Increased CCR4 expression on circulating CD4(+) T cells in ankylosing spondylitis, rheumatoid arthritis and systemic lupus erythematosus

Previous studies have suggested that CCR4 is particularly important in the selective recruitment of various subsets of leucocytes in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). In this study, we examined the percentage of CD4(+)/CCR4(+) T cells within circulating lymphocytes in active ankylosing spondylitis (AS), RA and SLE patients. The clinical significance of CCR4 expression as well as possible associations between the expression and serum levels of tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma and interleukin (IL)-10 were also examined. Our results showed that the percentage of CD4(+)/CCR4(+) T cells was significantly elevated in AS and RA patients as compared with normal controls. The percentage was also significantly higher in SLE patients who had received no treatment with glucocorticoids or cytotoxic drugs (untreated SLE) than that in controls. In addition, the percentage of CD4(+)/CCR4(+) T cells showed significant positive correlations with the Bath ankylosing spondylitis disease activity index (BASDAI) in AS and with the SLE disease activity index (SLEDAI) in untreated SLE. Of all the cytokines examined, the elevated serum IL-10 level was closely correlated with the percentage of CD4(+)/CCR4(+) T cells in AS, RA and untreated SLE. These results suggest that CCR4 may be crucial in the pathogenesis of AS, RA and SLE. The percentage of CD4(+)/CCR4(+) T cells can serve as a useful marker for the activity of AS and untreated SLE.     

Increased interleukin-4 production by NK T cells in systemic lupus erythematosus.

It has been reported that production of interleukin (IL)-4, a T helper (Th)-2-type cytokine, might play an important role in the pathogenesis of systemic lupus erythematosus (SLE). On the other hand, it is known that NK1.1(+) cells which belong to CD4, CD8 double-negative, or CD4(+) cells are associated with initial IL-4 production and Th2 differentiation in mice although human equivalent cells are unknown. In order to study the profile of IL-4-producing cells in SLE, cytoplasmic IL-4 and various surface antigens on peripheral mononuclear cells were analyzed. Peripheral mononuclear cells were stimulated for 5 h by phorbol ester and ionomycin in the presence of monensin, fixed, and permeabilized with paraformaldehyde and saponin solution. Then cytoplasmic IL-4 and various surface antigens were analyzed by flow cytometry. IL-4-producing cells in SLE were phenotypically the same as those which produce IL-4 normally and frequently bore activated T-cell (CD7, CD25, CD28, CD29) and NK-cell markers (CD56, CD57). Double-negative T cells and CD57(+) T cells were increased in number and were more frequently positive for cytoplasmic IL-4 in SLE compared with normal controls and various infectious diseases. It was suggested that T cells with NK cell markers, CD57(+) T cells, which are known to extrathymically differentiate, might be involved in the pathogenesis of SLE as a counterpart of mouse NK1.1(+) cells.     

Increased expression of soluble cytotoxic T-lymphocyte-associated antigen-4 molecule in patients with systemic lupus erythematosus

A soluble form of cytotoxic T-lymphocyte-associated antigen-4 (sCTLA-4) was recently found and shown to possess a downregulatory function as a membrane-bound CTLA-4 molecule. The purpose of the study was to investigate the expression of sCTLA-4 molecule in patients with systemic lupus erythematosus (SLE). One hundred patients with SLE and 40 age- and sex-matched healthy individuals were enrolled in the study. The results showed that patients with SLE have significantly higher levels of sCTLA-4 in sera than healthy controls (21.6 +/- 12.3 ng/ml versus 5.9 +/- 5.4 ng/ml, P < 0.001). Increased expression of sCTLA-4 mRNA in peripheral blood mononuclear cells (PBMCs) was also found in SLE patients. However, we could not find a statistically significant correlation between the serum levels of sCTLA-4 and lupus disease activities. The reported CTLA-4 gene polymorphism in promoter region at position -318 did not affect the levels of sCTLA-4. To the best of our knowledge, this is the first report showing that patients with SLE have increased sCTLA-4 expression. However, the mechanism and role of increased sCTLA-4 in the pathogenesis of SLE remains elucidated.     

C-type virus expression in systemic lupus erythematosus.

Kidneys from patients with lupus nephropathy, non-lupus immune-complex glomerulonephritis and other renal diseases were examined by indirect immunofluorescence for antigens related to a C-type virus from human cells (HEL-12 virus). All 11 specimens of lupus nephropathy contained HEL-12 virus antigens deposited in the same pattern as the immune complexes. The intensity of immunofluorescence with anti-HEL-12 virus serum correlated with the extent of immune-complex deposition. In contrast, nine renal lesions other than lupus nephropathy and seven normal tissues did not react with anti-HEL-12 virus serum. Antibody eluted from one kidney with lupus nephropathy reacted by indirect immunofluorescence with human and dog cells infected with HEL-12 virus but not with uninfected control cells. These findings demonstrate a specific association of lupus nephropathy with a C-type viral antigen that is deposited as antigen-antiviral antibody complex.     

Increased hydrazine excretion associated with systemic lupus erythematosus

Thirty patients with systemic lupus erythematosus (SLE) were given isoniazid (INH) and their acetylation phenotypes determined. Of 30 patients, 2 were slow acetylators (7%), 28 rapid acetylators. This was approximately equivalent to the ratio in the control group (12%) and to the prevalence in the general population (10%). Urinary excretion of the metabolite hydrazine was determined in 10 SLE patients and 9 controls. The significantly higher hydrazine excretion from SLE patients as compared with that from controls suggested an aberration of metabolism in the SLE patients.     

Increased proportion of CD56bright natural killer cells in active and inactive systemic lupus erythematosus

Natural killer (NK) cells belong to the innate immune system but can also affect adaptive immune reactions. This immune regulatory function is often ascribed to the CD56(bright) subpopulation of NK cells that is prevalent in secondary lymphoid tissues and has potent cytokine-producing ability. The NK cells have been described as affecting autoimmune disease and stimulating B-cell production of antibodies, but their role in systemic lupus erythematosus (SLE) pathology has not been extensively studied. We have studied NK cells in SLE, a B-cell-driven systemic autoimmune disease, and phenotypically characterized peripheral blood NK cells in comparison to NK cells from patients with immunoglobulin A nephritis, rheumatoid arthritis and healthy individuals. We have found an increased proportion of CD56(bright) NK cells in SLE, regardless of disease activity. We detected a somewhat increased expression of the activating receptor NKp46/CD335 on NK cells from SLE patients, although neither the percentage of NK cells of all lymphocytes nor the expression of other NK receptors analysed (LIR-1/CD85j, CD94, NKG2C/CD159c, NKG2D/CD314, NKp30/CD337, NKp44/CD336, CD69) differed between patient groups. We show that type I interferon, a proinflammatory cytokine known to be abundant in SLE, can cause increases of CD56(bright) NK cells in vitro. We confirmed that serum levels of interferon-alpha were increased in active, but not in inactive, disease in the SLE patient group. In conclusion, we found an increased proportion of CD56(bright) NK cells in the blood of SLE patients, although it remains to be examined whether and how this relates to the disease process.     

Patterns of systemic lupus erythematosus expression in Europe

ObjectivesTo analyse the differences in disease expression of European SLE patients based on gender, age at diagnosis, and ethnicity.MethodsA two-year, retrospective, multicentre, observational study was carried out in five countries (France, Germany, Italy, Spain and the UK). Patients' clinical manifestations including disease activity, organ involvement, organ damage and flares were analysed.ResultsThirty-one centres enrolled 412 consecutive eligible patients (90.5% of women), with active disease, stratified by disease severity (half severe and half non-severe).Baseline characteristics included; mean (SD) age: 43.3 (13.6) years, SLE duration: 10.7 (8.0) years and age at disease diagnosis: 32.6 (13.0) years old. The mean (SD) SELENA-SLEDAI and SLICC/ACR scores were: 8.1 (6.7) and 0.82 (1.36), respectively. Over half of patients experienced flares (54.9%). The average number of annual flares was 1.01 (0.71) flares/year.In males compared to females, the renal system was more frequently active (53.8% vs 30.0%, p = 0.002), the mean SLICC/ACR score was higher (1.15 vs 0.79, p = 0.039) and the pulmonary system was more likely to be damaged (12.8% vs 3.8%, p = 0.010). Furthermore, patients diagnosed at younger age displayed more renal system activity (young: 56.3% vs adult: 33.4% vs elder: 8.9%, p < 0.001) and renal damage (25.0% vs 6.9% vs 2.2%, p = 0.018) compared to the others. The annual number of flares (1.13 vs 1.05 vs 0.81 flares/year, p < 0.0001), including the occurrence of severe flares (0.58 vs 0.51 vs 0.20, p < 0.0001), was also higher in these patients.Conversely, greater organ damage was observed in patients diagnosed at an older age compared to the others. The mean SLICC/ACR score was higher (1.31 vs young: 0.88 and adult: 0.78, p < 0.001) in patients diagnosed in the older age groups. The pulmonary (13.3% vs younger: 0% vs adult: 3.7%, p = 0.030) and cardiovascular (17.8% vs younger: 0% vs adult: 2.9%, p < 0.001) systems were more frequently damaged in these patients.Black African descents showed greater disease activity compared to Caucasian patients. They flared more often (77.1% vs 48.6%, p = 0.001) and experienced a greater number of annual flares (1.57 vs 0.89 flares/year, p < 0.0001), mainly more severe flares (0.89 vs 0.38/year, p < 0.0001). They also were more likely to experience renal system damage.ConclusionThe study showed clearly two patient subsets. The disease was the most active in Black African descents, and this phenomenon has never been described before in continental Europe. The disease was also more active in patients diagnosed at a younger or adult. Greater disease damage was observed in males and in patients diagnosed at an older age.     

Levamisole in systemic lupus erythematosus.

Two patients with systemic lupus erythematosus complicated with lupus nephritis were treated with levamisole, an immunomodulator. Clinical features, including urinary protein excretion and creatine clearance, were restored in one patient, also immunoparameters such as ANA, Ig-bearing cell number and PHA-skin test. The other patient did not respond to levamisole but did respond to cyclophosphamide.     

CD4 antibody therapy in systemic lupus erythematosus

The emergence of monoclonal antibody technology has fostered new therapeutic strategies for people with autoimmune diseases. One of the most promising of these strategies involves the use of CD4 monoclonal antibodies, which are effective in animal models for systemic lupus erythematosus, diabetes mellitus, rheumatoid arthritis, myasthenia gravis, and multiple sclerosis. The appeal of CD4 antibodies is enhanced by several factors: (1) their effectiveness does not depend on depletion of target cells; (2) they may block the host immune response to therapy, and (3) they have been well-tolerated in preliminary human trials. The principal obstacle to the use of CD4 monoclonal antibodies stems from their adverse effects on normal immune function.     

系统性红斑狼疮患者脱氧核糖核酸酶1基因表达研究

目的　研究脱氧核糖核酸酶 1(deoxyribonuclease ,DNASE1)基因表达及 m RNA剪接形式与系统性红斑狼疮 (systemic lupus erythematosus,SL E)的关联性。　方法　以实时荧光定量聚合酶链反应 (real- time PCR)方法检测 DNASE1的 m RNA表达水平 ,以毛细管电泳技术分析 m RNA编码区替代剪接体 ,结合单核苷酸多态性 (single nucleotide polymorphisms,SNPs)单倍型了解基因结构对表达的影响。　结果　SL E患者 DNASE1基因表达水平显著高于正常对照 (P<0 .0 0 1) ,未发现 SL E疾病活动性指数积分与基因表达水平存在相关性 ,但性别分析显示女性患者 DNASE1基因表达水平高于男性患者 (P<0 .0 1)。 8名正常人与 18例患者的毛细管电泳结果显示 ,患者与正常人替代剪接谱系不同 ,且 380 bp处存在明显条带 ,具有不同单倍型的 SL E患者替代剪接谱系亦有差异。　结论　 SL E患者 DNASE1基因表达异常 ,并存在与正常人不同的 m RNA剪接体。 DNASE1基因与 SL E发病相关     

Natural autoantibodies in systemic lupus erythematosus.

We have tested the sera of 25 patients with systemic lupus erythematosus (SLE) for antibody activity against a panel of six antigens: DNA, TNP, actin, tubulin, myosin, albumin. Eluates from renal biopsy tissue were also tested. Sera from patients with lupus nephritis were found to contain high titres of IgA antibodies directed against the antigens of the panel, and marked IgG anti-DNA and anti-TNP antibody activity. The IgG anti-TNP antibodies isolated from SLE serum by affinity chromatography on a TNP-immunoadsorbent, were also found to possess anti-DNA activity. Kidney eluates obtained from biopsy specimens of SLE patients contained IgG antibodies strictly specific for DNA in three out of the nine patients tested, while three eluates from the remaining six patients reacted with DNA and TNP and three with DNA and all the other antigens of the panel. These results strongly suggest that in SLE sera there are at least three populations of circulating anti-DNA antibodies: those strictly specific for DNA, those recognizing DNA and TNP and those recognizing DNA and other macromolecules. Furthermore, because six out of nine of the eluates contained antibodies with an absolute or restricted specificity for DNA, this suggests that these antibodies are more often pathogenic than the polyspecific ones recognizing DNA and other macromolecules.     

Aortic aneurysm in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is frequently associated with cardiovascular manifestations, but rarely complicated with aortic disease. We report a 28-year-old female patient with a 14-year history of SLE and a 3-year history of hypertension. She had suffered from palpitation and chest tightness for 1 month before admission. Heart echo showed thoracic to low abdominal level with low flow. A computed tomography (CT) scan confirmed aneurysms of the descending thoracic and upper abdominal aorta, down to the renal level. Diagnosis of aortic aneurysm should be considered in patients with SLE, especially those who have a history of hypertension, prolonged steroid use, palpitation and chest pain. Current imaging modalities, such as cardiac echo, CT and magnetic resonance angiography may provide earlier detection of subclinical disease, which may aid in preventing these fatal complications. It is important to control hypertension aggressively in patients with SLE. In addition to decreasing steroid doses, early use of immunosuppressive agents and accurate noninvasive image modalities may allow us to prevent severe damage to the aorta and avoid the fatal complications.     

Neonatal lupus erythematosus in offspring of mothers with experimental systemic lupus erythematosus.

Neonatal lupus erythematosus (NLE) syndrome is a result of the transfer of autoantibodies produced by the mother, across the placenta, to the fetus. NLE is characterized by a transient dermatitis, a variety of systemic and hematological abnormalities, and isolated cases of congenital heart block. The latter has been reported to be due to the presence of autoantibodies specific to La (SS-B) and/or Ro (SS-A). As female mice with experimental SLE, induced by immunization with the monoclonal anti-DNA 16/6 Id, produce a variety of autoantibodies including anti-Ro and anti-La antibodies, we examined the relevance of NLE in the murine system. Offspring of SLE-afflicted BALB/c mothers possessed antibody titers to the 16/6 Id, ssDNA, and nuclear extract, which gradually declined until reduced to normal levels by day 60 after delivery. Antibody titers in the sera of the mothers remained elevated throughout this period. Electrocardiograms were recorded from groups of neonates from mothers with experimental SLE. The results indicated that a high percentage of the offspring had defects in their conduction system including first, second, and third degree heart block; significant bradycardia; and wide QRS complex. Normal patterns were observed in offspring of healthy mothers. Experiments done with mice that were exposed to SLE-related autoantibodies early in their development indicated that offspring to mothers with experimental SLE were neither protected nor more susceptible to disease induction by the 16/6 Id.     

Penicillin hypersensitivity in systemic lupus erythematosus.

The prevalence of positive immediate-type skin tests to penicillin antigens among patients with systemic lupus erythematosus (SLE) was compared with hospitalized controls without SLE and found not to differ significantly. On the other hand, we confirmed previous reports that the prevalence of histories of prior penicillin reaction was significantly higher in SLE patients than among controls. This is felt to be related to the primary disease process, and we conclude that patients with SLE do not have an increased risk of IgE-mediated allergic reactions to penicillin.     

Tissue c-myc protein expression and immune response in systemic lupus erythematosus.

Frozen sections of kidney tissue from 12 patients with systemic lupus erythematosus were examined for the expression of oncogene proteins. Polyclonal rabbit antibody to c-myc peptide or to whole 65,000 D c-myc protein, was used to identify c-myc protein within tissue sections. Patterns of nuclear staining in the Hep-2 cell line were similar to those seen with monospecific human SLE sera showing anti-Sm reactivity. In addition c-myc staining was not abolished by prior incubation of tissue sections with human serum containing anti-Sm antibodies. Five of 12 kidney biopsy tissues from patients with systemic lupus erythematosus (SLE) showed positive speckled c-myc protein staining within nuclei of monocyte/macrophage cells of glomerular tufts. Positive staining was in all instances completely abolished by prior absorption of anti-c-myc antibody with c-myc protein. No c-myc protein was identified within SLE immune complex deposits.     

HLA-G genotype and HLA-G expression in systemic lupus erythematosus: HLA-G as a putative susceptibility gene in systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune disease mainly mediated by the deposit of immune complexes and defects in T lymphocytes and antigen-presenting cells along with a high production of T-helper 2 cytokines. A tolerance-inducible function of nonclassical class Ib human leukocyte antigen (HLA)-G molecule in innate and adaptive cellular responses has been reported, suggesting a role in inflammatory diseases. A 14 bp sequence insertion/deletion polymorphism (rs16375) in the 3'-untranslated region of the HLA-G gene has been associated to the stability of HLA-G messenger RNA. The insertion of the 14 bp sequence seems to be associated with lower levels of soluble HLA-G (sHLA-G). The aim of this study was to evaluate the possible association of the presence of the 14 bp sequence (+14 bp) with SLE. We have HLA-G genotyped 200 SLE patients and 451 healthy control subjects (HS; Italian) and analyzed the plasma levels of sHLA-G and interleukin-10 (IL-10) in a subset of SLE patients and healthy subjects (Italian and Danish). A significant increase of the +14 bp HLA-G allele was detected in the Italian SLE patients compared with HS [P = 0.003, OR 1.44 (95% CI 1.13-1.82)]. A significant increased frequency of HLA-G +14/+14 bp and a decreased frequency of HLA-G -14/-14 bp were observed in SLE patients. There median concentration of sHLA-G was significantly lower in the plasma of SLE patients compared with that in the plasma of healthy controls (P < 0.0001). Furthermore, the results confirmed higher concentrations of IL-10-positive plasma in SLE patients. These results support a potential role for HLA-G in the susceptibility of SLE.