Quantitative and qualitative influences of tapasin on the class I peptide repertoire

A. W. Purcell, J. J. Gorman, M. Garcia-Peydró, A. Paradela, S. R. Burrows, G. H. Talbo, N. Laham, Au Peh Chen Au Peh, E. C. Reynolds, J. A. López de Castro, J. McCluskey

Research output: Contribution to journalArticleResearchpeer-review

127 Citations (Scopus)

Abstract

Tapasin is critical for efficient loading and surface expression of most HLA class I molecules. The high level surface expression of HLA-B*2705 on tapasin-deficient 721.220 cells allowed the influence of this chaperone on peptide repertoire to be examined. Comparison of peptides bound to HLA-B*2705 expressed on tapasin-deficient and -proficient cells by mass spectrometry revealed an overall reduction in the recovery of B*2705-bound peptides isolated from tapasin-deficient cells despite similar yields of B27 heavy chain and β2-microglobulin. This indicated that a proportion of suboptimal ligands were associated with B27, and they were lost during the purification process. Notwithstanding this failure to recover these suboptimal peptides, there was substantial overlap in the repertoire and biochemical properties of peptides recovered from B27 complexes derived from tapasin-positive and -negative cells. Although many peptides were preferentially or uniquely isolated from B*2705 in tapasin-positive cells, a number of species were preferentially recovered in the absence of tapasin, and some of these peptide ligands have been sequenced. In general, these ligands did not exhibit exceptional binding affinity, and we invoke an argument based on lumenal availability and affinity to explain their tapasin independence. The differential display of peptides in tapasin-negative and -positive cells was also in the reactivity of peptide-sensitive alloreactive CTL raised against tapasin-positive and -negative targets, demonstrating the functional relevance of the biochemical observation of changes in peptide repertoire in the tapasin-deficient APC. Overall, the data reveal that tapasin quantitatively and qualitatively influences ligand selection by class I molecules.

Original languageEnglish
Pages (from-to)1016-1027
Number of pages12
JournalJournal of Immunology
Volume166
Issue number2
DOIs
Publication statusPublished - 15 Jan 2001
Externally publishedYes

Cite this

Purcell, A. W., Gorman, J. J., Garcia-Peydró, M., Paradela, A., Burrows, S. R., Talbo, G. H., Laham, N., Chen Au Peh, A. P., Reynolds, E. C., López de Castro, J. A., & McCluskey, J. (2001). Quantitative and qualitative influences of tapasin on the class I peptide repertoire. Journal of Immunology, 166(2), 1016-1027. https://doi.org/10.4049/jimmunol.166.2.1016