Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27
<p><strong>Objective.</strong> HLA-B27 and ERAP1 are the two strongest predisposing genetic factors to Ankylosing Spondylitis (AS). A key aminopeptidase in MHC class I presentation, ERAP1 potentially contributes to AS pathogenesis through altering HLA-B27 peptide presentation. We s...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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Wiley-Blackwell
2013
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| _version_ | 1797095051179851776 |
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| author | Chen, L Fischer, R Peng, Y Reeves, E McHugh, K Ternette, N Hanke, T Dong, T Elliott, T Shastri, N Kollnberger, S James, E Bowness, P |
| author2 | American College of Rheumatology |
| author_facet | American College of Rheumatology Chen, L Fischer, R Peng, Y Reeves, E McHugh, K Ternette, N Hanke, T Dong, T Elliott, T Shastri, N Kollnberger, S James, E Bowness, P |
| author_sort | Chen, L |
| collection | OXFORD |
| description | <p><strong>Objective.</strong> HLA-B27 and ERAP1 are the two strongest predisposing genetic factors to Ankylosing Spondylitis (AS). A key aminopeptidase in MHC class I presentation, ERAP1 potentially contributes to AS pathogenesis through altering HLA-B27 peptide presentation. We studied the effects of ERAP1 on the HLA-B27 peptide repertoire and peptide presentation to Cytotoxic T lymphocytes (CTLs).</p> <p><strong>Methods.</strong> ERAP1-silenced and -competent HeLa.B27/C1R.B27 cells were isotope labeled, mixed, lysed and then immuno-precipitated using W6/32 or ME1. Peptides bound to HLA-B27 were eluted and analyzed by tandem Mass Spectrometry. Selected peptides were synthesized and tested for HLA-B27 binding ability. The effect of ERAP1 silencing/mutation on presentation of an immunodominant viral HLA-B27 epitope, KK10, to CTL was also studied.</p> <p><strong>Results.</strong> In both HeLa.B27 and C1R.B27 cells, the proportion of 9mer HLA-B27-bound peptides was decreased by ERAP1 silencing, whereas the percentage of longer peptides (11-13mers) increased. Surprisingly, following ERAP1 silencing, C-terminally extended peptides were readily identified. These were better able to bind to HLA-B27 than N-terminally extended peptides lacking a P2 Arginine. In both HeLa.B27 and mouse fibroblasts expressing HLA-B27, the absence of ERAP1 reduced recognition by HLA-B27-restricted KK10-specific CTLs following recombinant vaccinia viral infection or transfection with minigenes expressing KK10 precursors. Lastly, an AS protective variant, K528R-ERAP1, reduced KK10 CTL recognition following extended-KK10 minigene transfection compared to WT-ERAP1.</p> <p><strong>Conclusion.</strong> Our study shows that ERAP1 directly alters peptide binding and presentation by HLA-B27, supporting a pathogenic mechanism in AS. ERAP1 inhibition could potentially be used for treatment of AS and other ERAP1-associated diseases. © 2013 American College of Rheumatology.</p> |
| first_indexed | 2024-03-07T04:22:30Z |
| format | Journal article |
| id | oxford-uuid:cb7e8621-7cb2-4c3a-b4bf-ac89ca9d7d6e |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T04:22:30Z |
| publishDate | 2013 |
| publisher | Wiley-Blackwell |
| record_format | dspace |
| spelling | oxford-uuid:cb7e8621-7cb2-4c3a-b4bf-ac89ca9d7d6e2022-03-27T07:15:18ZEndoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:cb7e8621-7cb2-4c3a-b4bf-ac89ca9d7d6eMedical sciencesImmunologyRheumotologyEnglishOxford University Research Archive - ValetWiley-Blackwell2013Chen, LFischer, RPeng, YReeves, EMcHugh, KTernette, NHanke, TDong, TElliott, TShastri, NKollnberger, SJames, EBowness, PAmerican College of Rheumatology<p><strong>Objective.</strong> HLA-B27 and ERAP1 are the two strongest predisposing genetic factors to Ankylosing Spondylitis (AS). A key aminopeptidase in MHC class I presentation, ERAP1 potentially contributes to AS pathogenesis through altering HLA-B27 peptide presentation. We studied the effects of ERAP1 on the HLA-B27 peptide repertoire and peptide presentation to Cytotoxic T lymphocytes (CTLs).</p> <p><strong>Methods.</strong> ERAP1-silenced and -competent HeLa.B27/C1R.B27 cells were isotope labeled, mixed, lysed and then immuno-precipitated using W6/32 or ME1. Peptides bound to HLA-B27 were eluted and analyzed by tandem Mass Spectrometry. Selected peptides were synthesized and tested for HLA-B27 binding ability. The effect of ERAP1 silencing/mutation on presentation of an immunodominant viral HLA-B27 epitope, KK10, to CTL was also studied.</p> <p><strong>Results.</strong> In both HeLa.B27 and C1R.B27 cells, the proportion of 9mer HLA-B27-bound peptides was decreased by ERAP1 silencing, whereas the percentage of longer peptides (11-13mers) increased. Surprisingly, following ERAP1 silencing, C-terminally extended peptides were readily identified. These were better able to bind to HLA-B27 than N-terminally extended peptides lacking a P2 Arginine. In both HeLa.B27 and mouse fibroblasts expressing HLA-B27, the absence of ERAP1 reduced recognition by HLA-B27-restricted KK10-specific CTLs following recombinant vaccinia viral infection or transfection with minigenes expressing KK10 precursors. Lastly, an AS protective variant, K528R-ERAP1, reduced KK10 CTL recognition following extended-KK10 minigene transfection compared to WT-ERAP1.</p> <p><strong>Conclusion.</strong> Our study shows that ERAP1 directly alters peptide binding and presentation by HLA-B27, supporting a pathogenic mechanism in AS. ERAP1 inhibition could potentially be used for treatment of AS and other ERAP1-associated diseases. © 2013 American College of Rheumatology.</p> |
| spellingShingle | Medical sciences Immunology Rheumotology Chen, L Fischer, R Peng, Y Reeves, E McHugh, K Ternette, N Hanke, T Dong, T Elliott, T Shastri, N Kollnberger, S James, E Bowness, P Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27 |
| title | Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27 |
| title_full | Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27 |
| title_fullStr | Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27 |
| title_full_unstemmed | Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27 |
| title_short | Endoplasmic reticulum aminopeptidase 1 (ERAP1) plays a critical role in determining the length and sequence of peptides bound and presented by HLA-B27 |
| title_sort | endoplasmic reticulum aminopeptidase 1 erap1 plays a critical role in determining the length and sequence of peptides bound and presented by hla b27 |
| topic | Medical sciences Immunology Rheumotology |
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