Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators
Open Access
- 12 June 2020
- journal article
- research article
- Published by Frontiers Media SA in Frontiers in Immunology
- Vol. 11, 1219
- https://doi.org/10.3389/fimmu.2020.01219
Abstract
Formation of pathological anti-FVIII antibodies, or “inhibitors,” is the most serious complication of therapeutic FVIII infusions, affecting up to 1/3 of severe Hemophilia A (HA) patients. Inhibitor formation is a classical T-cell dependent adaptive immune response. As such, it requires help from the innate immune system. However, the roles of innate immune cells and mechanisms of inhibitor development vs. immune tolerance, achieved with or without Immune Tolerance Induction (ITI) therapy, are not well-understood. To address these questions, temporal transcriptomics profiling of FVIII-stimulated peripheral blood mononuclear cells (PBMCs) was carried out for HA subjects with and without a current or historic inhibitor using RNA-Seq. PBMCs were isolated from 40 subjects in the following groups: HA with an inhibitor that resolved either following ITI or spontaneously; HA with a current inhibitor; HA with no inhibitor history and non-HA controls. PBMCs were stimulated with 5 nM FVIII and RNA was isolated 4, 16, 24, and 48 h following stimulation. Time-series differential expression analysis was performed and distinct transcriptional signatures were identified for each group, providing clues as to cellular mechanisms leading to or accompanying their disparate anti-FVIII antibody responses. Subjects with a current inhibitor showed differential expression of 56 genes and a clustering analysis identified three major temporal profiles. Interestingly, gene ontology enrichments featured innate immune modulators, including NLRP3, TLR8, IL32, CLEC10A, and COLEC12. NLRP3 and TLR8 are associated with enhanced secretion of the pro-inflammatory cytokines IL-1β and TNFα, while IL32, which has several isoforms, has been associated with both inflammatory and regulatory immune processes. RNA-Seq results were validated by RT-qPCR, ELISAs, multiplex cytokine analysis, and flow cytometry. The inflammatory status of HA patients suffering from an ongoing inhibitor includes up-regulated innate immune modulators, which may act as ongoing danger signals that influence the responses to, and eventual outcomes of, ITI therapy.Funding Information
- National Heart, Lung, and Blood Institute
This publication has 64 references indexed in Scilit:
- MicroRNAs bind to Toll-like receptors to induce prometastatic inflammatory responseProceedings of the National Academy of Sciences of the United States of America, 2012
- HLA-DR-presented Peptide Repertoires Derived From Human Monocyte-derived Dendritic Cells Pulsed With Blood Coagulation Factor VIIIMolecular & Cellular Proteomics, 2011
- Effect of B-cell depletion using anti-CD20 therapy on inhibitory antibody formation to human FVIII in hemophilia A miceBlood, 2011
- TMEPAI, a Transmembrane TGF-β-Inducible Protein, Sequesters Smad Proteins from Active Participation in TGF-β SignalingMolecular Cell, 2010
- IL‐1β, IL‐6, KC and MCP‐1 are elevated in synovial fluid from haemophilic mice with experimentally induced haemarthrosisHaemophilia, 2009
- Recombinant factor VIII and factor VIII-von Willebrand factor complex do not present danger signals for human dendritic cellsThrombosis and Haemostasis, 2006
- Interleukin-32: A Cytokine and Inducer of TNFαImmunity, 2005
- Immunoregulatory role of interleukin 10 in rheumatoid arthritis.The Journal of Experimental Medicine, 1994
- Loss of high‐responder inhibitors in patients with severe hemophilia A and human immunodeficiency virus type 1 infection: A report from the multi‐center hemophilia cohort studyAmerican Journal of Hematology, 1993
- Disappearance of inhibitor to factor VIII in HIV-infected hemophiliacs with progression to AIDS or severe ARCTransfusion, 1989