MHC-peptide-specific antibodies reveal inefficient presentation of an HLA-A*0201-restricted, Melan-A-derived peptide after active intracellular processing
MHC-peptide-specific Fab antibodies binding to HLA-A*0201 complexes presenting the wild-type EAAGIGILTV (EAA) or analogue Melan-A 10-mer ELAGIGILTV (ELA) peptide were generated to study efficacy of peptide processing and presentation. None of the selected Fab antibodies detected the naturally processed EAA/HLA-A*0201 complex on melanoma tumor cells, confirming the known low peptide number on the cell surface. To study the effect of peptide presentation and processing in more detail, genes coding for the A27L-mutated Melan-A protein or the processed ELA peptide were introduced into HLA-A*0201(+) B cells by infection with the respective recombinant vaccinia virus construct producing equimolar amounts of GFP-ubiquitin directly linked to the fragment of interest. Correlating GFP expression to actual numbers of peptide presented, 1100-2600 [corrected] ELA peptides had to be synthesized to be presented by a single MHC class I antigen-peptide complex. This number increased 10- to 20-fold when ELA peptide presentation from the A27L-mutated full length Melan-A protein was studied, since 16000-52000 [corrected] GFP molecules needed to be synthesized for the detection of one ELA peptide. Our results indicate that peptide processing rather than presentation is the rate-limiting step in our experimental setting and is much more ineffective for Melan-A than has been previously shown for other MHC class I-restricted epitopes.
Errataetall: | |
---|---|
Medienart: |
Artikel |
Erscheinungsjahr: |
2007 |
---|---|
Erschienen: |
2007 |
Enthalten in: |
Zur Gesamtaufnahme - volume:37 |
---|---|
Enthalten in: |
European journal of immunology - 37(2007), 7 vom: 01. Juli, Seite 2008-17 |
Sprache: |
Englisch |
---|
Beteiligte Personen: |
Held, Gerhard [VerfasserIn] |
---|
Anmerkungen: |
Date Completed 14.09.2007 Date Revised 29.01.2022 published: Print ErratumIn: Eur J Immunol. 2008 May;38(5):1465-6 Citation Status MEDLINE |
---|
Förderinstitution / Projekttitel: |
|
---|
PPN (Katalog-ID): |
NLM170794741 |
---|
LEADER | 01000naa a22002652 4500 | ||
---|---|---|---|
001 | NLM170794741 | ||
003 | DE-627 | ||
005 | 20231223124304.0 | ||
007 | tu | ||
008 | 231223s2007 xx ||||| 00| ||eng c | ||
028 | 5 | 2 | |a pubmed24n0569.xml |
035 | |a (DE-627)NLM170794741 | ||
035 | |a (NLM)17559180 | ||
040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
041 | |a eng | ||
100 | 1 | |a Held, Gerhard |e verfasserin |4 aut | |
245 | 1 | 0 | |a MHC-peptide-specific antibodies reveal inefficient presentation of an HLA-A*0201-restricted, Melan-A-derived peptide after active intracellular processing |
264 | 1 | |c 2007 | |
336 | |a Text |b txt |2 rdacontent | ||
337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
338 | |a Band |b nc |2 rdacarrier | ||
500 | |a Date Completed 14.09.2007 | ||
500 | |a Date Revised 29.01.2022 | ||
500 | |a published: Print | ||
500 | |a ErratumIn: Eur J Immunol. 2008 May;38(5):1465-6 | ||
500 | |a Citation Status MEDLINE | ||
520 | |a MHC-peptide-specific Fab antibodies binding to HLA-A*0201 complexes presenting the wild-type EAAGIGILTV (EAA) or analogue Melan-A 10-mer ELAGIGILTV (ELA) peptide were generated to study efficacy of peptide processing and presentation. None of the selected Fab antibodies detected the naturally processed EAA/HLA-A*0201 complex on melanoma tumor cells, confirming the known low peptide number on the cell surface. To study the effect of peptide presentation and processing in more detail, genes coding for the A27L-mutated Melan-A protein or the processed ELA peptide were introduced into HLA-A*0201(+) B cells by infection with the respective recombinant vaccinia virus construct producing equimolar amounts of GFP-ubiquitin directly linked to the fragment of interest. Correlating GFP expression to actual numbers of peptide presented, 1100-2600 [corrected] ELA peptides had to be synthesized to be presented by a single MHC class I antigen-peptide complex. This number increased 10- to 20-fold when ELA peptide presentation from the A27L-mutated full length Melan-A protein was studied, since 16000-52000 [corrected] GFP molecules needed to be synthesized for the detection of one ELA peptide. Our results indicate that peptide processing rather than presentation is the rate-limiting step in our experimental setting and is much more ineffective for Melan-A than has been previously shown for other MHC class I-restricted epitopes | ||
650 | 4 | |a Journal Article | |
650 | 4 | |a Research Support, Non-U.S. Gov't | |
650 | 7 | |a Antigens, Neoplasm |2 NLM | |
650 | 7 | |a HLA-A Antigens |2 NLM | |
650 | 7 | |a HLA-A*02:01 antigen |2 NLM | |
650 | 7 | |a HLA-A2 Antigen |2 NLM | |
650 | 7 | |a Immunoglobulin Fab Fragments |2 NLM | |
650 | 7 | |a MART-1 Antigen |2 NLM | |
650 | 7 | |a MLANA protein, human |2 NLM | |
650 | 7 | |a Neoplasm Proteins |2 NLM | |
650 | 7 | |a Peptides |2 NLM | |
700 | 1 | |a Wadle, Andreas |e verfasserin |4 aut | |
700 | 1 | |a Dauth, Nina |e verfasserin |4 aut | |
700 | 1 | |a Stewart-Jones, Guillaume |e verfasserin |4 aut | |
700 | 1 | |a Sturm, Christine |e verfasserin |4 aut | |
700 | 1 | |a Thiel, Markus |e verfasserin |4 aut | |
700 | 1 | |a Zwick, Carsten |e verfasserin |4 aut | |
700 | 1 | |a Dieckmann, Detlef |e verfasserin |4 aut | |
700 | 1 | |a Schuler, Gerold |e verfasserin |4 aut | |
700 | 1 | |a Hoogenboom, Hennie R |e verfasserin |4 aut | |
700 | 1 | |a Lévy, Frédéric |e verfasserin |4 aut | |
700 | 1 | |a Cerundolo, Vincenzo |e verfasserin |4 aut | |
700 | 1 | |a Pfreundschuh, Michael |e verfasserin |4 aut | |
700 | 1 | |a Renner, Christoph |e verfasserin |4 aut | |
773 | 0 | 8 | |i Enthalten in |t European journal of immunology |d 1971 |g 37(2007), 7 vom: 01. Juli, Seite 2008-17 |w (DE-627)NLM000108723 |x 1521-4141 |7 nnns |
773 | 1 | 8 | |g volume:37 |g year:2007 |g number:7 |g day:01 |g month:07 |g pages:2008-17 |
912 | |a GBV_USEFLAG_A | ||
912 | |a GBV_NLM | ||
951 | |a AR | ||
952 | |d 37 |j 2007 |e 7 |b 01 |c 07 |h 2008-17 |