Assessment of major histocompatibility complex class I interaction with Epstein-Barr virus and human immunodeficiency virus peptides by elevation of membrane H-2 and HLA in peptide loading-deficient cells

Eur J Immunol. 1992 Oct;22(10):2697-703. doi: 10.1002/eji.1830221033.


Earlier findings indicate that peptides can affect the expression of major histocompatibility complex (MHC) class I molecules on the surface of cells with defective peptide loading mechanism. We have used peptide induced increase of class I antigen expression to assess peptide interaction with MHC class I molecules. A panel of 41 overlapping synthetic peptides derived from the human immunodeficiency virus-1 (HIV-1) gag protein and 33 nonoverlapping peptides from Epstein-Barr virus (EBV) proteins EBNA-1, 2, 3, 4, 5, 6, LMP, BZLF2, BILF2, BSLF2, BALF4 and BcLF1 was assessed for the ability to enhance the expression of HLA-A2.1, H-2Db, Kb and Dd on the murine RMA-S and human 721.174/T2 (.174/T2) lines by indirect immunofluorescence. Considering doubling of the fluorescence intensity in the peptide-treated samples as positivity, 6 of 39 HIV and 1 of 32 EBV peptides were found to bind to A2.1, 6 of 39 HIV gag and 7 of 16 EBV peptides to Db, 8 of 39 HIV gag and 5 of 16 EBV peptides to Kb and 2 of 39 HIV gag and 1 of 17 EBV peptides to Dd. The sensitivity of the method is comparable to the in vitro class I assembly assay with conformation-dependent monoclonal antibody and is more discriminating than the solid-phase assay. Due to its simplicity this method can also serve for testing large peptide panels for binding capacity to various class I molecules. Moreover, the method provides information about the relevance of in vitro tests for class I assembly in living cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, Viral / immunology*
  • DNA-Binding Proteins / immunology*
  • Epstein-Barr Virus Nuclear Antigens
  • Gene Products, gag / immunology*
  • H-2 Antigens / analysis*
  • HIV-1 / immunology*
  • HLA-A2 Antigen / analysis*
  • Herpesvirus 4, Human / immunology*
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Viral Proteins / immunology*


  • Antigens, Viral
  • DNA-Binding Proteins
  • Epstein-Barr Virus Nuclear Antigens
  • Gene Products, gag
  • H-2 Antigens
  • HLA-A2 Antigen
  • Viral Proteins