Cloé Comarmond1, Valérie Lorin2, Cindy Marques3, Anna Maciejewski-Duval4, Nizar Joher2, Cyril Planchais2, Maxime Touzot5, Lucie Biard6, Thierry Hieu2, Valentin Quiniou7, Anne-Claire Desbois4, Michelle Rosenzwajg7, David Klatzmann4, Patrice Cacoub4, Hugo Mouquet8, David Saadoun9. 1. Sorbonne Université, INSERM UMR_S 959, Immunologie-Immunopathologie-Immunotherapie, i3 and Département Hospitalo-Universitaire Inflammation-Immunopathologie-Biotherapie, i2B, F-75651 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service de Biothérapie, F-75013 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Département de Médecine Interne et Immunologie Clinique, F-75013 Paris, France; Laboratory of Humoral Immunology, Department of Immunology, Institut Pasteur, Paris, France; INSERM U1222, Paris, France. 2. Laboratory of Humoral Immunology, Department of Immunology, Institut Pasteur, Paris, France; INSERM U1222, Paris, France. 3. AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Département de Médecine Interne et Immunologie Clinique, F-75013 Paris, France; Laboratory of Humoral Immunology, Department of Immunology, Institut Pasteur, Paris, France; INSERM U1222, Paris, France. 4. Sorbonne Université, INSERM UMR_S 959, Immunologie-Immunopathologie-Immunotherapie, i3 and Département Hospitalo-Universitaire Inflammation-Immunopathologie-Biotherapie, i2B, F-75651 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service de Biothérapie, F-75013 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Département de Médecine Interne et Immunologie Clinique, F-75013 Paris, France. 5. INSERM U932, 26 rue d'Ulm, 75005 Paris, France; Institut Curie, Section Recherche, 26 rue d'Ulm, 75005 Paris, France; Laboratoire d'Immunologie Clinique, Institut Curie, 26 rue d'Ulm, 75005 Paris, France. 6. AP-HP, SBIM, Hôpital Saint-Louis, Université Paris Diderot, Paris 7, Paris, France; INSERM, ECSTRA Team, CRESS UMR-S 1153, 75010 Paris, France. 7. Sorbonne Université, INSERM UMR_S 959, Immunologie-Immunopathologie-Immunotherapie, i3 and Département Hospitalo-Universitaire Inflammation-Immunopathologie-Biotherapie, i2B, F-75651 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service de Biothérapie, F-75013 Paris, France. 8. Laboratory of Humoral Immunology, Department of Immunology, Institut Pasteur, Paris, France; INSERM U1222, Paris, France. Electronic address: hugo.mouquet@pasteur.fr. 9. Sorbonne Université, INSERM UMR_S 959, Immunologie-Immunopathologie-Immunotherapie, i3 and Département Hospitalo-Universitaire Inflammation-Immunopathologie-Biotherapie, i2B, F-75651 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service de Biothérapie, F-75013 Paris, France; AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Département de Médecine Interne et Immunologie Clinique, F-75013 Paris, France. Electronic address: david.saadoun@aphp.fr.
Abstract
BACKGROUND & AIMS: Hepatitis C virus (HCV) infection contributes to the development of autoimmune disorders such as cryoglobulinaemia vasculitis (CV). However, it remains unclear why only some individuals with HCV develop HCV-associated CV (HCV-CV). HCV-CV is characterized by the expansion of anergic CD19+CD27+CD21low/- atypical memory B cells (AtMs). Herein, we report the mechanisms by which AtMs participate in HCV-associated autoimmunity. METHODS: The phenotype and function of peripheral AtMs were studied by multicolour flow cytometry and co-culture assays with effector T cells and regulatory T cells in 20 patients with HCV-CV, 10 chronicallyHCV-infected patients without CV and 8 healthy donors. We performed gene expression profile analysis of AtMs stimulated or not by TLR9. Immunoglobulin gene repertoire and antibody reactivity profiles of AtM-expressing IgM antibodies were analysed following single B cell FACS sorting and expression-cloning of monoclonal antibodies. RESULTS: The Tbet+CD11c+CD27+CD21- AtM population is expanded in patients with HCV-CV compared to HCV controls without CV. TLR9 activation of AtMs induces a specific transcriptional signature centred on TNFα overexpression, and an enhanced secretion of TNFα and rheumatoid factor-type IgMs in patients with HCV-CV. AtMs stimulated through TLR9 promote type 1 effector T cell activation and reduce the proliferation of CD4+CD25hiCD127-/lowFoxP3+ regulatory T cells. AtM expansions display intraclonal diversity with immunoglobulin features of antigen-driven maturation. AtM-derived IgM monoclonal antibodies do not react against ubiquitous autoantigens or HCV antigens including NS3 and E2 proteins. Rather, AtM-derived antibodies possess rheumatoid factor activity and target unique epitopes on the human IgG-Fc region. CONCLUSION: Our data strongly suggest a central role for TLR9 activation of AtMs in driving HCV-CV autoimmunity through rheumatoid factor production and type 1 T cell responses. LAY SUMMARY: B cells are best known for their capacity to produce antibodies, which often play a deleterious role in the development of autoimmune diseases. During chronic hepatitis C, self-reactive B cells proliferate and can be responsible for autoimmune symptoms (arthritis, purpura, neuropathy, renal disease) and/or lymphoma. Direct-acting antiviral therapy clears the hepatitis C virus and eliminates deleterious B cells.
BACKGROUND & AIMS:Hepatitis C virus (HCV) infection contributes to the development of autoimmune disorders such as cryoglobulinaemia vasculitis (CV). However, it remains unclear why only some individuals with HCV develop HCV-associated CV (HCV-CV). HCV-CV is characterized by the expansion of anergic CD19+CD27+CD21low/- atypical memory B cells (AtMs). Herein, we report the mechanisms by which AtMs participate in HCV-associated autoimmunity. METHODS: The phenotype and function of peripheral AtMs were studied by multicolour flow cytometry and co-culture assays with effector T cells and regulatory T cells in 20 patients with HCV-CV, 10 chronicallyHCV-infected patients without CV and 8 healthy donors. We performed gene expression profile analysis of AtMs stimulated or not by TLR9. Immunoglobulin gene repertoire and antibody reactivity profiles of AtM-expressing IgM antibodies were analysed following single B cell FACS sorting and expression-cloning of monoclonal antibodies. RESULTS: The Tbet+CD11c+CD27+CD21- AtM population is expanded in patients with HCV-CV compared to HCV controls without CV. TLR9 activation of AtMs induces a specific transcriptional signature centred on TNFα overexpression, and an enhanced secretion of TNFα and rheumatoid factor-type IgMs in patients with HCV-CV. AtMs stimulated through TLR9 promote type 1 effector T cell activation and reduce the proliferation of CD4+CD25hiCD127-/lowFoxP3+ regulatory T cells. AtM expansions display intraclonal diversity with immunoglobulin features of antigen-driven maturation. AtM-derived IgM monoclonal antibodies do not react against ubiquitous autoantigens or HCV antigens including NS3 and E2 proteins. Rather, AtM-derived antibodies possess rheumatoid factor activity and target unique epitopes on the human IgG-Fc region. CONCLUSION: Our data strongly suggest a central role for TLR9 activation of AtMs in driving HCV-CV autoimmunity through rheumatoid factor production and type 1 T cell responses. LAY SUMMARY: B cells are best known for their capacity to produce antibodies, which often play a deleterious role in the development of autoimmune diseases. During chronic hepatitis C, self-reactive B cells proliferate and can be responsible for autoimmune symptoms (arthritis, purpura, neuropathy, renal disease) and/or lymphoma. Direct-acting antiviral therapy clears the hepatitis C virus and eliminates deleterious B cells.
Authors: Swati Phalke; Katja Aviszus; Kira Rubtsova; Anatoly Rubtsov; Briana Barkes; Linda Powers; Brenda Warner; James L Crooks; John W Kappler; Evans R Fernández-Pérez; Lisa A Maier; Nabeel Hamzeh; Philippa Marrack Journal: Am J Respir Crit Care Med Date: 2020-10-01 Impact factor: 21.405
Authors: Bruna Kulmann-Leal; Joel Henrique Ellwanger; Jacqueline María Valverde-Villegas; Daniel Simon; Camila Guerra Marangon; Vanessa Suñé Mattevi; Rosmeri Kuhmmer Lazzaretti; Regina Kuhmmer; Eduardo Sprinz; José Artur Bogo Chies Journal: Genet Test Mol Biomarkers Date: 2022-05-16