Anti‐annexin 1 antibodies: A new diagnostic marker in the serum of patients with discoid lupus erythematosus

Abstract: Annexin 1 is an anti‐inflammatory molecule and has also been described to be a common target of autoantibodies. In this study, we determined whether antibodies against annexin 1 can be detected in sera of patients with cutaneous lupus erythematosus (CLE). Levels of anti‐annexin 1 antibodies were evaluated by a new established enzyme‐linked immunosorbent assay and found to be significantly higher in sera of patients with CLE when compared to normal healthy donors (NHD). Moreover, the percentage of sera positively tested for anti‐annexin 1 antibodies was elevated in patients with CLE when compared to NHD. In particular, the percentage of positive sera for anti‐annexin 1 antibodies was significantly higher in patients with discoid lupus erythematosus (DLE); however, disease activity did not correlate with the antibody levels. The results of this study indicate that anti‐annexin 1 antibodies in sera of patients with DLE might be a valuable aid in the diagnosis of this subtype.     

Upregulation of epidermal surface molecule expression in primary and ultraviolet-induced lesions of lupus erythematosus tumidus

BACKGROUND: Lupus erythematosus tumidus (LET), a photosensitive skin disorder with characteristic clinical and histological features, has not been generally accepted as a subset of cutaneous lupus erythematosus (CLE). OBJECTIVES: To analyse the expression of epidermal surface molecules in skin biopsy specimens from patients with LET and to relate the results to other variants of CLE, such as discoid lupus erythematosus (DLE) and subacute CLE (SCLE). METHODS: In total, 45 patients with different subtypes of CLE were included in the study, and cryostat sections from primary and ultraviolet (UV) A- and UVB-induced skin lesions were investigated using immunohistochemical methods. RESULTS: In contrast to healthy controls, skin lesions of LET showed upregulation of intercellular adhesion molecule-1 (ICAM-1) and histocompatibility class II molecules (HLA-DR), with an expression pattern resembling that seen in DLE and SCLE. Furthermore, staining with a monoclonal antibody against 27E10, a distinct marker for cell activation and differentiation, revealed intense focal or band-like labelling of all epidermal layers independent of the type of lesion. CONCLUSIONS: Expression of epidermal surface molecules such as ICAM-1, HLA-DR and 27E10 is equally upregulated in primary and UV-induced lesions of patients with LET, DLE and SCLE. These results support our recent clinical findings that LET represents a distinct subset of CLE with a similar immunopathomechanism rather than a different disease.     

Ultraviolet light protection by a sunscreen prevents interferon‐driven skin inflammation in cutaneous lupus erythematosus

Irradiation with ultraviolet (UV) light is an important exacerbating factor in cutaneous lupus erythematosus (CLE) and induces various effects in the skin of patients with the disease, such as cell death and inflammation. Recently, we demonstrated the ability of a broad‐spectrum sunscreen to prevent UV‐induced damage both in patients with CLE and healthy controls (HCs). The aim of this study was to evaluate whether the UV‐dependent activation of interferon (IFN)‐driven inflammation in CLE can also be prevented by application of the sunscreen. In 20 patients with different subtypes of CLE and 10 HCs, defined areas on the upper back were treated with a broad‐spectrum liposomal sunscreen 20 min prior to a combined standardized UVA/UVB irradiation. Immunohistological analyses using antibodies directed against MxA, CD11c, CD123 and CD68 were performed from skin biopsies taken from areas before UV irradiation as well as from sunscreen‐treated and sunscreen‐untreated areas 24 and 72 h after UV irradiation. The expression of MxA was completely prevented by the sunscreen applied prior to UV irradiation in CLE patients and HCs. Additionally, sunscreen protection significantly diminished the number of the CD11c‐ and CD123‐positive dendritic cells, which are suggested to be a major source of type I/III IFNs, in UV‐irradiated skin of patients with CLE. Moreover, the application of the sunscreen prevented the increase in CD68‐positive macrophages in both groups 72 h after UV irradiation. The data of this study demonstrate that UV protection reduces lesional tissue damage and inhibits the typical IFN‐driven inflammatory response in CLE.     

以眼睑和头皮红斑为表现的早期皮肤型红斑狼疮一例

患者,女,29岁。左眼睑红斑3个月,发现头皮脱发区1天。头皮组织病理：表皮轻度角化过度,基底细胞灶性液化变性,真皮浅层血管周围少许单一核细胞浸润,真皮内毛囊数量明显减少,真皮深层毛囊及附属器周围较多淋巴细胞浸润。免疫荧光：表皮基底膜C3、IgM、IgA弱阳性带状沉积,IgG阴性。阿新蓝染色:胶原间黏蛋白沉积。诊断为皮肤红斑狼疮。给予硫酸羟氯喹片口服、他克莫司软膏和卤米松软膏外用治疗,红斑消退。     

Interleukin-6 expression in the skin of patients with lupus erythematosus

Abstract It has been proposed that interleukin-6 may play a role in the pathogenesis of autoimmune diseases like lupus erythematosus. We have therefore investigated the immunoreactivity of IL-6 in 32 skin biopsies of 23 patients suffering from chronic discoid lupus erythematosus (n=16), subacute cutaneous lupus erythematosus (n=5) and systemic lupus erythematosus (n = 5) as well as in uninvolved skin (n = 6) and in normal skin from healthy volunteers (n = 3). Increased immunohistochemical staining was detectable in 14 of 26 biopsies from lesional skin. The remaining biopsies from lesional, non-lesional and normal skin displayed only minimal or no reactivity, but 8 out of 12 lupus erythematosus patients had been pretreated with local or systemic antiinflammatory drugs. Irrespective of the LE subtype, immunolabelling was generally most intense in the basal layer of the epidermis, with additional intense suprabasal staining in sections from 2 of 5 SLE patients. Preferential production of IL-6 in the lower parts of the epidermis was confirmed by RNA in situ hybridization. No correlation was found between the deposition of immuno-globulins and complement at the dermo-epidermal junction and IL-6 expression in keratinocytcs. These data suggest that IL-6 may be involved in LE although its exact role in the pathogenesis of the disease needs to be further elucidated.     

Increased expression of guanylate binding protein-1 in lesional skin of patients with cutaneous lupus erythematosus

The large GTPase human guanylate binding protein-1 (GBP-1) is a key mediator of angiostatic effects of inflammation and is induced by interferon (IFN)-α and IFN-γ in endothelial cells (ECs). The aim of this study was to investigate whether GBP-1 is a marker of skin lesions in patients with cutaneous lupus erythematosus (CLE). Western blotting revealed that GBP-1 was in vitro induced by IFN-α and -γ in primary keratinocytes obtained from healthy controls. Moreover, we found that this protein was expressed by keratinocytes and ECs in primary and ultraviolet (UV)-induced skin lesions from patients with various subtypes of CLE, when compared to non-lesional skin. No GBP-1 expression was noted in skin biopsy specimens 24 or 72 h after UV irradiation prior to lesion formation in patients with CLE or in healthy control specimens with or without UV irradiation. Initial findings suggest that GBP-1 is not expressed in other skin diseases with different inflammatory aetiology, such as atopic dermatitis. We conclude that GBP-1 expression is closely associated with skin lesions in patients with CLE, suggesting a contribution of GBP-1 in the pathogenesis of this disease.     

Relevant new insights into the effects of photoprotection in cutaneous lupus erythematosus

Cutaneous lupus erythematosus (CLE) denotes a heterogeneous spectrum of autoimmune diseases that primarily affect the skin. Ultraviolet irradiation (UV) is one of the most important environmental factors that can trigger skin lesions in CLE or even induce systemic organ manifestation. It has been shown that broad‐spectrum sunscreen with high sun protection factors can effectively prevent UV‐induced skin lesions in patients with different subtypes of CLE. In a recent issue of Experimental Dermatology, Zahn and colleagues demonstrate that potent photoprotection blocks disease‐specific skin lesions in CLE patients by reducing lesional tissue damage and interferon‐driven inflammation.     

Measuring the activity of the disease in patients with cutaneous lupus erythematosus

The Systemic Lupus Activity Measure (SLAM) is a system proposed by rheumatologists to measure disease activity in their patients with systemic lupus erythematosus (LE). It involves scoring a group of clinical symptoms and laboratory findings, the maximum possible score being 84. In systemic LE, the mid-point is between 9 and 12. We applied SLAM to 176 patients with cutaneous LE. Ninety-seven had localized discoid LE (L-DLE), 59 had disseminated discoid LE (D-DLE) and 20 had subacute cutaneous LE (SCLE). Eighty-five patients had low activity disease (0-4 points), 72 mildly active disease (5-9 points), 15 moderately active disease (10-14 points) and only four had very active disease (>/= 15 points). The most frequent lesions in patients who scored more than 10 points were photosensitivity, cicatricial alopecia, Raynaud's phenomenon and oral ulcers. Fifty patients were followed up for more than 5 years (mean follow-up 9 years). Nine of these had an increased SLAM score. Seven had L-DLE, one D-DLE and one SCLE. Seven of the 50 patients had photosensitivity, five cicatricial alopecia, five non-cicatricial alopecia, two Raynaud's phenomenon and two oral ulcers. Three patients who started with L-DLE evolved to D-DLE. The SLAM system is useful in the monitoring of disease activity in patients with cutaneous LE. Over time, even L-DLE patients may develop active disease. Photosensitivity, alopecia, oral ulcers and Raynaud's phenomenon seem to herald a worse prognosis.     

Investigating the presence of neutrophil extracellular traps in cutaneous lesions of different subtypes of lupus erythematosus

Neutrophil extracellular traps (NETs) are implicated in the pathogenesis of systemic lupus erythematosus (SLE). However, little is known about the implication of NETs in cutaneous lupus. In this case series of 30 patients, we compared the amounts of neutrophils producing NETs in cutaneous lesions of different subtypes of lupus (5 with discoid lupus or DLE, 5 with subacute cutaneous lupus or SCLE, 11 with acute cutaneous SLE, 7 with lupus panniculitis and 2 with chilblains). Immunofluorescence was performed on formalin‐fixed paraffin‐embedded skin biopsies using antibodies against neutrophilic granules (elastase, myeloperoxidase, PR‐3 proteins and citrullinated histone 3). Dihydroethidium staining was performed to detect reactive oxygen species (ROS), known inducers of NETs. NETs were detected in the different subtypes of cutaneous lupus as well as in cutaneous lesions of SLE. The amounts of neutrophils producing NETs were significantly higher in lupus panniculitis (49%), acute cutaneous SLE (41%) and DLE (32%), in comparison with SCLE (5%) and chilblains (0%). This suggests that NETs might be associated with more tissue damage and scarring. ROS were observed in the different cutaneous subtypes of lupus independent of NETs.     

The expression pattern of interferon-inducible proteins reflects the characteristic histological distribution of infiltrating immune cells in different cutaneous lupus erythematosus subsets

BACKGROUND: Plasmacytoid dendritic cells and type I interferons (IFNs) are supposed to play a central proinflammatory role in the pathogenesis of cutaneous lupus erythematosus (LE). The IFN-inducible chemokines CXCL9 and CXCL10 are involved in recruiting CXCR3+ effector lymphocytes from the peripheral blood into skin lesions of LE. We hypothesized that the expression pattern of IFN-inducible proteins reflects the characteristic distribution of the inflammatory infiltrate in different subsets of cutaneous LE. OBJECTIVES: To test this hypothesis in patients with LE. METHODS: Lesional skin biopsies taken from patients with different subsets of LE [chronic discoid LE (CDLE), n = 12; subacute cutaneous LE (SCLE), n = 5; LE tumidus (LET), n = 4; LE profundus (LEP), n = 6] were investigated by immunohistochemistry using monoclonal antibodies to the lymphocyte surface markers CD3, CD4, CD8, CD20 and CD68, the cytotoxic proteins Tia1 and granzyme B, the chemokine receptor CXCR3, the specifically type I IFN-inducible protein myxovirus protein A (MxA) and the chemokines CXCL9 and CXCL10. RESULTS: The expression pattern of MxA followed the distribution of the inflammatory infiltrate typically seen in the investigated cutaneous LE subsets. In CDLE and SCLE, expression was focused in the epidermis and upper dermis, while in LET a perivascular and in LEP a subcutaneous pattern was found. Similar findings were obtained for CXCL9 and CXCL10. CONCLUSIONS: Our results demonstrate a close morphological association between the expression pattern of IFN-inducible proteins and the distribution of CXCR3+ CD3+ lymphocytes in all investigated subsets of cutaneous LE. This supports the importance of an IFN-driven inflammation in this condition. Infiltrating lymphocytes carrying CXCL10 in their granules might amplify the lesional inflammation and be responsible for the chronic course of this disease.     

Elevated levels of serum soluble Fas are associated with organ and tissue damage in systemic lupus erythematosus among Chinese

The aims of the present study are to evaluate the difference of the levels of soluble Fas (sFas) antigen between patients with systemic lupus erythematosus (SLE) and healthy controls and to explore whether sFas has a role in either the disease activity or the organ damage in SLE. Serum levels of sFas were measured in 40 Chinese patients with SLE and 15 age-, gender-, and race-matched healthy controls using double antibody ELISA. SLEDAI scores for disease activity were determined. Data of organ and tissue damage was obtained from clinical records. Serum sFas levels were significantly increased in both more active (mean=8043.8 pg/ml, P<0.001) and less active SLE patients (mean=4820.2 pg/ml, P<0.001) comparing to the healthy controls (mean=3253.4 pg/ml). There was also a significant difference in serum sFas levels between the more active SLE patients and less active SLE patients (P=0.04). But, the levels of sFas didn’t correlate with SLEDAI. There was a significant difference in the serum sFas levels between patients with and without CNS disease (mean=9582.6, 6634.5 pg/ml; P=0.007). The same was true when patients with and without renal disease (mean=10972.7, 6520.1 pg/ml; P=0.019), as well as serositis (mean=10385.3, 6709.1 pg/ml; P=0.005) were analyzed. sFas is elevated in sera of SLE patients, especially in patients with active SLE. The elevated levels of sFas in the sera of patients with SLE may be closely associated with damage to the kidneys, central nervous system and serosa. Serum sFas may serve as a predictor of some organ and tissue damage in SLE.     

Up-regulation of cellular FLICE-inhibitory protein in peripheral blood B lymphocytes in patients with systemic lupus erythematosus is associated with clinical characteristics

Background  Systemic lupus erythematosus (SLE) is an autoimmune disease which is involved in T- and B-lymphocyte–mediated autoimmunity. Apoptosis contributes to the maintenance of lymphocytes homeostasis and the deletion of autoreactive cells in SLE. Although there is evidence that cellular FLICE-inhibitory protein (c-FLIP), an antiapoptosis protein, is increased in human lupus T cells to keep them from apoptosis, but the expression of apoptosis-regulatory protein c-FLIP in SLE B lymphocytes remains unknown.
Aims  To study the expression of c-FLIP in peripheral blood B lymphocytes in SLE patients and to investigate the relationship among the expression of c-FLIP in peripheral blood B lymphocytes in SLE patients, clinical manifestation and the levels of interleukin-4 (IL-4) and IL-10.
Methods  In this study, we detected the expression of c-FLIP in peripheral blood B lymphocytes in SLE patients by flow cytometry and the levels of IL-4 and IL-10 in SLE serum samples by enzyme-linked immunosorbent assay and analysed their relationship with clinical characteristics.
Results  We observed a significantly higher percentage of c-FLIP in peripheral B cells in SLE patients with active disease when compared to inactive ones and healthy controls. And the expression of c-FLIP in lupus peripheral B cells showed positive correlations with SLEDAI, erythrocyte sedimentation rate, C-reactive protein, antinucleosome antibody titre, IL-4, and IL-10, and negative correlation with white blood cell count. Patients with lupus nephritis had higher levels of c-FLIP in peripheral B cells than patients without lupus nephritis.
Conclusion  Our data show that overexpression of c-FLIP is relevant to the activity and severity of SLE. Its overexpression might play a role in preventing B cell from apoptosis in SLE. The cause of c-FLIP overexpression may be due to the increase of IL-4 and IL-10 levels in SLE patients.     

JAK inhibitor ruxolitinib inhibits the expression of cytokines characteristic of cutaneous lupus erythematosus

This study was stimulated by the clinical observation of a rapid response of a chilblain lupus patient to treatment with JAK1/2‐kinase inhibitor ruxolitinib. We investigated the in vivo expression of phospho‐JAK2 in CLE skin samples as well as the immunomodulatory in vitro effect of ruxolitinib in cultured immortalized keratinocytes and in a 3D human epidermis model (epiCS). Our results demonstrate that ruxolitinib significantly decreases the production of CLE‐typical cytokines (CXCL10, CXCL9, MxA) and might be a promising drug for future clinical studies in patients with CLE and related autoimmune skin diseases.     

SLE干扰素调节因子-1基因表达及泌乳素的研究

目的 探讨SLE患者中干扰素调节因子-1（IRF-1）基因的表达情况及泌乳素（PRL）对其干预作用。方法 电化学发光仪检测30例SLE患者与20例正常人对照组在清晨静息状态下的血清PRL水平,按密度梯度离心法分离、培养外周血PBMC,将样本分为SLE患者组、正常人对照组、正常PRL的SLE患者组 + 重组人泌乳素（rhPRL）和正常人对照组 + rhPRL等４组。应用RT-PCR结合凝胶图象扫描技术检测IRF-1基因的表达情况。结果 SLE患者组与正常人对照组IRF-1基因表达的相对值分别为0.89 ± 0.21和0.78 ± 0.18,两组比较,差异有统计学意义（P ＝ 0.026）,高PRL的SLE患者组为1.06 ± 0.26,显著高于正常PRL的SLE患者组的0.82 ± 0.21（P ＝ 0.005）,而正常PRL的SLE患者组与正常人对照组间差异无统计学意义（P ＝ 0.514）。经rhPRL刺激后,正常PRL的SLE患者IRF-1基因表达的相对值为0.99 ± 0.22,显著高于rhPRL刺激前（P ＝ 0.036）,而正常人对照组在刺激前后差异无统计学意义。结论SLE中存在IRF-1基因的表达异常,高PRL水平可能是导致IRF-1基因异常表达的原因之一。     

1 000例系统性红斑狼疮患者皮肤黏膜损害临床分析

目的:探讨系统性红斑狼疮(SLE)皮肤黏膜损害与系统表现和预后的相关性。方法:回顾性分析我院1996年1月~2005年4月诊治的SLE患者1 000例。结果:1 000例SLE患者病程中有皮肤黏膜损害者795例(79.5%),主要发生部位为面部(59.8%)和指(趾)部(32.8%);以皮肤黏膜损害为首发症状者418例(41.8%),有皮肤黏膜损害组与无皮肤黏膜损害组进行比较发现前者女性患者发生率较高,患者年龄轻,且关节症状、肌炎/肌痛和抗SM( )的发生率显著高于后者;而肾脏、浆膜受累和抗ds-DNA( )的发生率则明显低于后者,但两者治疗结果无显著性差异。结论:SLE患者皮肤黏膜损害的发生率高,在SLE的诊断中有很高的价值,但皮肤黏膜损害与SLE预后的关系不大。     

Copy number variations of the human histamine H4 receptor gene are associated with systemic lupus erythematosus

Background Systemic lupus erythematosus (SLE) is a complex genetic disease; the histamine H4 receptor (HRH4) has been shown to be related to different kinds of autoimmune disorders; and copy number variations (CNVs) have been found to be associated with various types of diseases. Objectives To explore a possible association between HRH4 (formerly H4R) CNVs and the risk of SLE. Methods Genomic DNA and RNA from 340 patients with SLE and 392 healthy controls were extracted, and CNVs and mRNA levels of HRH4 were examined. Results The expression of HRH4 mRNA was significantly increased in patients with SLE compared with controls. Amplification of HRH4 copy numbers significantly increased the risk of SLE [P < 0·001, odds ratio (OR) 2·26, 95% confidence interval (CI) 1·50–3·40]. HRH4 amplifications also positively correlated with the incidence of arthritis (P = 0·019, OR 1·96, 95% CI 1·11–3·47), and proteinuria (P < 0·001, OR 2·95, 95% CI 1·73–5·00) and antinuclear antibody abnormalities (P < 0·001, OR 2·97, 95% CI 1·66–5·33). Deletions of HRH4 copy numbers were protective against proteinuria (P = 0·03, OR 0·50, 95% CI 0·26–0·94). Conclusion CNVs of the HRH4 gene are associated with SLE.     

Lymphocytic infiltration of the skin is a photosensitive variant of lupus erythematosus: evidence by phototesting

BACKGROUND: Lymphocytic infiltration of the skin (LIS) is a disorder in which photosensitivity has been suspected but never proven. OBJECTIVES: To carry out a systematic photobiological investigation in patients with LIS and to compare the photobiological features of LIS with those of other photosensitive disorders. METHODS: We performed provocative phototesting with ultraviolet (UV) A and UVB in 10 patients with active LIS. RESULTS: In all patients, UVA and/or UVB elicited abnormal papular phototest reactions resembling lesions of LIS both clinically and histologically. Lesions typically developed 3--6 days (mean +/- SD 100.8 +/- 20.9 h) after the first UV exposure. CONCLUSIONS: This characteristic latency interval together with certain clinical features, i.e. onset in summer, predilection for the face and persistence of the lesions, indicate that LIS is a photosensitive disorder distinct from polymorphic light eruption but indistinct from lupus erythematosus (LE). Both our photobiological findings and the effective treatment with hydroxychloroquine in half of our patients strengthen the proposal that the two entities LIS and LE tumidus are identical. As diagnosis cannot be made by histological, immunofluorescence or laboratory criteria, provocative phototesting may be a diagnostic aid in this disorder.