NCF4 regulates antigen presentation of cysteine peptides by intracellular oxidative response and restricts activation of autoreactive and arthritogenic T cells
Jing Xu,
Chang He,
Yongsong Cai,
Xipeng Wang,
Jidong Yan,
Jing Zhang,
Fujun Zhang,
Vilma Urbonaviciute,
Yuanyuan Cheng,
Shemin Lu,
Rikard Holmdahl
Affiliations
Jing Xu
National Joint Engineering Research Center of Biodiagnostics and Biotherapy, and Department of Rheumatology, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi, 710004, PR China; Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China; Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China; Medical Inflammation Research, Division of Immunology, Dept. of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
Chang He
Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China; Medical Inflammation Research, Division of Immunology, Dept. of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden; Department of Cardiology, The Second Affiliated Hospital, Zhejiang University Schoole of Medicine, Zhejiang, Hangzhou, PR China
Yongsong Cai
Department of Joint Surgery, Xi'an Honghui Hospital, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China
Xipeng Wang
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China; Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China
Jidong Yan
Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, 710061, Xi'an, PR China
Jing Zhang
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China; Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China
Fujun Zhang
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China; Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China
Vilma Urbonaviciute
Medical Inflammation Research, Division of Immunology, Dept. of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
Yuanyuan Cheng
National Joint Engineering Research Center of Biodiagnostics and Biotherapy, and Department of Rheumatology, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi, 710004, PR China
Shemin Lu
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China; Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China
Rikard Holmdahl
National Joint Engineering Research Center of Biodiagnostics and Biotherapy, and Department of Rheumatology, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi, 710004, PR China; Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, PR China; Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education, Xi'an, Shaanxi, 710061, PR China; Medical Inflammation Research, Division of Immunology, Dept. of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden; Corresponding author. Medical Inflammation Research, Division of Immunology, Dept. of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.
Autoimmune diseases, such as rheumatoid arthritis (RA) and systemic lupus erythematous, are regulated by polymorphisms in genes contributing to the NOX2 complex. Mutations in both Ncf1 and Ncf4 affect development of arthritis in experimental models of RA, but the different regulatory pathways mediated by NOX2-derived reactive oxygen species (ROS) have not yet been clarified.Here we address the possibility that intracellular ROS, regulated by the NCF4 protein (earlier often denoted p40phox) which interacts with endosomal membranes, could play an important role in the oxidation of cysteine peptides in mononuclear phagocytic cells, thereby regulating antigen presentation and activation of arthritogenic T cells.To study the role of NCF4 we used mice with an amino acid replacing mutation (NCF4R58A), which is known to affect interaction with endosomal membranes, leading to decreased intracellular ROS production. To study the impact of NCF4 on T cell activation, we used the glucose phosphate isomerase peptide GPI325-339, which contains two cysteine residues (325-339c-c). Macrophages from mice with the NCF458A mutation efficiently presented the peptide when the two cysteines were intact and not crosslinked, leading to a strong arthritogenic T cell response. T cell priming occurred in the draining lymph nodes (LNs) within 8 days after immunization. Clodronate treatment, which depletes antigen-presenting mononuclear phagocytes, ameliorated arthritis severity, whereas treatment with FYT720, which traps activated T cells in LNs, prohibited arthritis.We conclude that NCF4-dependent intracellular ROS maintains cysteine peptides in an oxidized crosslinked state, which prevents presentation of peptides recognized by non-tolerized T cells and thereby protects against autoimmune arthritis.
