Abstract
One problem associated with the use of synthetic peptides as antigens in vivo is their susceptibility to inactivation by proteolytic degradation. A situation is described in which a serum protease, angiotensin-converting enzyme (ACE), is actually responsible for the class I binding activity of a commonly used influenza antigen, nucleoprotein (NP)(147-158R-). This peptide has been reported to be a highly efficient class I antigen. Evidence is presented that demonstrates that the peptide is inactive until cleaved by ACE, which is a normal constituent of serum. The enzyme removes a COOH-terminal dipeptide resulting in the sequence NP(147-155), which is identical to the naturally processed peptide. Such extracellular processing of peptides and proteins may occur for a variety of antigens both in vitro and in vivo, and could have important implications for the design of proteolytically resistant vaccines.
Full Text
The Full Text of this article is available as a PDF (536.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aichele P., Hengartner H., Zinkernagel R. M., Schulz M. Antiviral cytotoxic T cell response induced by in vivo priming with a free synthetic peptide. J Exp Med. 1990 May 1;171(5):1815–1820. doi: 10.1084/jem.171.5.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bodmer H. C., Pemberton R. M., Rothbard J. B., Askonas B. A. Enhanced recognition of a modified peptide antigen by cytotoxic T cells specific for influenza nucleoprotein. Cell. 1988 Jan 29;52(2):253–258. doi: 10.1016/0092-8674(88)90514-4. [DOI] [PubMed] [Google Scholar]
- Braciale T. J., Braciale V. L., Winkler M., Stroynowski I., Hood L., Sambrook J., Gething M. J. On the role of the transmembrane anchor sequence of influenza hemagglutinin in target cell recognition by class I MHC-restricted, hemagglutinin-specific cytolytic T lymphocytes. J Exp Med. 1987 Sep 1;166(3):678–692. doi: 10.1084/jem.166.3.678. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carbone F. R., Bevan M. J. Class I-restricted processing and presentation of exogenous cell-associated antigen in vivo. J Exp Med. 1990 Feb 1;171(2):377–387. doi: 10.1084/jem.171.2.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deres K., Schild H., Wiesmüller K. H., Jung G., Rammensee H. G. In vivo priming of virus-specific cytotoxic T lymphocytes with synthetic lipopeptide vaccine. Nature. 1989 Nov 30;342(6249):561–564. doi: 10.1038/342561a0. [DOI] [PubMed] [Google Scholar]
- Eisenlohr L. C., Yewdell J. W., Bennink J. R. Flanking sequences influence the presentation of an endogenously synthesized peptide to cytotoxic T lymphocytes. J Exp Med. 1992 Feb 1;175(2):481–487. doi: 10.1084/jem.175.2.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Falk K., Rötzschke O., Stevanović S., Jung G., Rammensee H. G. Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules. Nature. 1991 May 23;351(6324):290–296. doi: 10.1038/351290a0. [DOI] [PubMed] [Google Scholar]
- Friedland J., Setton C., Silverstein E. Induction of angiotensin converting enzyme in human monocytes in culture. Biochem Biophys Res Commun. 1978 Aug 14;83(3):843–849. doi: 10.1016/0006-291x(78)91471-7. [DOI] [PubMed] [Google Scholar]
- Gotch F., Rothbard J., Howland K., Townsend A., McMichael A. Cytotoxic T lymphocytes recognize a fragment of influenza virus matrix protein in association with HLA-A2. 1987 Apr 30-May 6Nature. 326(6116):881–882. doi: 10.1038/326881a0. [DOI] [PubMed] [Google Scholar]
- Hosken N. A., Bevan M. J. Defective presentation of endogenous antigen by a cell line expressing class I molecules. Science. 1990 Apr 20;248(4953):367–370. doi: 10.1126/science.2326647. [DOI] [PubMed] [Google Scholar]
- 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]
- Kane K. P., Sherman L. A., Mescher M. F. Exogenous beta 2-microglobulin is required for antigenic peptide binding to isolated class I major histocompatibility complex molecules. Eur J Immunol. 1991 Sep;21(9):2289–2292. doi: 10.1002/eji.1830210945. [DOI] [PubMed] [Google Scholar]
- Kast W. M., Roux L., Curren J., Blom H. J., Voordouw A. C., Meloen R. H., Kolakofsky D., Melief C. J. Protection against lethal Sendai virus infection by in vivo priming of virus-specific cytotoxic T lymphocytes with a free synthetic peptide. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2283–2287. doi: 10.1073/pnas.88.6.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krutzsch H. C. Determination of polypeptide amino acid sequences from the carboxyl terminus using angiotensin I converting enzyme. Biochemistry. 1980 Nov 11;19(23):5290–5296. doi: 10.1021/bi00564a022. [DOI] [PubMed] [Google Scholar]
- Maryanski J. L., Pala P., Corradin G., Jordan B. R., Cerottini J. C. H-2-restricted cytolytic T cells specific for HLA can recognize a synthetic HLA peptide. Nature. 1986 Dec 11;324(6097):578–579. doi: 10.1038/324578a0. [DOI] [PubMed] [Google Scholar]
- Moore M. W., Carbone F. R., Bevan M. J. Introduction of soluble protein into the class I pathway of antigen processing and presentation. Cell. 1988 Sep 9;54(6):777–785. doi: 10.1016/s0092-8674(88)91043-4. [DOI] [PubMed] [Google Scholar]
- Ondetti M. A., Cushman D. W. Enzymes of the renin-angiotensin system and their inhibitors. Annu Rev Biochem. 1982;51:283–308. doi: 10.1146/annurev.bi.51.070182.001435. [DOI] [PubMed] [Google Scholar]
- Parham P., Androlewicz M. J., Holmes N. J., Rothenberg B. E. Arginine 45 is a major part of the antigenic determinant of human beta 2-microglobulin recognized by mouse monoclonal antibody BBM.1. J Biol Chem. 1983 May 25;258(10):6179–6186. [PubMed] [Google Scholar]
- Rock K. L., Gamble S., Rothstein L., Benacerraf B. Reassociation with beta 2-microglobulin is necessary for Db class I major histocompatibility complex binding of an exogenous influenza peptide. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):301–304. doi: 10.1073/pnas.88.1.301. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rock K. L., Gamble S., Rothstein L. Presentation of exogenous antigen with class I major histocompatibility complex molecules. Science. 1990 Aug 24;249(4971):918–921. doi: 10.1126/science.2392683. [DOI] [PubMed] [Google Scholar]
- Rock K. L., Rothstein L. E., Gamble S. R., Benacerraf B. Reassociation with beta 2-microglobulin is necessary for Kb class I major histocompatibility complex binding of exogenous peptides. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7517–7521. doi: 10.1073/pnas.87.19.7517. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Staerz U. D., Karasuyama H., Garner A. M. Cytotoxic T lymphocytes against a soluble protein. Nature. 1987 Oct 1;329(6138):449–451. doi: 10.1038/329449a0. [DOI] [PubMed] [Google Scholar]
- Townsend A. R., Rothbard J., Gotch F. M., Bahadur G., Wraith D., McMichael A. J. The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell. 1986 Mar 28;44(6):959–968. doi: 10.1016/0092-8674(86)90019-x. [DOI] [PubMed] [Google Scholar]
- Townsend A., Bodmer H. Antigen recognition by class I-restricted T lymphocytes. Annu Rev Immunol. 1989;7:601–624. doi: 10.1146/annurev.iy.07.040189.003125. [DOI] [PubMed] [Google Scholar]
- 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]
- Vitiello A., Potter T. A., Sherman L. A. The role of beta 2-microglobulin in peptide binding by class I molecules. Science. 1990 Dec 7;250(4986):1423–1426. doi: 10.1126/science.2124002. [DOI] [PubMed] [Google Scholar]
- Vuk-Pavlović Z., Kreofsky T. J., Rohrbach M. S. Characteristics of monocyte angiotensin-converting enzyme (ACE) induction by dexamethasone. J Leukoc Biol. 1989 Jun;45(6):503–509. doi: 10.1002/jlb.45.6.503. [DOI] [PubMed] [Google Scholar]
- Widmann C., Maryanski J. L., Romero P., Corradin G. Differential stability of antigenic MHC class I-restricted synthetic peptides. J Immunol. 1991 Dec 1;147(11):3745–3751. [PubMed] [Google Scholar]
- Zijlstra M., Li E., Sajjadi F., Subramani S., Jaenisch R. Germ-line transmission of a disrupted beta 2-microglobulin gene produced by homologous recombination in embryonic stem cells. Nature. 1989 Nov 23;342(6248):435–438. doi: 10.1038/342435a0. [DOI] [PubMed] [Google Scholar]