Quantitative HLA-class-II/factor VIII (FVIII) peptidomic variation in dendritic cells correlates with the immunogenic potential of therapeutic FVIII proteins in hemophilia A

Vincent P Diego; Bernadette W Luu; Marco Hofmann; Long V Dinh; Marcio Almeida; Jerry S Powell; Raja Rajalingam; Juan M Peralta; Satish Kumar; Joanne E Curran; Zuben E Sauna; Roberta Kellerman; Yara Park; Nigel S Key; Miguel A Escobar; Huy Huynh; Anne M Verhagen; Sarah Williams-Blangero; Paul V Lehmann; Eugene Maraskovsky; John Blangero; Tom E Howard

doi:10.1111/jth.14647

Quantitative HLA-class-II/factor VIII (FVIII) peptidomic variation in dendritic cells correlates with the immunogenic potential of therapeutic FVIII proteins in hemophilia A

J Thromb Haemost. 2020 Jan;18(1):201-216. doi: 10.1111/jth.14647.

Authors

Vincent P Diego^{1

2}, Bernadette W Luu^{1

2

3}, Marco Hofmann⁴, Long V Dinh³, Marcio Almeida^{1

2}, Jerry S Powell⁵, Raja Rajalingam⁶, Juan M Peralta^{1

2}, Satish Kumar^{1

2}, Joanne E Curran^{1

2}, Zuben E Sauna⁷, Roberta Kellerman⁸, Yara Park⁹, Nigel S Key^{8

9}, Miguel A Escobar¹⁰, Huy Huynh¹¹, Anne M Verhagen¹¹, Sarah Williams-Blangero^{1

2}, Paul V Lehmann^{12

13

14}, Eugene Maraskovsky¹¹, John Blangero^{1

2}, Tom E Howard^{1

2

3

15}

Affiliations

¹ South Texas Diabetes and Obesity Institute, University of Texas Rio Grande Valley, Brownsville, Texas.
² Department of Human Genetics, School of Medicine, University of Texas Rio Grande Valley, Brownsville, Texas.
³ Haplogenics Corporation, Brownsville, Texas.
⁴ CSL Behring GmbH, Research, Marburg, Germany.
⁵ CSL Behring, Med Affairs, King of Prussia, Pennsylvania.
⁶ Immunogenetics and Transplantation Laboratory, Department of Surgery, School of Medicine, University of California at San Francisco, California.
⁷ Hemostasis Branch, Division of Plasma Protein Therapeutics, Office of Tissues and Advanced Therapeutics, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland.
⁸ Division of Hematology, Department of Medicine, University of North Carolina at Chapel Hill, North Carolina.
⁹ Department of Laboratory Medicine and Pathology, University of North Carolina at Chapel Hill, North Carolina.
¹⁰ Division of Hematology, Department of Medicine, McGovern School of Medicine, University of Texas Health Sciences Center at Houston, Texas.
¹¹ CSL Limited Research, Bio21 Institute, Melbourne, Australia.
¹² Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio.
¹³ Department of Neurology, Case Western Reserve University School of Medicine, Cleveland, Ohio.
¹⁴ Cellular Technology Ltd, Shaker Heights, Ohio.
¹⁵ Department of Pathology and Lab Medicine, VA Valley Coastal Bend Healthcare Center, Harlingen, Texas.

PMID: 31556206
DOI: 10.1111/jth.14647

Abstract

Background: Plasma-derived (pd) or recombinant (r) therapeutic factor VIII proteins (FVIIIs) are infused to arrest/prevent bleeding in patients with hemophilia A (PWHA). However, FVIIIs are neutralized if anti-FVIII-antibodies (inhibitors) develop. Accumulating evidence suggests that pdFVIIIs with von Willebrand factor (VWF) are less immunogenic than rFVIIIs and that distinct rFVIIIs are differentially immunogenic. Since inhibitor development is T-helper-cell-dependent, human leukocyte antigen (HLA)-class-II (HLAcII) molecules constitute an important early determinant.

Objectives: Use dendritic cell (DC)-protein processing/presentation assays with mass-spectrometric and peptide-proteomic analyses to quantify the DP-bound, DQ-bound, and DR-bound FVIII-derived peptides in individual HLAcII repertoires and compare the immunogenic potential of six distinct FVIIIs based on their measured peptide counts.

Patients/methods: Monocyte-derived DCs from normal donors and/or PWHA were cultured with either: Mix-rFVIII, a VWF-free equimolar mixture of a full-length (FL)-rFVIII [Advate^® (Takeda)] and four distinct B-domain-deleted (BDD)-rFVIIIs [Xyntha^® (Pfizer), NovoEight^® (Novo-Nordisk), Nuwiq^® (Octapharma), and Afstyla^® (CSL Behring GmBH)]; a pdFVIII + pdVWF [Beriate^® (CSL Behring GmBH)]; Advate ± pdVWF; Afstyla ± pdVWF; and Xyntha + pdVWF.

Results: We showed that (i) Beriate had a significantly lower immunogenic potential than Advate ± pdVWF, Afstyla - pdVWF, and Mix-rFVIII; (ii) distinct FVIIIs differed significantly in their immunogenic potential in that, in addition to (i), Afstyla + pdVWF had a significantly lower immunogenic potential than Beriate, while the immunogenic potential of Beriate was not significantly different from that of Xyntha + pdVWF; and (iii) rFVIIIs with pdVWF had significantly lower immunogenic potentials than the same rFVIIIs without pdVWF.

Conclusions: Our results provide HLAcII peptidomic level explanations for several important clinical observations/issues including the differential immunogenicity of distinct FVIIIs and the role of HLAcII genetics in inhibitor development.

Keywords: blood coagulation factor inhibitors; factor VIII; hemophilia A; histocompatibility antigens class II; peptide; quantitative peptidomics; sequence analysis.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Dendritic Cells
Factor VIII*
HLA Antigens
Hemophilia A* / drug therapy
Humans
Proteomics

Substances

HLA Antigens
F8 protein, human
Factor VIII

Abstract

Publication types

MeSH terms

Substances

Grants and funding