Hemophilia A subjects with an intron-22 gene inversion mutation show CD4
epitope mapping
factor VIII
hemophilia A
immune tolerance
intron-22 inversion mutation
Journal
Frontiers in immunology
ISSN: 1664-3224
Titre abrégé: Front Immunol
Pays: Switzerland
ID NLM: 101560960
Informations de publication
Date de publication:
2023
2023
Historique:
received:
21
12
2022
accepted:
20
02
2023
entrez:
20
3
2023
pubmed:
21
3
2023
medline:
22
3
2023
Statut:
epublish
Résumé
Almost half of severe hemophilia A (HA) is caused by an intron 22 inversion mutation (Int22Inv), which disrupts the 26-exon To test the hypothesis that (putative) intracellular synthesis of FVIII proteins encoded by inverted Peripheral blood mononuclear cells (PBMCs) from 30 severe or moderate HA subjects (17 with an Int22Inv mutation) were tested by ELISPOT assays to detect cytokine secretion in response to FVIII proteins and peptides and to map immunodominant T-cell epitopes. Potential immunogenicity of FVIII sequences encoded by the Eight of the Int22Inv subjects showed robust cytokine secretion from PBMCs stimulated with FVIII proteins and/or peptides, consistent with earlier publications from the Conti-Fine group. Peptide ELISPOT assays identified immunogenic regions of FVIII. Specificity for sequences encoded within PBMCs from multiple subjects with an Int22Inv mutation, with and without a current FVIII inhibitor, responded to FVIII epitopes. Furthermore, the FVIII region encoded by the exon 22-23 junction sequence was not remarkably immunoreactive and is therefore unlikely to contain an immunodominant, promiscuous CD4
Sections du résumé
Background
Almost half of severe hemophilia A (HA) is caused by an intron 22 inversion mutation (Int22Inv), which disrupts the 26-exon
Objectives
To test the hypothesis that (putative) intracellular synthesis of FVIII proteins encoded by inverted
Patients/Methods
Peripheral blood mononuclear cells (PBMCs) from 30 severe or moderate HA subjects (17 with an Int22Inv mutation) were tested by ELISPOT assays to detect cytokine secretion in response to FVIII proteins and peptides and to map immunodominant T-cell epitopes. Potential immunogenicity of FVIII sequences encoded by the
Results
Eight of the Int22Inv subjects showed robust cytokine secretion from PBMCs stimulated with FVIII proteins and/or peptides, consistent with earlier publications from the Conti-Fine group. Peptide ELISPOT assays identified immunogenic regions of FVIII. Specificity for sequences encoded within
Conclusions
PBMCs from multiple subjects with an Int22Inv mutation, with and without a current FVIII inhibitor, responded to FVIII epitopes. Furthermore, the FVIII region encoded by the exon 22-23 junction sequence was not remarkably immunoreactive and is therefore unlikely to contain an immunodominant, promiscuous CD4
Identifiants
pubmed: 36936969
doi: 10.3389/fimmu.2023.1128641
pmc: PMC10015889
doi:
Substances chimiques
Factor VIII
9001-27-8
Peptides
0
Epitopes, T-Lymphocyte
0
RNA, Messenger
0
Cytokines
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1128641Subventions
Organisme : NHLBI NIH HHS
ID : R01 HL130448
Pays : United States
Organisme : NHLBI NIH HHS
ID : U34 HL114674
Pays : United States
Informations de copyright
Copyright © 2023 Gunasekera, Vir, Karim, Ragni and Pratt.
Déclaration de conflit d'intérêts
Author KP is an inventor on patents related to FVIII immunogenicity. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Références
Genomics. 1992 Nov;14(3):585-9
pubmed: 1427887
PLoS Comput Biol. 2008 Apr 04;4(4):e1000048
pubmed: 18389056
J Thromb Haemost. 2003 Oct;1(10):2159-66
pubmed: 14521599
Biomed Res Int. 2013;2013:793502
pubmed: 23555096
Nucleic Acids Res. 2020 Jul 2;48(W1):W449-W454
pubmed: 32406916
Blood. 2013 May 16;121(20):4046-55
pubmed: 23553768
PLoS One. 2015 Jan 23;10(1):e0116577
pubmed: 25615825
Haemophilia. 2009 May;15(3):712-7
pubmed: 19432924
Blood. 2003 Feb 15;101(4):1351-8
pubmed: 12393451
Haemophilia. 2010 May;16(102):44-55
pubmed: 20536985
J Thromb Haemost. 2003 Aug;1(8):1777-84
pubmed: 12911593
Nat Genet. 1993 Nov;5(3):236-41
pubmed: 8275087
J Thromb Haemost. 2022 Sep;20(9):2022-2034
pubmed: 35770352
BMC Bioinformatics. 2010 Nov 22;11:568
pubmed: 21092157
Blood. 2009 Aug 13;114(7):1423-8
pubmed: 19549986
Haemophilia. 2009 Sep;15(5):1074-82
pubmed: 19563499
Haemophilia. 2006 Dec;12 Suppl 6:23-8; discussion 28-9
pubmed: 17123390
Nat Med. 2013 Oct;19(10):1318-24
pubmed: 24037092
Haemophilia. 2001 May;7(3):267-72
pubmed: 11380630
Blood. 2015 Jan 8;125(2):223-8
pubmed: 25406352
Blood. 2016 Oct 20;128(16):2043-2054
pubmed: 27471234
Hum Mutat. 2013 Feb;34(2):E2382-91
pubmed: 23280990
Blood. 2015 Aug 13;126(7):895-904
pubmed: 25617427
Semin Thromb Hemost. 2009 Nov;35(8):723-34
pubmed: 20169509
Nature. 1984 Nov 22-28;312(5992):326-30
pubmed: 6438525
Blood. 2012 Mar 22;119(12):2922-34
pubmed: 22282501
Thromb Haemost. 2002 Oct;88(4):568-75
pubmed: 12362225
Blood Adv. 2018 Feb 27;2(4):309-322
pubmed: 29444872
Am J Hematol. 1993 Apr;42(4):375-9
pubmed: 8493988
Eur J Haematol. 2015 Feb;94 Suppl 77:38-44
pubmed: 25560793
J Thromb Haemost. 2007 Dec;5(12):2399-407
pubmed: 18034765
Thromb Haemost. 2007 May;97(5):788-94
pubmed: 17479189
Blood Adv. 2020 Nov 24;4(22):5785-5796
pubmed: 33232473
Hum Mol Genet. 1993 Nov;2(11):1773-8
pubmed: 8281136
J Thromb Haemost. 2004 Aug;2(8):1385-94
pubmed: 15304045
J Thromb Haemost. 2017 Oct;15(10):1971-1976
pubmed: 28795528
Expert Rev Vaccines. 2012 Jan;11(1):43-54
pubmed: 22149708
Thromb Haemost. 1999 Aug;82(2):509-15
pubmed: 10605744
J Thromb Haemost. 2011 Apr;9(4):689-99
pubmed: 21251204
J Thromb Haemost. 2004 Nov;2(11):1908-17
pubmed: 15550021