Active Systemic Lupus Erythematosus Group Research Articles

Leptin and adiponectin in patients with systemic lupus erythematosus: clinical and laboratory correlations

ObjectiveTo evaluate the serum levels of leptin and adiponectin in patients with systemic lupus erythematosus (SLE) and correlate their levels with disease activity, presence of autoantibodies and clinical manifestations. Methods52 women with SLE and 33 healthy women were evaluated. The patients were divided into two groups, the first with active SLE and the second with inactive SLE. Patients with SLEDAI ≥3 were considered active. Serum levels of leptin (ng/mL) and adiponectin (μg/mL) were measured by enzyme immunoassay. ResultsThere was a significant difference in leptin levels between SLE and controls (20.7±17.1 vs. 8.0±5.0ng/mL, p<0.001), but no significant difference in adiponectin levels (87 5±69.7 vs. 118.1±70.6pg/mL, p=0.053). No significant difference in levels of leptin and adiponectin was noted between inactive and active SLE groups. There was a significant association between low levels of leptin and positivity for anticardiolipin (aCL) (p=0.025) and lupus anticoagulant (LA) (p=0.003) and a significant association between high levels of leptin and the presence of renal disease (p<0.001). However, there was no association between adiponectin levels with autoantibodies and clinical features in SLE patients. ConclusionPatients with SLE had elevated leptin levels, with association with renal involvement. Leptin and adiponectin were not correlated with disease activity. Low levels of leptin have been associated with the presence of LA and aCL.

Validity and reliability of the Lupus QoL index in Turkish systemic lupus erythematosus patients.

Systemic lupus erythematosus (SLE) patients have seriously impaired quality of life (QoL). In addition to activity and damage indices used in the past, tools to evaluate QoL in SLE have been developed in recent years. In this study, we test the validity of the Turkish version of the Lupus-QoL (LupusQoL-TR) score, and investigate its association with clinical findings and activity indices. A total of 132 patients diagnosed with SLE according to ACR 1997 criteria were included. The clinical and demographic features, and biochemical data were retrieved from hospital records. SLE Disease Activity Index (SLEDAI) and damage score (SLICC-ACR) were determined at the time of administration of Lupus-QoL questionnaire. The Lupus-QoL includes 34 questions divided into eight domains. We reevaluated the LupusQoL-TR and pretested its understandability. SLE patients were concomitantly administered the LupusQoL-TR and generic SF-36. Internal consistency, test-retest reliability, convergent and discriminant validity were calculated. The mean age of our SLE patients was 37.9 ± 12.8 years. Internal consistency reliability ranged from 0.88 to 0.93, and test-retest reliability from 0.84 to 0.94. LupusQoL-related domains in SF-36 were correlated (from 0.66 to 0.74). Most LupusQoL-TR domains, except planning, were able to discriminate between active and inactive SLE groups. Scores in all domains of the LupusQoL-TR were found to be discriminative for patients with and without damage according to SLICC-ACR score. The LupusQoL-TR was found to be a valid patient-reported outcome measure method when evaluating QoL in Turkish SLE patients.

Investigating the role of angiogenesis in systemic lupus erythematosus

We aimed to elucidate the roles of six pro-angiogenic factors, namely, basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), placental growth factor (PlGF), hepatocyte growth factor (HGF), transforming growth factor-beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha), in the pathogenesis of systemic lupus erythematosus (SLE). A group of 75 patients with SLE and a control group of 40 healthy volunteers were recruited. Angiogenic factors were tested through enzyme-linked immunosorbent assay (ELISA) measurement. The angiogenic activities of the patients with SLE and the healthy controls were assessed and a correlation analysis of these angiogenic factors was conducted. A much higher level of angiogenic activity was shown in the serum of patients with SLE than that of the healthy controls, yet no statistically significant difference was found in the angiogenic activities of active SLE and inactive SLE. Serum VEGF level in the active SLE group was significantly higher than that in the control group and the inactive SLE group, and serum HGF level was strongly positively correlated with VEGF in all SLE groups; meanwhile, there was also a statistically significant positive correlation between TNF-alpha and VEGF in all SLE groups. There was a statistically significant positive correlation between serum VEGF level and bFGF level in the active SLE group. There was a slightly negative correlation between serum HGF level and TGF-beta level in the SLE group, but this negativity did not reach the significance level. Likewise, positive correlation was also shown in the serum VEGF level and PlGF level, yet not in bFGF with PlGF. Circulating serum angiogenic cytokines may be disease markers of SLE activity. Anti-angiogenic agents such as thalidomide and endogenous angiogenesis inhibitors such as endostatin are potentially effective and promising therapies in the treatment of SLE.

Leptina e adiponectina no lúpus eritematoso sistêmico: correlações clínicas e laboratoriais

ObjectiveTo evaluate the serum levels of leptin and adiponectin in patients with systemic lupus erythematosus (SLE) and correlate their levels with disease activity, presence of autoantibodies and clinical manifestations. Methods52 women with SLE and 33 healthy women were evaluated. The patients were divided into two groups, the first with active SLE and the second with inactive SLE. Patients with SLEDAI ≥3 were considered active. Serum levels of leptin (ng/ml) and adiponectin (μg/ml) were measured by enzyme immunoassay. ResultsThere was a significant difference in leptin levels between SLE and controls (20.7 ± 17.1 vs. 8.0 ± 5.0 ng/mL, P <0.001), but no significant difference in adiponectin levels (87.5 ± 69.7 vs. 118.1 ± 70.6 pg/ml, P = 0.053). No significant difference in levels of leptin and adiponectin was noted between inactive and active SLE groups. There was a significant association between low levels of leptin and positivity for anticardiolipin (aCL) (P = 0.025) and lupus anticoagulant (LA) (p = 0.003) and a significant association between high levels of leptin and the presence of renal disease (p <0.001). However, there was no association between adiponectin levels with autoantibodies and clinical features in SLE patients. ConclusionPatients with SLE had elevated leptin levels, with association with renal involvement. Leptin and adiponectin were not correlated with disease activity. Low levels of leptin have been associated with the presence of LA and aCL.

Expression of Ets-1 and FOXP3 mRNA in CD4+CD25+ T regulatory cells from patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease with complex genetic predisposing factors involved. Ets-1 transcription factor plays an important role in the suppressive activity of CD4(+)CD25(+) Treg cells and stable expression of FOXP3. To find its potential role in the pathogenesis of SLE, we investigate the mRNA expression of Ets-1 and FOXP3 mRNA in CD4(+)CD25(+) Treg cells from patients with SLE. Real-time transcription-polymerase chain reaction analysis was used to determine the expression of Ets-1 and FOXP3 mRNA in CD4(+)CD25(+) Treg cells from 36 patients with SLE and 18 sex-and-age-matched healthy controls. The Ets-1 mRNA expression level was decreased in patients with SLE [0.225 (0.135, 0.337)] than healthy controls [0.528 (0.303, 0.681)] (P < 0.001). The expression levels of FOXP3 mRNA were lower in SLE patients [0.608 (0.272, 1.164)] than healthy controls [0.919 (0.690, 1.223)], but the difference was not significant (P = 0.106). Significant reduction in Ets-1 and FOXP3 expression was also found in new-onset SLE subgroup when compared with healthy controls (P < 0.001). The level of Ets-1 and FOXP3 mRNA was not significantly different in hyperactive and lower active SLE group when compared with inactive SLE group, respectively (P > 0.05). There were no significant differences between SLE with lupus nephritis (LN) and SLE without LN either (P > 0.05). Associations of Ets-1 and FOXP3 mRNA expression levels with major clinical and laboratory parameters of SLE patients were also analyzed. However, no significant association was found. Significant positive correlation was found between Ets-1 and FOXP3 mRNA expression in CD4(+)CD25(+) Treg cells from SLE patients (r = 0.698, P < 0.001). Our results found that the expression levels of Ets-1 mRNA were decreased in SLE patients and Ets-1 expression was positively correlated with the expression of FOXP3. It indicated that Ets-1 may play an important role in the stable expression of FOXP3 in CD4(+)CD25(+) Treg cells.

Detection of active P-glycoprotein in systemic lupus erythematosus patients with poor disease control

Active P-glycoprotein (P-gp) molecules have been shown to transport steroids out of peripheral lymphocytes, resulting in poor responses to systemic steroid therapy in patients with systemic lupus erythematosus (SLE). This study was carried out to investigate the correlation between the expression or activity of P-gp in peripheral lymphocytes and disease control in SLE patients with a long history of systemic steroid treatment. A total of 60 SLE patients who had received systemic steroid treatment for longer than 6 months and 30 healthy subjects were monitored. SLE patients were subclassified into those with active and severely active forms of the disease according to their disease activity (estimated by SLEDAI-2000). The expression levels and activity of P-gp in peripheral blood lymphocytes were determined. Lymphocytes, obtained from three patients with severely active SLE, with high levels of P-gp expression were treated with cyclophosphamide, mycophenolic acid or emodin in vitro and Rh123-efflux activity was measured. P-gp expression in the peripheral lymphocytes of the SLE patients was significantly higher compared with that of the healthy controls, and a positive correlation between disease activity and P-gp expression levels was observed in these 60 patients. A significant increase in P-gp expression was observed in the severely active compared with the active SLE group. Treatment of lymphocytes with 100 μM cyclophosphamide or 100 μM emodin in vitro induced up to a 2-fold increase in the mean fluorescence intensity, as detected by the Rh123-efflux assay. In conclusion, the high expression levels of P-gp in the peripheral lymphocytes of SLE patients leads to poor disease control by systemic steroids. Emodin, an active ingredient derived from Chinese herbs, possesses a promising effect for overcoming P-gp-mediated steroid resistance by inhibiting the P-gp efflux function.

Pregnancy in women with systemic lupus erythematosus: a retrospective study of 111 pregnancies in Chinese women

Objective. The impact of pregnancy on lupus activity has been controversial especially in Chinese women. Research looking at predictive factors in this population are sparse. The aim of this study was therefore twofold: to determine the frequencies of abnormal pregnancy outcomes in a Chinese cohort and to identify clinical and laboratory factors predicting adverse fetal and maternal outcomes in Chinese women with systemic lupus erythematosus. Study design. Data of 111 pregnancies of 105 systemic lupus erythematosus (SLE) patients from January 1990 to December 2008 in Peking Union Medical College Hospital in Beijing were analyzed retrospectively. Univariate analysis using chi-square test and logistic regression was used to assess the predictive value of each variable on binary outcomes. Lupus activity was based on SLE Disease Activity Index (SLEDAI) criteria. Results. There were 23 elective, 2 spontaneous abortions, and 5 stillbirths, with 81 pregnancies resulting in live births including two multiple gestations. Three neonatal deaths were reported. Fetal loss rate including neonatal death was 11.1%. Fetal loss in active SLE group (17.0%) was significantly higher than those in inactive group (2.0%) (P = 0.047). The incidence of premature birth in active SLE group was 25/47 (53.2%), which is significantly higher than those in inactive group (3/34, 8.8%) (P < 0.001). Small for gestational age (SGA) was more common in active SLE group with incidence of 40.0% compared to 5.6% in inactive group (P < 0.001). Five fetal malformations were recorded (6.0%), including three fetal heart malformations (one complete heart block, one tetralogy of Fallot, and one atrial septal defect) and two multiple fetal malformations, which were significantly higher than general population. Among 25 pregnancies that were in active stage at conception, 14 (56%) deteriorated during pregnancy. Of the 68 pregnancies that were stable at conception, 26 (38%) flared during pregnancy or postpartum. Preeclampsia/eclampsia (OR = 14.83, 95% CI: 3.83–57.41) and thrombocytopenia (OR = 4.43, 95% CI: 1.12–17.60) were significant predictors of fetal loss; preeclampsia/eclampsia (OR = 8.04, 95% CI: 2.00–32.34) and active SLE (OR = 19.90, 95% CI: 2.38–166.27) were significantly associated with preterm birth; preeclampsia/eclampsia (OR = 8.92, 95% CI: 2.25–35.44) and thrombocytopenia (OR = 4.03, 95% CI: 1.24–17.25) were also significant predictors of maternal SLE flare-up. Conclusion. In general, lupus in pregnancy in the Chinese population is generally similar to other cohorts. Pregnancies can be successful in most women with SLE. However, an increase in SLE activity can occur in a significant number of patients, even those who are well controlled. Adverse fetal outcome including fetal loss, preterm birth, and SGA increases significantly with SLE flares during pregnancy with preeclampsia/eclampsia, thrombocytopenia, and active SLE serving independent predictors of adverse fetal and maternal outcome. Fetal echo should not just for heart block but for structural abnromalities as the structural malformation rate was significantly higher than general population, especially congenital heart disease.

Study of fas-mediated Foxp3+CD4+CD25+ Treg cell apoptosis in patients with systemic lupus erythematosus

To explore the role of fas apoptosis signal transduction pathway in the abnormal apoptosis of Foxp3+CD4+CD25+ Treg in patients with systemic lupus erythematosus (SLE). Twenty-five active SLE patients, 20 remission SLE patients and 25 controls were selected. The level of fas expression on peripheral blood Foxp3+CD4+CD25+ Treg surface was detected in SLE patients. And analyzed the expression rate of Foxp3 on CD4+CD25+ T cells was analyzed to explore the relationship between the expression rate and disease activity. (1) The expression rate of fas on Foxp3+CD4+CD25+ Treg was (23.72 ± 2.35)%, (14.0 ± 2.1)% in active and remission SLE groups respectively versus (10.1 ± 1.2)% in control group. The fas expression rate of active SLE group was significantly higher than those of remission SLE group(P < 0.01)and control group (P < 0.01). And the remission SLE and control groups were not statistically significant (P > 0.05). The expression rate of fas on the Foxp3+CD4+ CD25+ Treg was positively correlated with the SLEDAI (SLE disease activity index) score (r = 0.336, P < 0.05). (2) The expression rate of Foxp3 on CD4+CD25+ T cells was (2.83 ± 0.30)%, (5.38 ± 0.63)% in active and remission SLE groups respectively versus (8.12 ± 0.70)% in control group. The expression rate of Foxp3 was significantly lower in active SLE group than that in remission SLE group (P < 0.01)and control group (P < 0.01). And the Foxp3 expression rate of remission group was also lower than that of control group (P < 0.05). The expression rate of Foxp3 was negatively correlated with the SLEDAI score (r = -0.581, P < 0.01). (3) The expression rate of Foxp3 was negatively correlated with fas (r = -0.349, P < 0.01). The abnormal apoptosis of Foxp3+CD4+CD25+ Treg mediated by the fas apoptosis signal transduction pathway may be one of the pathogenic mechanisms of disease activity in SLE patients.

Regulatory T cells and their molecular markers in peripheral blood of the patients with systemic lupus erythematosus

CD4(+)CD25(+) regulatory T cells (Tregs) and the expression of their molecular markers (GITR, Foxp3) in peripheral blood of the patients with systemic lupus erythematosus (SLE) were investigated in order to reveal the pathogenesis of SLE on the cellular and molecular levels. The level of Tregs in peripheral blood was detected by flow cytometry. The expression levels of GITR and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR). The level of IL-6 in the plasma was measured by ELISA. Comparisons were made among 3 groups: the active SLE group, the inactive SLE group, and normal control group. The level of Tregs in the active SLE group and the inactive SLE group was significantly lower than in the normal control group (P<0.01). The level of Tregs in the active group was lower than in the inactive group with the difference being not significant (P>0.05). The level of Tregs in SLE patients was significantly negatively correlated with the disease active index in SLE (SLEDAI) (r=-0.81, P<0.01). The expression levels of GITR mRNA in PBMCs of the active SLE group and the inactive SLE group were significantly higher than in the normal control group (P<0.05), and those of Foxp3 mRNA in SLE patients of both active and inactive SLE groups were significantly lower than in the normal control group (P<0.05). There was no significant difference in the expression of GITR and Foxp3 mRNA between the active SLE group and inactive SLE group (P>0.05). The plasma levels of IL-6 in both the inactive SLE group and active SLE group were significantly higher than in the normal control group (P<0.01). The plasma level of IL-6 in the active SLE group was significantly increased as compared with that in the inactive SLE group (P<0.05), and the plasma level of IL-6 in SLE was significantly positively correlated with SLEDAI scores (r=0.58, P<0.01) and significantly negatively correlated with the ratio of CD4(+)CD25(+) cells/CD4(+) cells (r=-0.389, P<0.05). It was concluded that the levels of Tregs and Foxp3 mRNA in peripheral blood of SLE patients were decreased and the levels of GITR mRNA and plasma IL-6 were increased. The Tregs and their molecular markers GITR, Foxp3 as well as the plasma IL-6 might play an important role in the pathogenesis of SLE.

Oxidative stress and antioxidant parameters in a Turkish group of patients with active and inactive systemic lupus erythematosus

AbstractAim: Although the pathogenesis of systemic lupus erythematosus (SLE) is likely to be multifactorial, the inflammatory nature of SLE implies that a state of oxidative stress may exist in this disease, which may contribute to immune cell dysfunction, auto‐antigen production and auto‐antibody reactivity. This study was conducted to investigate the oxidant/antioxidant status of patients with SLE and to correlate this with disease activity.Methods: Sixteen clinically active, 12 clinically inactive SLE patients and 11 healthy controls were included into the study and antioxidant potential (AOP), catalase (CAT), glutathione peroxidase (GSH‐Px), superoxide dismutase (SOD), adenosine deaminase (ADA), malondialdehyde (MDA), and xanthine oxidase (XO) activity levels in erythrocytes and AOP, GSH‐Px, MDA, and XO activity levels in plasma were measured.Results: In erythrocytes; AOP was higher in SLE patients (both active and inactive), CAT activity was higher in inactive SLE patients, GSH‐Px, SOD and XO activity were lower in SLE patients (active and inactive) than the control group. In plasma; SLE patients (active and inactive) had higher AOP levels, activity of GSH‐Px was lower in SLE groups (active and inactive), activity of MDA was lower in active SLE patients, and activity of XO was higher in the active SLE patients than the control group.Conclusion: Antioxidant status was impaired in SLE patients but there was no significant difference between the active and inactive SLE groups, so oxidative stress may play a role in the pathogenesis of SLE but probably is not correlated with disease activity.

The expression of molecular chaperone HSP90 and IL-6 in patients with systemic lupus erythematosus

To explore the expression and clinical significance of molecular chaperone heat shock protein 90 (HSP90) in peripheral blood mononuclear cells (PBMC) and plasma level of interleukin-6 (IL-6) in patients with systemic lupus erythematosus (SLE), HSP90 was detected in PBMC by Western blot assay and the plasma level of IL-6 was measured by ELISA in 38 SLE patients and 20 normal controls. The correlation analysis was performed between the SLE disease activity index (SLE-DAI) and the expression of HSP90 and IL-6. The results showed that there was increased expression of HSP90 in the SLE patients. The active SLE group exhibited higher HSP90 levels (0.82+/-0.10) than the inactive SLE group (0.54+/-0.09) (P<0.01). The expression of HSP90 in normal control group (0.37+/-0.11) showed significant statistical difference as compared to both the inactive and active SLE groups (P<0.01, P<0.01, respectively). The plasma level of IL-6 exhibited a significant increase in both the inactive and active SLE groups (28.99+/-1.74 pg/mL, 44.58+/-9.15 pg/mL, respectively) compared with normal control group (P<0.01, P<0.01, respectively). The expression of HSP90 and IL-6 in SLE patients showed significant positive correlation with SLEDAI scoring (r=0.80, P<0.01: r= 0.74, P<0.01, respectively). In addition, there was a positive correlation between the level of IL-6 and HSP90 in SLE patients (r=0.86, P<0.01). The increased expression of molecular chaperone HSP90 and IL-6 may play an important role in the pathogenesis of SLE by regulating autoimmunity.

Messenger RNA expression of RANTES in the urinary sediment of patients with lupus nephritis

Lupus nephritis is characterized by intra-renal inflammation. Patients with systemic lupus erythematosus (SLE) showed abnormal T-cell expression of RANTES (regulated upon activation, normal T cell expressed) and its level in their serum. The authors studied the mRNA expression of RANTES in the urinary sediment of lupus patients. The authors studied 88 lupus patients, who were classified into active, remission and non-renal SLE groups according to the disease activity, 29 non-SLE and 10 healthy controls. Lupus activity was assessed by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Urinary mRNA expression of RANTES was studied by real-time quantitative polymerase chain reaction. The expression of RANTES in urinary sediment was significantly elevated in active group (P < 0.001). Expression level of RANTES correlated with the SLEDAI score (r = 0.57; P < 0.001) and renal score in SLEDAI (r = 0.60; P < 0.001). In addition, urinary expression of RANTES had significant correlation with degree of proteinuria, serum creatinine, albumin and estimated glomerular filtration rate. The authors conclude that the mRNA expression of RANTES was elevated in the urinary sediment of patients with active lupus nephritis. Measurement of urinary mRNA expression may be a novel non-invasive method for the assessment of lupus disease activity and the severity of renal involvement.

Expression of inducible co-stimulator in peripheral blood T lymphocytes in the patients with systemic lupus erythematosus.

The expression of inducible co-stimulator (ICOS) in peripheral blood T lymphocytes from the patients with systemic lupus erythematosus (SLE) and the role in the pathogenesis of SLE was investigated. By using two-color immunofluorescent staining and flow cytometric assay, the expression levels of ICOS in peripheal blood T lymphocytes from 33 patients with SLE and 16 healthy volunteers were detected. SLE diseases activity index (SLEDAI) of the patients with SLE was used to evaluate the disease activity. The correlation between the ICOS expression and SLEDAI was analyzed among the groups. The results showed that the expression levels of ICOS in T lymphocytes in active SLE group was markedly higher than those in the control and inactive SLE groups (both P< 0.01). There was no significant difference in the expression levels of ICOS between the inactive SLE and the control groups (P>0.05). In active SLE and inactive SLE groups, positive linear correlation was found between the levels of the ICOS expression in T lymphocytes and SLEDAI (r=0. 711, P=0.001; r=0.561, P=0.03). It was suggested that the expression of ICOS in peripheral blood T lymphocytes from the patients with active SLE was up-regulated and and ICOS might be related to the pathogenesis of SLE.

Defect in Regulation of Ca2+ Movement in Platelets from Patients with Systemic Lupus erythematosus

The differences in the intracellular Ca²⁺ responses to hormones in platelets from systemic lupus erythematosus (SLE) patients compared to normal humans have not been explored. This study examined the Ca²⁺ signaling and density of platelets in normal, inactive and active SLE patients. The platelet number per µl in inactive and normal groups did not differ, whereas the number in active SLE patients was smaller than the other two groups by 60%. The intracellular free Ca²⁺ levels ([Ca²⁺]_i) in response to stimulation of four endogenous Ca²⁺ mobilizing hormones, 100 µM arachidonic acid (AA), 10 µM ADP, 10 nM platelet activation factor (PAF) and 1 µM thrombin, were investigated using the Ca²⁺-sensitive fluorescent dye, fura-2. The AA-induced [Ca²⁺]_i rises in normal and inactive groups were similar. In contrast, the AA-induced [Ca²⁺]_i rises in the active SLE group were significantly smaller than in the normal and inactive groups. The defect in the AA-induced [Ca²⁺]_i rises in active SLE groups appears to be caused by defective Ca²⁺ influx and Ca²⁺ releasing pathways because the AA-induced responses were not altered by removal of extracellular Ca²⁺, whereas the AA-induced responses in normal and inactive SLE groups were reduced by removal of extracellular Ca²⁺, and the AA-induced Ca²⁺ release was smaller in the active SLE group. PAF, ADP and thrombin all induced [Ca²⁺]_i rises in the three groups, but no significant differences were found among the three groups. Together, the results indicate that cell density and Ca²⁺ signaling in platelets from active SLE patients are altered in response to particular stimulators. In these regards, platelets from inactive SLE patients appear to be similar to those from normal humans.

Interferon-inducible genes lymphocyte antigen 6 complex E and tetratricopeptide repeats 1 are correlated with clinical features of patients with systemic lupus erythematosus

To investigate the expression levels of lymphocyte antigen 6 complex, locus E (LY6E) and interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) genes in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), and to evaluate the relations between these gene expression levels and disease activity. The clinical data of 144 SLE patients, 27 non-SLE patients with rheumatic diseases, and 59 normal controls were collected. The SLE patients were further divided into 2 subgroups: active SLE group (n = 87) and non-active SLE group (n = 57) according to SLEDAI scores. Specimens of peripheral blood were drawn; total RNA was extracted and transcribed into cDNA. Sybr green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as-DeltaDeltaCT value) of LY6E and IFIT1 in patients with SLE and those in the controls. (1) The-DeltaDeltaCT value of LY6E expression level of the SLE patients was 5.4760 +/- 1.9806, significantly higher than those of the non-SLE patients (3.4323 +/- 1.7456) and normal controls (4.5198 +/- 1.6359, both P = 0.001). (2) The-DeltaDeltaCT value of LY6E and IFIT1 mRNA expression of the active SLE patients were 6.1960 +/- 1.7729 and 6.4997 +/- 2.6297 respectively, significantly higher than those observed in the inactive SLE patients (4.3770 +/- 1.7764 and 4.1327 +/- 2.6044 respectively, both P = 0.000). The-DeltaDeltaCT values of LY6E and IFIT1 mRNA of the SLE patients were correlated with the SLEDAI scores, and with the numbers of matched criteria used in the diagnosis of SLE (P < 0.001). As IFN-induced genes, elevated expression of LY6E mRNA is specific to diagnosis of SLE. The real time expression levels of LY6E and IFIT1 genes are associated with SLE disease activity. To inhibit the expression of LY6E and IFIT1 may become a novel therapeutic approach for SLE.

Roles of CCR2 and CXCR3 in the T cell-mediated response occurring during lupus flares.

Infiltrating lymphocytes have been demonstrated to play an important role in the tissue injury that occurs in systemic lupus erythematosus (SLE). Inflammatory chemokines control lymphocyte traffic through their interaction with T cell chemokine receptors. In this study we assessed the expression of chemokine receptors on T cell subsets of patients with active or inactive SLE. Forty-four SLE patients (40 women and 4 men) were included in the study. The patients were divided according to their SLE Disease Activity Index (SLEDAI), which resulted in a group of patients with inactive SLE (n = 27) and a group with active SLE (n = 17). The control group was composed of 22 healthy blood donors. A disease control group consisted of 18 patients infected with human immunodeficiency virus. Expression of chemokine receptors CCR1, CCR2, CCR5, CXCR3, CXCR4, and CX3CR1 was assessed on whole blood samples by immunofluorescence analysis. On T lymphocytes, significant differences between the SLE patients and controls were observed only in the expression of CCR2 and CXCR3. On monocytes, no significant differences in CCR2 expression were observed between the healthy controls and the SLE patients. The proportion of CD8+,CCR2+ T cells was significantly lower in the SLE patients compared with the controls (mean +/- SD 2.3 +/- 1.3% and 3.5 +/- 3.2% in the active and inactive SLE groups, respectively, versus 21 +/- 24% in controls; P < 0.0001 for both). The CD4+,CCR2+ subset was represented similarly among the controls and patients with inactive SLE (16.7 +/- 5.8% and 12.8 +/- 8.1%, respectively) but was depleted in patients with active SLE (7.1 +/- 4.4%; P < 0.0001 versus controls). The active SLE group expressed significantly lower circulating levels of CD4+,CCR2+ T cells than did the inactive disease group (P = 0.007). A negative correlation was found between the proportion of CD4+,CCR2+ T cells and the SLEDAI (r = -0.43, P = 0.005, by Spearman's correlation). Proportions of CD8+,CXCR3+ T cells were similar between the SLE groups and the control group (58 +/- 22.6% in active SLE, 47.1 +/- 20% in inactive SLE, and 59.4 +/- 17.3% in controls). The proportion of CXCR3-expressing CD4+ T cells was decreased in the active disease group (23.5 +/- 3.2% versus 39.9 +/- 12.5% in controls; P = 0.008) but not in the inactive disease group (34.8 +/- 9.5%). A trend toward a significant negative correlation was observed between the decreased proportion of CD4+,CXCR3+ T cells and the SLEDAI (P = 0.08). Following in vitro activation of purified CD4 T cells, only CCR2 was internalized, whereas expression of CXCR3 was retained in activated CD4 cells. The numbers of circulating CD4+,CXCR3+ and CD4+,CCR2+ T cells are selectively decreased during SLE flares. A decrease in the number of circulating CD4+ T cells expressing CCR2 and/or CXCR3 could serve as a biomarker of the SLE flare.

Apoptosis and expression of soluble Fas mRNA in systemic lupus erythematosus.

Our objective was to measure the relative proportions of messenger RNAs coding for soluble and membrane-bound Fas in peripheral blood mononuclear cells (PBMCs) and to quantify lymphocyte apoptosis in vitro in systemic lupus erythematosus (SLE). A RT-PCR (soluble/membrane Fas mRNA) ratio was calculated in 16 SLE, 11 RA and 16 healthy subjects' PBMCs. Apoptosis was quantitated in vitro using nick-end labeling and flow-cytometry analysis immediately and after overnight T cell activation. The mean soluble/membrane Fas ratio was 1.219+/-0.424 in the SLE group, significantly higher than in healthy subjects, 0.516+/-0.180 (P = 0.0001). There was no significant difference between inactive and active SLE groups whereas Fas ratio was higher in SLE patients with nephritis (1.460+/-0.365) compared to those without nephritis (1.031+/-0.380) (P=0.039). Mean soluble/ membrane Fas ratio in RA patients (0.975 +/- 0.37) was higher than in healthy subjects (P = 0.0003) and lower than lupus nephritis patients (P=0.015). A significantly higher proportion of mononuclear cells engaged apoptosis after T cell activation in active lupus patients, compared to control subjects. In comparison with healthy subjects, Fas alternative splicing was skewed toward the soluble form of Fas in SLE and RA. Apoptosis after T cell activation in vitro was increased in active SLE patients.

B cell derived B cell stimulatory factors in patients with systemic lupus erythematosus

We investigated the relation between polyclonal B cell activation and B cell derived B cell stimulatory factors (BSF) in patients with systemic lupus erythematosus (SLE). B-BCGF activity and IL-1 activity were detected in SAC stimulated B cells supernatant. No significant difference in BCGF-like activity was observed among the active SLE group, inactive SLE group and normal subjects. BCDF-like activity, however, was higher in the active SLE group than the inactive SLE group and normal subjects. Furthermore, IL-1 activity in the active SLE group was also higher than in the other two groups. Anti-human IL-1 antibody inhibited BCGF-like and BCDF-like activity. The inhibition rate was highest in the active SLE group. These results suggested that IL-1 production by SAC stimulated B cells was much higher in active SLE than in inactive SLE, and (that) IL-1 could act as inducer of antibody production. B cell derived IL-1 may be involved in polyclonal B cell activation. As B cells of some patients in the active SLE group spontaneously produced IL-1 and B-BCGF, we speculated that B cells had already been activated in vivo.

Investigation of B cell function and B cell stimulatory factors in patients with systemic lupus erythematosus

The immunological abnormality of systemic lupus erythematosus (SLE) is characterized by polyclonal B cell activation. Whether polyclonal B cell activation is caused by B cell abnormality or effect of B cell stimulatory factors (BSF) were investigated in this study. B cell function of SLE patients and normal individuals were measured by three methods, 1) Staphylococcus aureus Cowan I (SAC) response, 2) BCGF response and 3) BCGF response after SAC stimulation. There was no significant difference in B cell function among three groups (active SLE group, inactive SLE group and normal individuals group). Supernatants of PHA stimulated T cell and SAC stimulated B cell contained BCGF and BCDF activities. T cell derived BCGF and BCDF activities in active SLE patients revealed higher than in inactive SLE patients and normal individuals. There was no significant difference among these group in B cell derived BCGF like activity, but BCDF like activity revealed higher in active SLE patients than in inactive SLE patients and normal individuals. High level of BCGF and BCDF like activities were shown also in sera of active SLE patients found than in those of other two groups. As mentioned above, these results suggested that polyclonal B cell activation in SLE patients might be chiefly caused by increased T cell derived BCGF and BCDF activities, and B cell derived BSF respond to B cell as supportive factors.