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J Exp Med. 1995 Dec 1; 182(6): 1751–1757.
PMCID: PMC2192246

Identification of a CD4+ T cell-stimulating antigen of pathogenic bacteria by expression cloning


Identifying the immunogenic proteins that elicit pathogen-specific T cell responses is key to rational vaccine design. While several approaches have succeeded in identifying major histocompatibility complex (MHC) class I bound peptides that stimulate CD8+ T cells, these approaches have been difficult to extend to peptides presented by MHC class II molecules that stimulate CD4+ T cells. We describe here a novel strategy for identifying CD4+ T cell-stimulating antigen genes. Using Listeria monocytogenes-specific, lacZ-inducible T cells as single- cell probes, we screened a Listeria monocytogenes genomic library as recombinant Escherichia coli that were fed to macrophages. The antigen gene was isolated from the E. coli clone that, when ingested by the macrophages, allowed generation of the appropriate peptide/MHC class II complex and T cell activation. We show that the antigenic peptide is derived from a previously unknown listeria gene product with characteristics of a membrane-bound protein.

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Selected References

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  • MACKANESS GB. Cellular resistance to infection. J Exp Med. 1962 Sep 1;116:381–406. [PMC free article] [PubMed]
  • Kaufmann SH. Immunity to intracellular bacteria. Annu Rev Immunol. 1993;11:129–163. [PubMed]
  • Sinha AA, Lopez MT, McDevitt HO. Autoimmune diseases: the failure of self tolerance. Science. 1990 Jun 15;248(4961):1380–1388. [PubMed]
  • Rudensky AYu, Preston-Hurlburt P, Hong SC, Barlow A, Janeway CA., Jr Sequence analysis of peptides bound to MHC class II molecules. Nature. 1991 Oct 17;353(6345):622–627. [PubMed]
  • Chicz RM, Urban RG, Lane WS, Gorga JC, Stern LJ, Vignali DA, Strominger JL. 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. [PubMed]
  • Hunt DF, Michel H, Dickinson TA, Shabanowitz J, Cox AL, Sakaguchi K, Appella E, Grey HM, 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. [PubMed]
  • van der Bruggen P, Traversari C, Chomez P, Lurquin C, De Plaen E, Van den Eynde B, Knuth A, Boon T. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science. 1991 Dec 13;254(5038):1643–1647. [PubMed]
  • Pamer EG, Harty JT, Bevan MJ. Precise prediction of a dominant class I MHC-restricted epitope of Listeria monocytogenes. Nature. 1991 Oct 31;353(6347):852–855. [PMC free article] [PubMed]
  • Udaka K, Tsomides TJ, Eisen HN. A naturally occurring peptide recognized by alloreactive CD8+ cytotoxic T lymphocytes in association with a class I MHC protein. Cell. 1992 Jun 12;69(6):989–998. [PubMed]
  • Henderson RA, Cox AL, Sakaguchi K, Appella E, Shabanowitz J, Hunt DF, Engelhard VH. Direct identification of an endogenous peptide recognized by multiple HLA-A2.1-specific cytotoxic T cells. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10275–10279. [PMC free article] [PubMed]
  • Malarkannan S, Afkarian M, Shastri N. A rare cryptic translation product is presented by Kb major histocompatibility complex class I molecule to alloreactive T cells. J Exp Med. 1995 Dec 1;182(6):1739–1750. [PMC free article] [PubMed]
  • Monach PA, Meredith SC, Siegel CT, Schreiber H. A unique tumor antigen produced by a single amino acid substitution. Immunity. 1995 Jan;2(1):45–59. [PubMed]
  • Nelson CA, Roof RW, McCourt DW, Unanue ER. Identification of the naturally processed form of hen egg white lysozyme bound to the murine major histocompatibility complex class II molecule I-Ak. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7380–7383. [PMC free article] [PubMed]
  • Karttunen J, Shastri N. Measurement of ligand-induced activation in single viable T cells using the lacZ reporter gene. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3972–3976. [PMC free article] [PubMed]
  • Karttunen J, Sanderson S, Shastri N. Detection of rare antigen-presenting cells by the lacZ T-cell activation assay suggests an expression cloning strategy for T-cell antigens. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6020–6024. [PMC free article] [PubMed]
  • Sanderson S, Shastri N. LacZ inducible, antigen/MHC-specific T cell hybrids. Int Immunol. 1994 Mar;6(3):369–376. [PubMed]
  • Brunt LM, Portnoy DA, Unanue ER. Presentation of Listeria monocytogenes to CD8+ T cells requires secretion of hemolysin and intracellular bacterial growth. J Immunol. 1990 Dec 1;145(11):3540–3546. [PubMed]
  • De Bruijn ML, Nieland JD, Harding CV, Melief CJ. Processing and presentation of intact hen egg-white lysozyme by dendritic cells. Eur J Immunol. 1992 Sep;22(9):2347–2352. [PubMed]
  • Pfeifer JD, Wick MJ, Russell DG, Normark SJ, Harding CV. Recombinant Escherichia coli express a defined, cytoplasmic epitope that is efficiently processed in macrophage phagolysosomes for class II MHC presentation to T lymphocytes. J Immunol. 1992 Oct 15;149(8):2576–2584. [PubMed]
  • Rasmussen OF, Beck T, Olsen JE, Dons L, Rossen L. Listeria monocytogenes isolates can be classified into two major types according to the sequence of the listeriolysin gene. Infect Immun. 1991 Nov;59(11):3945–3951. [PMC free article] [PubMed]
  • Bancroft GJ, Schreiber RD, Unanue ER. Natural immunity: a T-cell-independent pathway of macrophage activation, defined in the scid mouse. Immunol Rev. 1991 Dec;124:5–24. [PubMed]
  • Kaufmann SH, Hug E, Väth U, Müller I. Effective protection against Listeria monocytogenes and delayed-type hypersensitivity to listerial antigens depend on cooperation between specific L3T4+ and Lyt 2+ T cells. Infect Immun. 1985 Apr;48(1):263–266. [PMC free article] [PubMed]
  • Kaufmann SH, Simon MM, Hahn H. Specific Lyt 123 cells are involved in protection against Listeria monocytogenes and in delayed-type hypersensitivity to listerial antigens. J Exp Med. 1979 Oct 1;150(4):1033–1038. [PMC free article] [PubMed]
  • Ladel CH, Flesch IE, Arnoldi J, Kaufmann SH. Studies with MHC-deficient knock-out mice reveal impact of both MHC I- and MHC II-dependent T cell responses on Listeria monocytogenes infection. J Immunol. 1994 Oct 1;153(7):3116–3122. [PubMed]
  • Pamer EG. Direct sequence identification and kinetic analysis of an MHC class I-restricted Listeria monocytogenes CTL epitope. J Immunol. 1994 Jan 15;152(2):686–694. [PubMed]
  • Beattie IA, Swaminathan B, Ziegler HK. Cloning and characterization of T-cell-reactive protein antigens from Listeria monocytogenes. Infect Immun. 1990 Sep;58(9):2792–2803. [PMC free article] [PubMed]
  • von Heijne G. The structure of signal peptides from bacterial lipoproteins. Protein Eng. 1989 May;2(7):531–534. [PubMed]
  • Hayashi S, Wu HC. Lipoproteins in bacteria. J Bioenerg Biomembr. 1990 Jun;22(3):451–471. [PubMed]
  • Bouwer HG, Gibbins BL, Jones S, Hinrichs DJ. Antilisterial immunity includes specificity to listeriolysin O (LLO) and non-LLO-derived determinants. Infect Immun. 1994 Mar;62(3):1039–1045. [PMC free article] [PubMed]
  • Daugelat S, Ladel CH, Schoel B, Kaufmann SH. Antigen-specific T-cell responses during primary and secondary Listeria monocytogenes infection. Infect Immun. 1994 May;62(5):1881–1888. [PMC free article] [PubMed]

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