Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators
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 th...
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| Format: | Article |
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Frontiers Media S.A.
2020-06-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.01219/full |
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| author | Ahmad Faisal Karim Ahmad Faisal Karim Anthony R. Soltis Anthony R. Soltis Anthony R. Soltis Gauthaman Sukumar Gauthaman Sukumar Gauthaman Sukumar Christoph Königs Nadia P. Ewing Clifton L. Dalgard Clifton L. Dalgard Clifton L. Dalgard Matthew D. Wilkerson Matthew D. Wilkerson Matthew D. Wilkerson Matthew D. Wilkerson Kathleen P. Pratt |
| author_facet | Ahmad Faisal Karim Ahmad Faisal Karim Anthony R. Soltis Anthony R. Soltis Anthony R. Soltis Gauthaman Sukumar Gauthaman Sukumar Gauthaman Sukumar Christoph Königs Nadia P. Ewing Clifton L. Dalgard Clifton L. Dalgard Clifton L. Dalgard Matthew D. Wilkerson Matthew D. Wilkerson Matthew D. Wilkerson Matthew D. Wilkerson Kathleen P. Pratt |
| author_sort | Ahmad Faisal Karim |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T22:37:03Z |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-11T22:37:03Z |
| publishDate | 2020-06-01 |
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| spelling | doaj.art-82e845a0c3b84010b757baaccfdac88b2022-12-22T00:47:55ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-06-011110.3389/fimmu.2020.01219529322Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune ModulatorsAhmad Faisal Karim0Ahmad Faisal Karim1Anthony R. Soltis2Anthony R. Soltis3Anthony R. Soltis4Gauthaman Sukumar5Gauthaman Sukumar6Gauthaman Sukumar7Christoph Königs8Nadia P. Ewing9Clifton L. Dalgard10Clifton L. Dalgard11Clifton L. Dalgard12Matthew D. Wilkerson13Matthew D. Wilkerson14Matthew D. Wilkerson15Matthew D. Wilkerson16Kathleen P. Pratt17Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesHenry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United StatesHenry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United StatesCollaborative Health Initiative Research Program, Henry Jackson Foundation for the Advancement of Military Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesThe American Genome Center, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesCollaborative Health Initiative Research Program, Henry Jackson Foundation for the Advancement of Military Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesThe American Genome Center, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesDepartment of Anatomy, Physiology & Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesDepartment of Pediatrics, Goethe University, Frankfurt, GermanyCity of Hope National Medical Center, Duarte, CA, United StatesCollaborative Health Initiative Research Program, Henry Jackson Foundation for the Advancement of Military Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesThe American Genome Center, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesDepartment of Anatomy, Physiology & Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesHenry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United StatesCollaborative Health Initiative Research Program, Henry Jackson Foundation for the Advancement of Military Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesThe American Genome Center, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesDepartment of Anatomy, Physiology & Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesDepartment of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United StatesFormation 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.https://www.frontiersin.org/article/10.3389/fimmu.2020.01219/fullhemophilia ARNAseq analysisinnate and adaptive immune responsefactor VIII (FVIII)PBMC (peripheral blood mononuclear cells) |
| spellingShingle | Ahmad Faisal Karim Ahmad Faisal Karim Anthony R. Soltis Anthony R. Soltis Anthony R. Soltis Gauthaman Sukumar Gauthaman Sukumar Gauthaman Sukumar Christoph Königs Nadia P. Ewing Clifton L. Dalgard Clifton L. Dalgard Clifton L. Dalgard Matthew D. Wilkerson Matthew D. Wilkerson Matthew D. Wilkerson Matthew D. Wilkerson Kathleen P. Pratt Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators Frontiers in Immunology hemophilia A RNAseq analysis innate and adaptive immune response factor VIII (FVIII) PBMC (peripheral blood mononuclear cells) |
| title | Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators |
| title_full | Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators |
| title_fullStr | Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators |
| title_full_unstemmed | Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators |
| title_short | Hemophilia A Inhibitor Subjects Show Unique PBMC Gene Expression Profiles That Include Up-Regulated Innate Immune Modulators |
| title_sort | hemophilia a inhibitor subjects show unique pbmc gene expression profiles that include up regulated innate immune modulators |
| topic | hemophilia A RNAseq analysis innate and adaptive immune response factor VIII (FVIII) PBMC (peripheral blood mononuclear cells) |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2020.01219/full |
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