Helminth‐induced CD19+CD23hi B cells modulate experimental allergic and autoimmune inflammation

Numerous population studies and experimental models suggest that helminth infections can ameliorate immuno‐inflammatory disorders such as asthma and autoimmunity. Immunosuppressive cell populations associated with helminth infections include Treg and alternatively‐activated macrophages. In previous studies, we showed that both CD4⁺CD25⁺ Treg, and CD4^– MLN cells from Heligmosomoides polygyus‐infected C57BL/6 mice were able to transfer protection against allergic airway inflammation to sensitized but uninfected animals. We now show that CD4^–CD19⁺ MLN B cells from infected, but not naïve, mice are able to transfer a down‐modulatory effect on allergy, significantly suppressing airway eosinophilia, IL‐5 secretion and pathology following allergen challenge. We further demonstrate that the same cell population can alleviate autoimmune‐mediated inflammatory events in the CNS, when transferred to uninfected mice undergoing myelin oligodendrocyte glycoprotein_(p35–55)‐induced EAE. In both allergic and autoimmune models, reduction of disease was achieved with B cells from helminth‐infected IL‐10^?/? donors, indicating that donor cell‐derived IL‐10 is not required. Phenotypically, MLN B cells from helminth‐infected mice expressed uniformly high levels of CD23, with follicular (B2) cell surface markers. These data expand previous observations and highlight the broad regulatory environment that develops during helminth infections that can abate diverse inflammatory disorders in vivo.     

IL‐17‐producing CD4+ T cells contribute to the loss of B‐cell tolerance in experimental autoimmune myasthenia gravis

The role of Th17 cells in the pathogenesis of autoantibody‐mediated diseases is unclear. Here, we assessed the contribution of Th17 cells to the pathogenesis of experimental autoimmune myasthenia gravis (EAMG), which is induced by repetitive immunizations with Torpedo californica acetylcholine receptor (tAChR). We show that a significant fraction of tAChR‐specific CD4⁺ T cells is producing IL‐17. IL‐17^ko mice developed fewer or no EAMG symptoms, although the frequencies of tAChR‐specific CD4⁺ T cells secreting IL‐2, IFN‐γ, or IL‐21, and the percentage of FoxP3⁺ T_reg cells were similar to WT mice. Even though the total anti‐tAChR antibody levels were equal, the complement fixating IgG2b subtype was reduced in IL‐17^ko as compared to WT mice. Most importantly, pathogenic anti‐murine AChR antibodies were significantly lower in IL‐17^ko mice. Furthermore, we confirmed the role of Th17 cells in EAMG pathogenesis by the reconstitution of TCR β/δ^ko mice with WT or IL‐17^ko CD4⁺ T cells. In conclusion, we show that the level of IgG2b and the loss of B‐cell tolerance, which results in pathogenic anti‐murine AChR‐specific antibodies, are dependent on IL‐17 production by CD4⁺ T cells. Thus, we describe here for the first time how Th17 cells are involved in the induction of classical antibody‐mediated autoimmunity.     

Ligation of TLR7 on CD19+CD1dhi B cells suppresses allergic lung inflammation via regulatory T cells

B cells have been described as having the capacity to regulate cellular immune responses and suppress inflammatory processes. One such regulatory B‐cell population is defined as IL‐10‐producing CD19⁺CD1d^hi cells. Previous work has identified an expansion of these cells in mice infected with the helminth, Schistosoma mansoni. Here, microarray analysis of CD19⁺CD1d^hi B cells from mice infected with S. mansoni demonstrated significantly increased Tlr7 expression, while CD19⁺CD1d^hi B cells from uninfected mice also demonstrated elevated Tlr7 expression. Using IL‐10 reporter, Il10^?/? and Tlr7^?/‐ mice, we formally demonstrate that TLR7 ligation of CD19⁺CD1d^hi B cells increases their capacity to produce IL‐10. In a mouse model of allergic lung inflammation, the adoptive transfer of TLR7‐elicited CD19⁺CD1d^hi B cells reduced airway inflammation and associated airway hyperresponsiveness. Using DEREG mice to deplete FoxP3⁺ T regulatory cells in allergen‐sensitized mice, we show that that TLR7‐elicited CD19⁺CD1d^hi B cells suppress airway hyperresponsiveness via a T regulatory cell dependent mechanism. These studies identify that TLR7 stimulation leads to the expansion of IL‐10‐producing CD19⁺CD1d^hi B cells, which can suppress allergic lung inflammation via T regulatory cells.     

IL‐10 produced by activated human B cells regulates CD4+ T‐cell activation in vitro

IL‐10‐producing regulatory B cells have been identified in mice and shown to downregulate inflammation, making them potentially important for maintenance of tolerance. In this study, we isolated B cells from human blood and spleen, and showed that after a short period of ex vivo stimulation a number of these cells produced IL‐10. The IL‐10‐producing B cells did not fall within a single clearly defined subpopulation, but were enriched in both the memory (CD27⁺) and the transitional (CD38^high) B‐cell compartments. Combined CpG‐B+anti‐Ig stimulation was the most potent IL‐10 stimulus tested. B cells stimulated in this way inhibited CD4⁺CD25^? T‐cell proliferation in vitro by a partially IL‐10‐dependent mechanism. These findings imply that manipulating IL‐10 production by human B cells could be a useful therapeutic strategy for modulating immune responses in humans.     

Expansion of regulatory T cells via IL‐2/anti‐IL‐2 mAb complexes suppresses experimental myasthenia

Human autoimmune diseases are often characterized by a relative deficiency in CD4⁺CD25⁺ regulatory T cells (Treg). We therefore hypothesized that expansion of Treg can ameliorate autoimmune pathology. We tested this hypothesis in an experimental model for autoimmune myasthenia gravis (MG), a B‐cell‐mediated disease characterized by auto‐Ab directed against the acetylcholine receptor within neuromuscular junctions. We showed that injection of immune complexes composed of the cytokine IL‐2 and anti‐IL‐2 mAb (JES6‐1A12) induced an effective and sustained expansion of Treg, via peripheral proliferation of CD4⁺CD25⁺Foxp3⁺ cells and peripheral conversion of CD4⁺CD25^?Foxp3^? cells. The expanded Treg potently suppressed autoreactive T‐ and B‐cell responses to acetylcholine receptor and attenuated the muscular weakness that is characteristic of MG. Thus, IL‐2/anti‐IL‐2 mAb complexes can expand functional Treg in vivo, providing a potential clinical application of this modality for treatment of MG and other autoimmune disorders.     

Pathogenic IL‐23 signaling is required to initiate GM‐CSF‐driven autoimmune myocarditis in mice

Using a mouse model of experimental autoimmune myocarditis (EAM), we showed for the first time that IL‐23 stimulation of CD4⁺ T cells is required only briefly at the initiation of GM‐CFS‐dependent cardiac autoimmunity. IL‐23 signal, acting as a switch, turns on pathogenicity of CD4⁺ T cells, and becomes dispensable once autoreactivity is established. Il23a^?/? mice failed to mount an efficient Th17 response to immunization, and were protected from myocarditis. However, remarkably, transient IL‐23 stimulation ex vivo fully restored pathogenicity in otherwise nonpathogenic CD4⁺ T cells raised from Il23a^?/? donors. Thus, IL‐23 may no longer be necessary to uphold inflammation in established autoimmune diseases. In addition, we demonstrated that IL‐23‐induced GM‐CSF mediates the pathogenicity of CD4⁺ T cells in EAM. The neutralization of GM‐CSF abrogated cardiac inflammation. However, sustained IL‐23 signaling is required to maintain IL‐17A production in CD4⁺ T cells. Despite inducing inflammation in Il23a^?/? recipients comparable to wild‐type (WT), autoreactive CD4⁺ T cells downregulated IL‐17A production without persistent IL‐23 signaling. This divergence on the controls of GM‐CSF‐dependent pathogenicity on one side and IL‐17A production on the other side may contribute to the discrepant efficacies of anti‐IL‐23 therapy in different autoimmune diseases.     

IL‐21 promotes the production of anti‐DNA IgG but is dispensable for kidney damage in lyn−/− mice

The autoimmune disease systemic lupus erythematosus is characterized by loss of tolerance to nuclear Ags and a heightened inflammatory environment, which together result in end organ damage. Lyn‐deficient mice, a model of systemic lupus erythematosus, lack an inhibitor of B‐cell and myeloid cell activation. This results in B‐cell hyper‐responsiveness, plasma cell accumulation, autoantibodies, and glomerulonephritis (GN). IL‐21 is associated with autoimmunity in mice and humans and promotes B‐cell differentiation and class switching. Here, we explore the role of IL‐21 in the autoimmune phenotypes of lyn^–/– mice. We find that IL‐21 mRNA is reduced in the spleens of lyn^–/–IL‐6^–/– and lyn^–/–Btk^lo mice, neither of which produce pathogenic autoantibodies or develop significant GN. While IL‐21 is dispensable for plasma cell accumulation and IgM autoantibodies in lyn^–/– mice, it is required for anti‐DNA IgG antibodies and some aspects of T‐cell activation. Surprisingly, GN still develops in lyn^–/–IL‐21^–/– mice. This likely results from the presence of IgG autoantibodies against a limited set of non‐DNA Ags. These studies identify a specific role for IL‐21 in the class switching of anti‐DNA B cells and demonstrate that neither IL‐21 nor anti‐DNA IgG is required for kidney damage in lyn^–/– mice.     

Role of IL‐10‐producing regulatory B cells in control of cerebral malaria in Plasmodium berghei infected mice

Cerebral malaria (CM) is a neurological syndrome often occurring in severe malaria. Although CM is known as an immunopathology in brain tissue mediated by excessive proinflammatory cytokines, the immunoregulatory mechanism is poorly understood. Here, we investigated the role of IL‐10‐producing regulatory B (Breg) cells in modulating CM development in a murine model of Plasmodium berghei ANKA infection. We observed that blood‐stage P. berghei induced expansion of IL‐10‐producing Breg cells in C57BL/6 mice. Adoptive transfer of IL‐10⁺ Breg cells to P. berghei infected mice significantly reduced the accumulation of NK and CD8⁺ T cells and hemorrhage in brain tissue, and improved the survival of the mice compared with control groups, although parasitemia levels were not altered. Treatment of Breg‐cell recipient mice with anti‐IL‐10 receptor mAb blocked the protective effect of Breg cells. Adoptive transfer of CD4⁺CD25⁺ Treg cells failed to prevent CM in infected mice. Spleen cells from Breg‐cell recipient mice produced increased levels of IL‐10 in vitro. Cell co‐culture showed that purified IL‐10⁺ B cells, but not IL‐10^? B cells, promoted IL‐10 production by CD4⁺ T cells. These results demonstrate that IL‐10‐producing Breg cells may represent an important mechanism for controlling the immunopathology and prevention of CM associated with P. berghei infection.     

The regulation and activation of lupus-associated B cells

Summary: Anti‐double‐stranded DNA (anti‐dsDNA) B cells are regulated in non‐autoimmune mice. While some are deleted or undergo receptor editing, a population of anti‐dsDNA (VH3H9/Vλ1) B cells that emigrate into the periphery has also been identified. These cells have an altered phenotype relative to normal B cells in that they have a reduced lifespan, appear developmentally arrested, and localize primarily to the T/B‐cell interface in the spleen. This phenotype may be the consequence of immature B cells encountering antigen in the absence of T‐cell help. When provided with T‐cell help, the anti‐dsDNA B cells differentiate into antibody‐forming cells. In the context of the autoimmune‐prone lpr/lpr or gld/gld mutations, the VH3H9/Vλ1 anti‐dsDNA B cells populate the B‐cell follicle and by 12 weeks of age produce serum autoantibodies. The early event of anti‐dsDNA B‐cell follicular entry, in the absence of autoantibody production, is dependent upon CD4⁺ T cells. We hypothesize that control of autoantibody production in young autoimmune‐prone mice may be regulated by the counterbalancing effect of T‐regulatory (T_reg) cells. Consistent with this model, we have demonstrated that T_reg cells are able to prevent autoantibody production induced by T‐cell help. Additional studies are aimed at investigating the mechanisms of this suppression as well as probing the impact of distinct forms of T‐cell‐dependent and ‐independent activation on anti‐dsDNA B cells.     

Differential Cytokine Expression and Regulatory Cells in Patients with Primary and Secondary Sjögren's Syndrome

Sjögren's syndrome (SS) is an autoimmune disease characterized by lymphocytic infiltration of the salivary and lacrimal glands. The aim of the study was to characterize and compare the presence of diverse cytokines and regulatory T and B cells in lip minor salivary gland (MSG) biopsies from patients with primary Sjögren's syndrome (pSS), secondary SS (sSS), and patients with connective tissue disease (CTD) without (w/o) SS. We included samples of MSG from 15 pSS, 24 sSS (six scleroderma, nine rheumatoid arthritis and nine lupus patients) and 15 patients with CTD w/o SS. Tissues were examined by an indirect immunoperoxidase technique (goat polyclonal anti‐human IL‐19, goat polyclonal anti‐human IL‐22 or mouse monoclonal anti‐human IL‐24). To determine the subpopulation of CD4⁺/IL‐17A⁺‐, CD4⁺/IL‐4⁺‐, CD4⁺/IFN‐?⁺‐expressing T cells, CD25⁺/Foxp3⁺ Treg cells and CD20⁺/IL‐10⁺‐producing B cell subset, a double‐staining procedure was performed. We estimated the mean percentage of positively staining cells in two fields per sample. CD4⁺/IFN‐?⁺, CD4⁺/IL‐4⁺ and IL‐22⁺ cell percentages were elevated in both SS varieties; however, the cells were more prevalent in pSS. Patients with pSS had a high number of CD4⁺/IL‐17A⁺ and IL‐19⁺ T cells and a lower percentage of IL‐24⁺ cells (P < 0.05). The Treg and IL‐10‐producing B cells were increased in pSS (P < 0.05). Concluding, in our patients, a pro‐inflammatory and regulatory balance coexists in SS, being both responses more intense in pSS. The explanation of these differences may be related to disease activity, disease duration and treatment.     

Regulatory functions of B cells in allergic diseases

B cells are essentially described for their capacity to produce antibodies ensuring anti‐infectious immunity or deleterious responses in the case of autoimmunity or allergy. However, abundant data described their ability to restrain inflammation by diverse mechanisms. In allergy, some regulatory B‐cell subsets producing IL‐10 have been recently described as potent suppressive cells able to restrain inflammatory responses both in vitro and in vivo by regulatory T‐cell differentiation or directly inhibiting T‐cell‐mediated inflammation. A specific deficit in regulatory B cells participates to more severe allergic inflammation. Induction of allergen tolerance through specific immunotherapy induces a specific expansion of these cells supporting their role in establishment of allergen tolerance. However, the regulatory functions carried out by B cells are not exclusively IL‐10 dependent. Indeed, other regulatory mechanisms mediated by B cells are (i) the production of TGF‐β, (ii) the promotion of T‐cell apoptosis by Fas–Fas ligand or granzyme‐B pathways, and (iii) their capacity to produce inhibitory IgG4 and sialylated IgG able to mediate anti‐inflammatory mechanisms. This points to Bregs as interesting targets for the development of new therapies to induce allergen tolerance. In this review, we highlight advances in the study of regulatory mechanisms mediated by B cells and outline what is known about their phenotype as well as their suppressive role in allergy from studies in both mice and humans.     

Interleukin‐2 treatment reverses effects of cAMP‐responsive element modulator α‐over‐expressing T cells in autoimmune‐prone mice

Systemic autoimmune diseases, such as systemic lupus erythematosus (SLE), are often characterized by a failure of self‐tolerance and result in an uncontrolled activation of B cells and effector T cells. Interleukin (IL)‐2 critically maintains homeostasis of regulatory T cells (T_reg) and effector T cells in the periphery. Previously, we identified the cAMP‐responsive element modulator α (CREMα) as a major factor responsible for decreased IL‐2 production in T cells from SLE patients. Additionally, using a transgenic mouse that specifically over‐expresses CREMα in T cells (CD2CREMαtg), we provided in‐vivo evidence that CREMα indeed suppresses IL‐2 production. To analyse the effects of CREMα in an autoimmune prone mouse model we introduced a Fas mutation in the CD2CREMαtg mice (FVB/Fas^–/–CD2CREMαtg). Overexpression of CREMα strongly accelerated the lymphadenopathy and splenomegaly in the FVB/Fas^–/– mice. This was accompanied by a massive expansion of double‐negative (DN) T cells, enhanced numbers of interferon (IFN)‐γ‐producing T cells and reduced percentages of T_regs. Treatment of FVB/Fas^–/–CD2CREMαtg mice with IL‐2 restored the percentage of T_regs and reversed increased IFN‐γ production, but did not affect the number of DNTs. Our data indicate that CREMα contributes to the failure of tolerance in SLE by favouring effector T cells and decreasing regulatory T cells, partially mediated by repression of IL‐2 in vivo .     

Regulation of neuroinflammation by B cells and plasma cells

The remarkable success of anti‐CD20 B cell depletion therapies in reducing the burden of multiple sclerosis (MS) disease has prompted significant interest in how B cells contribute to neuroinflammation. Most focus has been on identifying pathogenic CD20⁺ B cells. However, an increasing number of studies have also identified regulatory functions of B lineage cells, particularly the production of IL‐10, as being associated with disease remission in anti‐CD20–treated MS patients. Moreover, IL‐10–producing B cells have been linked to the attenuation of inflammation in experimental autoimmune encephalomyelitis (EAE), the animal model of MS. In addition to IL‐10–producing B cells, antibody‐producing plasma cells (PCs) have also been implicated in suppressing neuroinflammation. This review will examine regulatory roles for B cells and PCs in MS and EAE. In addition, we speculate on the involvement of regulatory PCs and the cytokine BAFF in the context of anti‐CD20 treatment. Lastly, we explore how the microbiota could influence anti‐inflammatory B cell behavior. A better understanding of the contributions of different B cell subsets to the regulation of neuroinflammation, and factors that impact the development, maintenance, and migration of such subsets, will be important for rationalizing next‐generation B cell–directed therapies for the treatment of MS.     

Novel therapeutic compound tuftsin–phosphorylcholine attenuates collagen‐induced arthritis

Treatment with helminthes and helminthes ova improved the clinical symptoms of several autoimmune diseases in patients and in animal models. Phosphorylcholine (PC) proved to be the immunomodulatory molecule. We aimed to decipher the tolerogenic potential of tuftsin–PC (TPC), a novel helminth‐based compound in collagen‐induced arthritis (CIA) a mouse model of rheumatoid arthritis (RA). CIA DBA/1 mice were treated with TPC subcutaneously (5 µg/0.1 ml) or orally (250 µg/0.1 ml), starting prior to disease induction. The control groups were treated with PBS. Collagen antibodies were tested by enzyme‐linked immunosorbent assay (ELISA), cytokine protein levels by ELISA kits and regulatory T (T_reg) and regulatory B (B_reg) cell phenotypes by fluorescence‐activated cell sorter (FACS). TPC‐treated mice had a significantly lower arthritis score of 1.5 in comparison with control mice 11.8 (P < 0.0001) in both subcutaneous and orally treated groups at day 31. Moreover, histology analysis demonstrated highly inflamed joints in control mice, whereas TPC‐treated mice maintained normal joint structure. Furthermore, TPC decreased the titres of circulating collagen II antibodies in mice sera (P < 0.0001), enhanced expression of IL‐10 (P < 0.0001) and inhibited production of tumour necrosis factor (TNF)‐α, interleukin (IL)?17 and IL‐1β (P < 0.0001). TPC significantly expanded the CD4⁺CD25⁺ forkhead box protein 3 (FoxP3⁺) T_reg cells and CD19⁺IL‐10⁺CD5^highCD1d^highT cell immunoglobulin mucin‐1 (TIM‐1⁺) B_reg cell phenotypes (P < 0.0001) in treated mice. Our data indicate that treatment with TPC attenuates CIA in mice demonstrated by low arthritic score and normal joints histology. TPC treatment reduced proinflammatory cytokines and increased anti‐inflammatory cytokine expression, as well as expansion of T_reg and B_reg cells. Our results may lead to a new approach for a natural therapy for early rheumatoid arthritis onset.     

Impaired B‐cell tolerance checkpoints promote the development of autoimmune diseases and pathogenic autoantibodies

A role for B cells in autoimmune diseases is now clearly established both in mouse models and humans by successful treatment of multiple sclerosis and rheumatoid arthritis with anti‐CD20 monoclonal antibodies that eliminate B cells. However, the underlying mechanisms by which B cells promote the development of autoimmune diseases remain poorly understood. Here, we review evidence that patients with autoimmune disease suffer from defects in early B‐cell tolerance checkpoints and therefore fail to counterselect developing autoreactive B cells. These B‐cell tolerance defects are primary to autoimmune diseases and may result from altered B‐cell receptor signaling and dysregulated T‐cell/regulatory T‐cell compartment. As a consequence, large numbers of autoreactive naive B cells accumulate in the blood of patients with autoimmune diseases and may promote autoimmunity through the presentation of self‐antigen to T cells. In addition, new evidence suggests that this reservoir of autoreactive naive B cells contains clones that may develop into CD27^?CD21^?/lo B cells associated with increased disease severity and plasma cells secreting potentially pathogenic autoantibodies after the acquisition of somatic hypermutations that improve affinity for self‐antigens.     

Anti‐CD25 monoclonal antibody Fc variants differentially impact regulatory T cells and immune homeostasis

Interleukin‐2 (IL‐2) is a critical regulator of immune homeostasis through its non‐redundant role in regulatory T (Treg) cell biology. There is major interest in therapeutic modulation of the IL‐2 pathway to promote immune activation in the context of tumour immunotherapy or to enhance immune suppression in the context of transplantation, autoimmunity and inflammatory diseases. Antibody‐mediated targeting of the high‐affinity IL‐2 receptor α chain (IL‐2Rα or CD25) offers a direct mechanism to target IL‐2 biology and is being actively explored in the clinic. In mouse models, the rat anti‐mouse CD25 clone PC61 has been used extensively to investigate the biology of IL‐2 and Treg cells; however, there has been controversy and conflicting data on the exact in vivo mechanistic function of PC61. Engineering antibodies to alter Fc/Fc receptor interactions can significantly alter their in vivo function. In this study, we re‐engineered the heavy chain constant region of an anti‐CD25 monoclonal antibody to generate variants with highly divergent Fc effector function. Using these anti‐CD25 Fc variants in multiple mouse models, we investigated the in vivo impact of CD25 blockade versus depletion of CD25⁺ Treg cells on immune homeostasis. We report that immune homeostasis can be maintained during CD25 blockade but aberrant T‐cell activation prevails when CD25⁺ Treg cells are actively depleted. These results clarify the impact of PC61 on Treg cell biology and reveal an important distinction between CD25 blockade and depletion of CD25⁺ Treg cells. These findings should inform therapeutic manipulation of the IL‐2 pathway by targeting the high‐affinity IL‐2R.     

1,25‐dihydroxyvitamin D3‐induced dendritic cells suppress experimental autoimmune encephalomyelitis by increasing proportions of the regulatory lymphocytes and reducing T helper type 1 and type 17 cells

Dendritic cells (DCs), a bridge for innate and adaptive immune responses, play a key role in the development of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), an animal model for MS. Administration of tolerogenic DCs has been used as an immunotherapy in autoimmune diseases. Deficiency of vitamin D is an environmental risk factor of MS. In this study, we induced tolerogenic DCs by 1,25‐dihydroxyvitamin D₃ and transferred the tolerogenic DCs (VD₃‐DCs) into EAE mice by adoptive transfer. We found that VD₃‐DCs inhibited the infiltrations of T helper type 1 (Th1) and Th17 cells into spinal cord and increased the proportions of regulatory T cells (CD4⁺ CD25⁺ Foxp3⁺), CD4⁺ IL‐10⁺ T cells and regulatory B cells (CD19⁺ CD5⁺ CD1d⁺) in peripheral immune organs, which resulted in attenuated EAE. However, the proportions of T helper type 1 (Th1) and Th17 cells in spleen and lymph nodes and the levels of pro‐inflammatory cytokines and IgG in serum also increased after transfer of VD₃‐DCs. We conclude that transfer of VD₃‐DCs suppressed EAE by increasing proportions of regulatory T cells, CD4⁺ IL‐10⁺ T cells and regulatory B cells in spleen and reducing infiltration of Th1 and Th17 cells into spinal cord, which suggests a possible immunotherapy method using VD₃‐DCs in MS.     

Spontaneous downregulation of antibody/autoantibody synthesis in susceptible mice upon chronic exposure to mercuric chloride is not owing to a general immunosuppression

Administration of mercuric chloride into susceptible rats and mice induces a systemic autoimmune disease, which is characterized by a T‐cell‐dependent polyclonal B‐cell activation, an increase in serum levels of immunoglobulin (Ig)G1 and IgE, production of antibodies of different specificities and development of renal IgG deposits. A peculiar feature of mercury‐induced autoimmunity is that the polyclonal B‐cell activation spontaneously disappears in spite of continuous injection of mercury. The exact mechanism(s) for autoregulation of mercury‐induced autoimmunity is not well understood. In the present study, we analysed the regulation of mercury‐induced immune/autoimmune responses in mice and tested whether spontaneous downregulation of these responses is owing to a general immunosuppression. Mercury‐susceptible [SJL (H‐2^s)] and ‐resistant [DBA/2 (H‐2^d)] mice were injected with mercury for 4, 10, 15 and 17 weeks. Immune/autoimmune responses were monitored in these mice. Thereafter, mercury‐injected mice for 17 weeks were further immunized with horse red blood cells (HRBC) to study whether the subsequent humoral immune response to a foreign antigen is suppressed. We found that except for IgG1 anti‐nucleolar antibody production and renal IgG1 deposition, other characteristics of mercury‐induced autoimmunity were downregulated in SJL (H‐2^s) mice after chronic treatment with mercury. However, these mice did not show any reduction in the number of splenic antibody‐secreting cells and/or in serum titres of specific IgM, IgG1 and IgG2a anti‐HRBC antibodies in response to HRBC as compared with naïve mice. Similarly, in mercury‐resistant DBA/2 (H‐2^d) mice, chronic treatment with mercury did not either suppress specific antibody responses against HRBC. Our findings show that the autoregulation of mercury‐induced immune/autoimmune responses observed after chronic treatment with mercury is not owing to a general immunosuppression.     

Interleukin (IL)‐39 [IL‐23p19/Epstein–Barr virus‐induced 3 (Ebi3)] induces differentiation/expansion of neutrophils in lupus‐prone mice

Interleukin (IL)‐12 family cytokines play critical roles in autoimmune diseases. Our previous study has shown that IL‐23p19 and Epstein–Barr virus‐induced 3 (Ebi3) form a new IL‐12 family heterodimer, IL‐23p19/Ebi3, termed IL‐39, and knock‐down of p19 or Ebi3 reduced diseases by transferred GL7⁺ B cells in lupus‐prone mice. In the present study, we explore further the possible effect of IL‐39 on murine lupus. We found that IL‐39 in vitro and in vivo induces differentiation and/or expansion of neutrophils. GL7⁺ B cells up‐regulated neutrophils by secreting IL‐39, whereas IL‐39‐deficient GL7⁺ B cells lost the capacity to up‐regulate neutrophils in lupus‐prone mice and homozygous CD19^cre (CD19‐deficient) mice. Finally, we found that IL‐39‐induced neutrophils had a positive feedback on IL‐39 expression in activated B cells by secreting B cell activation factor (BAFF). Taken together, our results suggest that IL‐39 induces differentiation and/or expansion of neutrophils in lupus‐prone mice.