Dataset Information

Exploring red blood cells as an antigen delivery system to modulate the immune response towards FVIII in hemophilia A

ABSTRACT: Background: The main complication in hemophilia A treatment is the development of inhibitory antibodies against factor (F)VIII. Immune tolerance induction, the gold standard for eradicating anti-FVIII antibodies, is efﬁcient in only 60% to 80% of cases. This underscores the need for more efﬁcient induction of tolerance in patients with hemophilia A with FVIII inhibitors. Objectives: In this study, we explored whether red blood cells (RBCs) can be utilized as antigen delivery system to modulate the immune response against FVIII. Methods: Two promiscuously HLA-DR–presented peptides derived from the A2 and C1 domains of FVIII were fused to the TAT cell-penetrating peptide and incubated with RBCs. Results: Biotinylated TAT-A2 and TAT-C1 peptides were found to interact with RBCs as shown by ﬂow cytometry and imaging ﬂow cytometry. Moreover, macrophages efﬁciently phagocytosed TAT-FVIII peptide–treated RBCs. Using mass spectrometry– based immunopeptidomics we established that TAT-FVIII peptides were presented on major histocompatibility complex class II of macrophages that phagocytosed TAT peptide–pulsed RBCs. Speciﬁcally, the TAT-A2 peptide exhibited efﬁcient processing and presentation on HLA-DR molecules. Importantly, incubation of TAT-C1 peptide– treated RBCs-loaded macrophages with a FVIII-speciﬁc T-cell hybridoma led to a signiﬁcant increase in IL-2 production, suggesting functional presentation of TAT-C1– derived peptides by macrophages. Conclusion: Our ﬁndings indicate that RBCs can serve as effective vehicle for the delivery of FVIII-derived peptides to antigen-presenting cells. The successful display of T-cell epitopes on antigen-presenting cell using ex vivo–loaded RBC may be potentially utilized to modulate pathogenic immune responses such as observed in a subset of patients with hemophilia A.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood

DISEASE(S): Autosomal Hemophilia A

SUBMITTER: Arie Hoogendijk

LAB HEAD: A.J. Hoogendijk

PROVIDER: PXD069058 | Pride | 2025-10-02

REPOSITORIES: Pride

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Exploring red blood cells as an antigen delivery system to modulate the immune response towards FVIII in hemophilia A.

Miranda Mariarosaria M Brandsma Eelke E Robben Lotte L Van Dender Helena H van Alphen Floris P J FPJ Fijnvandraat Karin K van den Biggelaar Maartje M Lacroix-Desmazes Sebastien S van Bruggen Robin R Voorberg Jan J

Journal of thrombosis and haemostasis : JTH 20241129 3

<h4>Background</h4>The main complication in hemophilia A treatment is the development of inhibitory antibodies against factor (F)VIII. Immune tolerance induction, the gold standard for eradicating anti-FVIII antibodies, is efficient in only 60% to 80% of cases. This underscores the need for more efficient induction of tolerance in patients with hemophilia A with FVIII inhibitors.<h4>Objectives</h4>In this study, we explored whether red blood cells (RBCs) can be utilized as antigen delivery syste ...[more]

PMID: 39617188

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Project description:Aims Leveraging the tolerogenic nature of their natural clearance pathway, we aim to exploit red blood cells (RBCs) as an antigen delivery system to achieve persistent exposure to ADAMTS13-derived peptides, promoting antigen-specific attenuation of autoreactive CD4+ T cells and induction of regulatory T cells. Background Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is a rare and potentially fatal autoimmune disease caused by a severe ADAMTS13 functional deficiency mediated by autoantibodies targeting ADAMTS13. Despite high survival rates achieved with current treatments, approximately 40% of patients experience relapses. Ensuring longer-lasting recovery by restoring immune tolerance towards ADAMTS13 remains a significant unmet need. Conclusion(s) Our strategy offers a modular and effective method for targeting antigenic peptides to RBCs for transfusion. Antigen-decorated RBCs are efficiently phagocytosed by macrophages and facilitate the presentation of FINVAPHAR on MHC II molecules, giving macrophages the ability to target ADAMTS13-specific T cells. Based on our results we propose that RBCs loaded with ADAMTS13-derived peptides containing immunodominant T cell epitopes represent a promising strategy for promoting tolerance in patients with iTTP. Methods A fusion peptide comprising the TAT cell-penetrating peptide and an immunodominant ADAMTS13-derived T cell epitope (FINVAPHAR core sequence) was designed. Binding of the fusion peptide to RBCs was monitored by flow cytometry and Imagestream. To examine whether this approach can facilitate antigen-presentation on MHC II molecules, peptide-loaded RBCs were fed to macrophages, followed by isolation of HLA-DR-peptide complexes to study the presented peptide repertoire via mass spectrometry. Results We found that the fusion peptide binds to RBCs in a concentration-dependent manner. Peptide-bound RBCs were efficiently phagocytosed by macrophages. Mass spectrometric analysis revealed that phagocytosis of peptide-loaded RBCs by macrophages results in the presentation of FINVAPHAR-containing sequences of varying lengths on MHCII molecules from HLA-DRB1*11 donors, confirming functional antigen presentation

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	DonorB_MO_RBCs_A2.raw	Raw