuroimmunol. 1993 Jul;46(1-2):11–17. doi: 10.1016/0165-5728(93)90228-q. [DOI] [PubMed] [Google Scholar]
  8. Eichelberger M., Allan W., Zijlstra M., Jaenisch R., Doherty P. C. Clearance of influenza virus respiratory infection in mice lacking class I major histocompatibility complex-restricted CD8+ T cells. J Exp Med. 1991 Oct 1;174(4):875–880. doi: 10.1084/jem.174.4.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GOOD R. A., ZAK S. J. Disturbances in gamma globulin synthesis as experiments of nature. Pediatrics. 1956 Jul;18(1):109–149. [PubMed] [Google Scholar]
  10. Gray D. Regulation of immunological memory. Curr Opin Immunol. 1994 Jun;6(3):425–430. doi: 10.1016/0952-7915(94)90122-8. [DOI] [PubMed] [Google Scholar]
  11. Gray M. M., Hann I. M., Glass S., Eden O. B., Jones P. M., Stevens R. F. Mortality and morbidity caused by measles in children with malignant disease attending four major treatment centres: a retrospective review. Br Med J (Clin Res Ed) 1987 Jul 4;295(6589):19–22. doi: 10.1136/bmj.295.6589.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horvat B., Loukides J. A., Anandan L., Brewer E., Flood P. M. Production of interleukin 2 and interleukin 4 by immune CD4-CD8+ and their role in the generation of antigen-specific cytotoxic T cells. Eur J Immunol. 1991 Aug;21(8):1863–1871. doi: 10.1002/eji.1830210813. [DOI] [PubMed] [Google Scholar]
  13. Hou S., Doherty P. C., Zijlstra M., Jaenisch R., Katz J. M. Delayed clearance of Sendai virus in mice lacking class I MHC-restricted CD8+ T cells. J Immunol. 1992 Aug 15;149(4):1319–1325. [PubMed] [Google Scholar]
  14. Hou S., Hyland L., Ryan K. W., Portner A., Doherty P. C. Virus-specific CD8+ T-cell memory determined by clonal burst size. Nature. 1994 Jun 23;369(6482):652–654. doi: 10.1038/369652a0. [DOI] [PubMed] [Google Scholar]
  15. Jennings S. R., Bonneau R. H., Smith P. M., Wolcott R. M., Chervenak R. CD4-positive T lymphocytes are required for the generation of the primary but not the secondary CD8-positive cytolytic T lymphocyte response to herpes simplex virus in C57BL/6 mice. Cell Immunol. 1991 Mar;133(1):234–252. doi: 10.1016/0008-8749(91)90194-g. [DOI] [PubMed] [Google Scholar]
  16. Jonjić S., del Val M., Keil G. M., Reddehase M. J., Koszinowski U. H. A nonstructural viral protein expressed by a recombinant vaccinia virus protects against lethal cytomegalovirus infection. J Virol. 1988 May;62(5):1653–1658. doi: 10.1128/jvi.62.5.1653-1658.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kasaian M. T., Leite-Morris K. A., Biron C. A. The role of CD4+ cells in sustaining lymphocyte proliferation during lymphocytic choriomeningitis virus infection. J Immunol. 1991 Mar 15;146(6):1955–1963. [PubMed] [Google Scholar]
  18. Kernahan J., McQuillin J., Craft A. W. Measles in children who have malignant disease. Br Med J (Clin Res Ed) 1987 Jul 4;295(6589):15–18. doi: 10.1136/bmj.295.6589.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Killeen N., Sawada S., Littman D. R. Regulated expression of human CD4 rescues helper T cell development in mice lacking expression of endogenous CD4. EMBO J. 1993 Apr;12(4):1547–1553. doi: 10.1002/j.1460-2075.1993.tb05798.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kirberg J., Bruno L., von Boehmer H. CD4+8- help prevents rapid deletion of CD8+ cells after a transient response to antigen. Eur J Immunol. 1993 Aug;23(8):1963–1967. doi: 10.1002/eji.1830230835. [DOI] [PubMed] [Google Scholar]
  21. Klavinskis L. S., Whitton J. L., Joly E., Oldstone M. B. Vaccination and protection from a lethal viral infection: identification, incorporation, and use of a cytotoxic T lymphocyte glycoprotein epitope. Virology. 1990 Oct;178(2):393–400. doi: 10.1016/0042-6822(90)90336-p. [DOI] [PubMed] [Google Scholar]
  22. Klavinskis L. S., Whitton J. L., Oldstone M. B. Molecularly engineered vaccine which expresses an immunodominant T-cell epitope induces cytotoxic T lymphocytes that confer protection from lethal virus infection. J Virol. 1989 Oct;63(10):4311–4316. doi: 10.1128/jvi.63.10.4311-4316.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kyburz D., Speiser D. E., Aebischer T., Hengartner H., Zinkernagel R. M. Virus-specific cytotoxic T cell-mediated lysis of lymphocytes in vitro and in vivo. J Immunol. 1993 Jun 1;150(11):5051–5058. [PubMed] [Google Scholar]
  24. Lau L. L., Jamieson B. D., Somasundaram T., Ahmed R. Cytotoxic T-cell memory without antigen. Nature. 1994 Jun 23;369(6482):648–652. doi: 10.1038/369648a0. [DOI] [PubMed] [Google Scholar]
  25. Lehmann-Grube F., Löhler J., Utermöhlen O., Gegin C. Antiviral immune responses of lymphocytic choriomeningitis virus-infected mice lacking CD8+ T lymphocytes because of disruption of the beta 2-microglobulin gene. J Virol. 1993 Jan;67(1):332–339. doi: 10.1128/jvi.67.1.332-339.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Leist T. P., Kohler M., Zinkernagel R. M. Impaired generation of anti-viral cytotoxicity against lymphocytic choriomeningitis and vaccinia virus in mice treated with CD4-specific monoclonal antibody. Scand J Immunol. 1989 Dec;30(6):679–686. doi: 10.1111/j.1365-3083.1989.tb02476.x. [DOI] [PubMed] [Google Scholar]
  27. Liu Y., Müllbacher A. The generation and activation of memory class I MHC restricted cytotoxic T cell responses to influenza A virus in vivo do not require CD4+ T cells. Immunol Cell Biol. 1989 Dec;67(Pt 6):413–420. doi: 10.1038/icb.1989.58. [DOI] [PubMed] [Google Scholar]
  28. MacDonald H. R., Cerottini J. C., Ryser J. E., Maryanski J. L., Taswell C., Widmer M. B., Brunner K. T. Quantitation and cloning of cytolytic T lymphocytes and their precursors. Immunol Rev. 1980;51:93–123. doi: 10.1111/j.1600-065x.1980.tb00318.x. [DOI] [PubMed] [Google Scholar]
  29. Matloubian M., Concepcion R. J., Ahmed R. CD4+ T cells are required to sustain CD8+ cytotoxic T-cell responses during chronic viral infection. J Virol. 1994 Dec;68(12):8056–8063. doi: 10.1128/jvi.68.12.8056-8063.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Matzinger P. Immunology. Memories are made of this? Nature. 1994 Jun 23;369(6482):605–606. doi: 10.1038/369605a0. [DOI] [PubMed] [Google Scholar]
  31. Mercadal C. M., Martin S., Rouse B. T. Apparent requirement for CD4+ T cells in primary anti-herpes simplex virus cytotoxic T-lymphocyte induction can be overcome by optimal antigen presentation. Viral Immunol. 1991 Fall;4(3):177–186. doi: 10.1089/vim.1991.4.177. [DOI] [PubMed] [Google Scholar]
  32. Mossman K., Upton C., Buller R. M., McFadden G. Species specificity of ectromelia virus and vaccinia virus interferon-gamma binding proteins. Virology. 1995 Apr 20;208(2):762–769. doi: 10.1006/viro.1995.1208. [DOI] [PubMed] [Google Scholar]
  33. Nahmias A. J., Griffith D., Salsbury C., Yoshida K. Thymic aplasia with lymphopenia, plasma cells, and normal immunoglobulins. Relation to measles virus infection. JAMA. 1967 Sep 4;201(10):729–734. [PubMed] [Google Scholar]
  34. Oldstone M. B., Blount P., Southern P. J., Lampert P. W. Cytoimmunotherapy for persistent virus infection reveals a unique clearance pattern from the central nervous system. Nature. 1986 May 15;321(6067):239–243. doi: 10.1038/321239a0. [DOI] [PubMed] [Google Scholar]
  35. Oldstone M. B., Salvato M., Tishon A., Lewicki H. Virus-lymphocyte interactions. III. Biologic parameters of a virus variant that fails to generate CTL and establishes persistent infection in immunocompetent hosts. Virology. 1988 Jun;164(2):507–516. doi: 10.1016/0042-6822(88)90565-x. [DOI] [PubMed] [Google Scholar]
  36. Oldstone M. B., Whitton J. L., Lewicki H., Tishon A. Fine dissection of a nine amino acid glycoprotein epitope, a major determinant recognized by lymphocytic choriomeningitis virus-specific class I-restricted H-2Db cytotoxic T lymphocytes. J Exp Med. 1988 Aug 1;168(2):559–570. doi: 10.1084/jem.168.2.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Palladino G., Scherle P. A., Gerhard W. Activity of CD4+ T-cell clones of type 1 and type 2 in generation of influenza virus-specific cytotoxic responses in vitro. J Virol. 1991 Nov;65(11):6071–6076. doi: 10.1128/jvi.65.11.6071-6076.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rahemtulla A., Fung-Leung W. P., Schilham M. W., Kündig T. M., Sambhara S. R., Narendran A., Arabian A., Wakeham A., Paige C. J., Zinkernagel R. M. Normal development and function of CD8+ cells but markedly decreased helper cell activity in mice lacking CD4. Nature. 1991 Sep 12;353(6340):180–184. doi: 10.1038/353180a0. [DOI] [PubMed] [Google Scholar]
  39. Rahemtulla A., Kündig T. M., Narendran A., Bachmann M. F., Julius M., Paige C. J., Ohashi P. S., Zinkernagel R. M., Mak T. W. Class II major histocompatibility complex-restricted T cell function in CD4-deficient mice. Eur J Immunol. 1994 Sep;24(9):2213–2218. doi: 10.1002/eji.1830240942. [DOI] [PubMed] [Google Scholar]
  40. Riviere Y., Oldstone M. B. Genetic reassortants of lymphocytic choriomeningitis virus: unexpected disease and mechanism of pathogenesis. J Virol. 1986 Aug;59(2):363–368. doi: 10.1128/jvi.59.2.363-368.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schulz M., Aichele P., Vollenweider M., Bobe F. W., Cardinaux F., Hengartner H., Zinkernagel R. M. Major histocompatibility complex--dependent T cell epitopes of lymphocytic choriomeningitis virus nucleoprotein and their protective capacity against viral disease. Eur J Immunol. 1989 Sep;19(9):1657–1667. doi: 10.1002/eji.1830190921. [DOI] [PubMed] [Google Scholar]
  42. Siegel M. M., Walter T. K., Ablin A. R. Measles pneumonia in childhood leukemia. Pediatrics. 1977 Jul;60(1):38–40. [PubMed] [Google Scholar]
  43. Smith G. L. Vaccinia virus glycoproteins and immune evasion. The sixteenth Fleming Lecture. J Gen Virol. 1993 Sep;74(Pt 9):1725–1740. doi: 10.1099/0022-1317-74-9-1725. [DOI] [PubMed] [Google Scholar]
  44. Sparshott S. M., Bell E. B. Membrane CD45R isoform exchange on CD4 T cells is rapid, frequent and dynamic in vivo. Eur J Immunol. 1994 Nov;24(11):2573–2578. doi: 10.1002/eji.1830241102. [DOI] [PubMed] [Google Scholar]
  45. Sprent J. Lifespans of naive, memory and effector lymphocytes. Curr Opin Immunol. 1993 Jun;5(3):433–438. doi: 10.1016/0952-7915(93)90065-z. [DOI] [PubMed] [Google Scholar]
  46. Spriggs M. K., Hruby D. E., Maliszewski C. R., Pickup D. J., Sims J. E., Buller R. M., VanSlyke J. Vaccinia and cowpox viruses encode a novel secreted interleukin-1-binding protein. Cell. 1992 Oct 2;71(1):145–152. doi: 10.1016/0092-8674(92)90273-f. [DOI] [PubMed] [Google Scholar]
  47. Tishon A., Lewicki H., Rall G., Von Herrath M., Oldstone M. B. An essential role for type 1 interferon-gamma in terminating persistent viral infection. Virology. 1995 Sep 10;212(1):244–250. doi: 10.1006/viro.1995.1477. [DOI] [PubMed] [Google Scholar]
  48. Ulmer J. B., Donnelly J. J., Parker S. E., Rhodes G. H., Felgner P. L., Dwarki V. J., Gromkowski S. H., Deck R. R., DeWitt C. M., Friedman A. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science. 1993 Mar 19;259(5102):1745–1749. doi: 10.1126/science.8456302. [DOI] [PubMed] [Google Scholar]
  49. Whitton J. L. Induction of protective immunity using minigenes. Ann N Y Acad Sci. 1994 Aug 15;730:107–117. doi: 10.1111/j.1749-6632.1994.tb44243.x. [DOI] [PubMed] [Google Scholar]
  50. Whitton J. L., Sheng N., Oldstone M. B., McKee T. A. A "string-of-beads" vaccine, comprising linked minigenes, confers protection from lethal-dose virus challenge. J Virol. 1993 Jan;67(1):348–352. doi: 10.1128/jvi.67.1.348-352.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Whitton J. L., Southern P. J., Oldstone M. B. Analyses of the cytotoxic T lymphocyte responses to glycoprotein and nucleoprotein components of lymphocytic choriomeningitis virus. Virology. 1988 Feb;162(2):321–327. doi: 10.1016/0042-6822(88)90471-0. [DOI] [PubMed] [Google Scholar]
  52. Whitton J. L., Tishon A., Lewicki H., Gebhard J., Cook T., Salvato M., Joly E., Oldstone M. B. Molecular analyses of a five-amino-acid cytotoxic T-lymphocyte (CTL) epitope: an immunodominant region which induces nonreciprocal CTL cross-reactivity. J Virol. 1989 Oct;63(10):4303–4310. doi: 10.1128/jvi.63.10.4303-4310.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wright K. E., Buchmeier M. J. Antiviral antibodies attenuate T-cell-mediated immunopathology following acute lymphocytic choriomeningitis virus infection. J Virol. 1991 Jun;65(6):3001–3006. doi: 10.1128/jvi.65.6.3001-3006.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. von Herrath M. G., Dockter J., Oldstone M. B. How virus induces a rapid or slow onset insulin-dependent diabetes mellitus in a transgenic model. Immunity. 1994 Jun;1(3):231–242. doi: 10.1016/1074-7613(94)90101-5. [DOI] [PubMed] [Google Scholar]

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