Treatment of Induced Murine SLE with a Peptide Based on the CDR3 of an Anti-DNA Antibody Reverses the Pattern of Pathogenic Cytokines

A peptide based on the complementarity determining region (CDR) 3 of a pathogenic anti-DNA monoclonal antibody that bears the 16/6 idiotype (Id) was shown previously to be a dominant T-cell epitope in experimental SLE, and to be capable of inhibiting SLE-associated responses. When injected, concomitant with active immunization with the pathogenic human anti-DNA, 16/6 Id + mAb, pCDR3 inhibited the proliferation of LN-derived T cells stimulated in vitro with the 16/6 Id mAb. The inhibition of the specific proliferative responses could be reversed by the addition of exogenous IL-2 to the cultures. Analysis of secreted cytokine profile in supernatants of these cultures demonstrated that pCDR3 treatment reduced significantly the levels of both IL-2 and IFN- &#110 that were elevated further in cells of the 16/6 Id-immunized mice. The CDR3-based peptide was shown here to immunomodulate in vivo experimental SLE, induced by the human anti-DNA 16/6 Id + antibody. The beneficial effects of pCDR3 on the clinical manifestations of SLE were associated with downregulation of the Th1-type (IL-2, IFN- &#110 ) and proinflammatory (TNF- &#102 ) cytokines, whereas the immunosuppressive cytokine TGF- &#103 was up regulated.     

The SLAM family member CD48 (Slamf2) protects lupus-prone mice from autoimmune nephritis

Polymorphisms in the SLAM family of leukocyte cell surface regulatory molecules have been associated with lupus-like phenotypes in both humans and mice. The murine Slamf gene cluster lies within the lupus-associated Sle1b region of mouse chromosome 1. Non-autoreactive C57BL/6 (B6) mice that have had this region replaced by syntenic segments from other mouse strains (i.e. 129, NZB and NZW) are B6 congenic strains that spontaneously produce non-nephritogenic lupus-like autoantibodies. We have recently reported that genetic ablation of the SLAM family member CD48 (Slamf2) drives full-blown autoimmune disease with severe proliferative glomerulonephritis (CD48GN) in B6 mice carrying 129 sequences of the Sle1b region (B6.129CD48^−/−). We also discovered that BALB/c mice with the same 129-derived CD48-null allele (BALB.129CD48^−/−) have neither nephritis nor anti-DNA autoantibodies, indicating that strain specific background genes modulate the effects of CD48 deficiency. Here we further examine this novel model of lupus nephritis in which CD48 deficiency transforms benign autoreactivity into fatal nephritis. CD48GN is characterized by glomerular hypertrophy with mesangial expansion, proliferation and leukocytic infiltration. Immune complexes deposit in mesangium and in sub-endothelial, sub-epithelial and intramembranous sites along the glomerular basement membrane. Afflicted mice have low-grade proteinuria, intermittent hematuria and their progressive renal injury manifests with elevated urine NGAL levels and with uremia. In contrast to the lupus-like B6.129CD48^−/− animals, neither BALB.129CD48^−/− mice nor B6 × BALB/c F1.129CD48^−/− progeny have autoimmune traits, indicating that B6-specific background genes modulate the effect of CD48 on lupus nephritis in a recessive manner.     

CRRT联合免疫吸附对重症系统性红斑狼疮患者疾病活动指数评分(SLEDAI)及预后转归影响的研究*

目的 观察CRRT联合免疫吸附对重症系统性红斑狼疮患者疾病活动指数评分(SLEDAI)及预后转归的影响。方法 将31例重症SLE患者分为对照组、IA组及CRRT+IA组。所有患者入院后均按指南要求给予糖皮质激素和免疫抑制剂药物治疗。IA组采用在此基础上给予DNA免疫吸附治疗。CRRT+IA组在IA组基础上联合CRRT(连续性静-静脉血液滤过模式CVVH)治疗。疗程为收住风湿免疫科(重症监护病房)后连续3日。分析三组患者治疗前后SLE疾病活动指数评分(SLEDAI)、多器官功能障碍综合征(MODS)发生率以及28天死亡率。结果 IA组SLE疾病活动指数评分(SLEDAI)及多器官功能障碍综合征(MODS)发生率下降显著(P＜0.05);CRRT+IA组较IA组SLE疾病活动指数评分(SLEDAI)下降显著(P＜0.05)。结论：对于重症系统性红斑狼疮患者,在传统的药物治疗基础上早期应用CRRT联合DNA免疫吸附这种多血液净化模式(集成血液净化模式)治疗,能显著改善患者临床症状,降低疾病严重程度,从而可能影响疾病的转归及预后。     

Plasmapheresis Modulates TH1/TH2 Imbalance in Patients with Systemic Lupus Erythematosus According to Measurement of Intracytoplasmic Cytokines

To examine the possible effect of plasmapheresis on the ratio of Th1/Th2 type cytokine-secreting cells we recruited eight patients with active systemic lupus erythematosus into the present study. They all failed to respond to conventional therapy. A sensitive multiparametric flow cytometric analysis was used for the detection of intracellular IL-4, IL-10 and IFN &#110 . Stimulated peripheral blood cells were analysed by this procedure. Plasmapheresis was performed every second day for three occasions, using a continuous flow type blood cell separator, and a total of 100 &#117 ml/body weight kg plasma was removed. Patients received 1 &#117 mg/kg/day methylprednisolone during this period. As a result of the procedure, the rate of IFN &#110 positive Th cells increased, while the rate of IL-4 and IL-10 expressing CD4 positive cells decreased. Together with these observations the concentration of anti-ds-DNA antibodies decreased after plasmapheresis. A decrease in disease activity index (SLE-DAI) indicated the clinical effectiveness of the therapy.     

Effect of Pro-inflammatory/Anti-inflammatory Agents on Cytokine Secretion by Peripheral Blood Mononuclear Cells in Rheumatoid Arthritis and Systemic Lupus Erythematosus

We studied a well-selected population of patients with active rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) without immunosuppressive therapy. Control and patient peripheral blood mononuclear cells (PBMC) were incubated with IL-1 &#103, IL-10, TGF- &#103 or LPS for 20 h and the in vitro basal and stimulated secretions of IL-6, TNF- &#102, IL-1 &#103 and IL-1ra were measured by ELISA. We found that in the SLE patients the basal secretion of IL-6 was significantly lower and that of IL-1ra significantly higher than in control subjects, while in the RA group the basal IL-1ra secretion was higher than in healthy subjects. SLE and RA PBMC responded to LPS and IL-1 &#103 reaching higher cytokine secretion values than controls. The in vitro response of SLE and RA PBMC to TGF &#103 was normal, while that to IL-10 was defective: IL-10 was able to stimulate the production of IL-6 and IL-1ra in PBMC from normal subjects, but it was unable to enhance IL-6 secretion in RA cells and it was also completely ineffective in inducing IL-1ra secretion in both SLE and RA PBMC. Our work add new data useful for the evaluation of IL-10 and IL-1ra as therapeutic agents in rheumatic diseases.     

Ly108 expression distinguishes subsets of invariant NKT cells that help autoantibody production and secrete IL-21 from those that secrete IL-17 in lupus prone NZB/W mice

Lupus is a systemic autoimmune disease characterized by anti-nuclear antibodies in humans and genetically susceptible NZB/W mice that can cause immune complex glomerulonephritis. T cells contribute to lupus pathogenesis by secreting pro-inflammatory cytokines such as IL-17, and by interacting with B cells and secreting helper factors such as IL-21 that promote production of IgG autoantibodies. In the current study, we determined whether purified NKT cells or far more numerous conventional non-NKT cells in the spleen of NZB/W female mice secrete IL-17 and/or IL-21 after TCR activation in vitro, and provide help for spontaneous IgG autoantibody production by purified splenic CD19⁺ B cells. Whereas invariant NKT cells secreted large amounts of IL-17 and IL-21, and helped B cells, non-NKT cells did not. The subset of IL-17 secreting NZB/W NKT cells expressed the Ly108^loCD4⁻NK1.1⁻ phenotype, whereas the IL-21 secreting subset expressed the Ly108^hiCD4⁺NK1.1⁻ phenotype and helped B cells secrete a variety of IgG anti-nuclear antibodies. α-galactocylceramide enhanced the helper activity of NZB/W and B6.Sle1b NKT cells for IgG autoantibody secretion by syngeneic B cells. In conclusion, different subsets of iNKT cells from mice with genetic susceptibility to lupus can contribute to pathogenesis by secreting pro-inflammatory cytokines and helping autoantibody production.