Jesús Gómez

Enrichment of CD4+ CD25high T cell population in patients with systemic lupus erythematosus treated with glucocorticoids.

Objectives: To characterise and quantify the CD4+ CD25+ T cell population in patients with systemic lupus erythematosus (SLE) and to detect the possible influence of treatments and clinical manifestations. Methods: Characterisation of CD25low and CD25high CD4+ T cells from healthy controls and from patients with SLE was carried out using flow cytometry, analysing the expression of activation and differentiation markers. The percentage of both circulating cell subsets was determined in 56 controls and 110 unselected patients with SLE. Data were related to treatment during the past 3 months and to various clinical manifestations. Results: CD4+ CD25high lymphocytes from controls expressed low levels of CD69, CD154 or CD30, but also expressed glucocorticoid-induced tumour necrosis factor receptor, high levels of intracellular cytotoxin T lymphocyte-associated antigen 4, CD45RO and diminished amounts of CD4, all of which are phenotypic characteristics of natural regulatory T cells. CD4+ CD25low cells, on the other hand, expressed the highest levels of activation markers, indicating that they represent recently activated effector cells. Similarly, analysis of cells from patients with SLE showed the same two phenotypically distinguishable CD4+ CD25low and CD4+ CD25high populations, although both expressed slightly increased levels of activation markers. Quantitative analysis showed a considerably raised percentage of CD25low and, especially, CD25high cells in patients with SLE compared with controls. This increment was unrelated to clinical manifestations, but correlated with glucocorticoid treatment. Patients treated with glucocorticoids presented raised levels of CD25high cells, whereas untreated patients and those with anti-malarial or immunosuppressive drugs had levels similar to those in controls. Conclusions: The percentage of CD4+ CD25high cells was not altered in non-steroid-treated patients, whereas glucocorticoid treatment increased their frequency in patients with SLE.

Systemic lupus erythematosus in Asturias, Spain: clinical and serologic features.

Abstract: Asturias is an autonomous region in the north of Spain with historical and anthropologic peculiarities. In the current report, we examine the main clinical and immunologic features of 363 patients with systemic lupus erythematosus (SLE), virtually the entire population of SLE patients in Asturias. We constructed a database with the clinical and immunologic features of all patients fulfilling the American College of Rheumatology criteria, based on the review of hospital records corresponding to blood samples received for antinuclear antibodies testing since 1992. Arthritis was the most frequently observed main clinical feature and neuropathy was the rarest. Male patients had a disease more frequently characterized by serositis (p < 0.05) and neurologic disorder (p < 0.01) than females, while children presented malar rash (p < 0.05), fever (p < 0.05), and kidney involvement (p < 0.01) more often than adults. Late-onset patients were characterized by lower frequencies of malar rash (p < 0.01), neurologic disorder (p < 0.05), alopecia (p < 0.01), and lymphadenopathy (p < 0.05) than young adults. Numerous direct and inverse significant associations were found among clinical and immunologic features. The most relevant significant associations were neurologic disorder with lupus anticoagulant (p < 0.01); kidney involvement with serositis (p < 0.01) and DNA antibodies (p < 0.05); and thrombosis with DNA antibodies (p < 0.05), cardiolipin antibodies (p < 0.01), and lupus anticoagulant (p < 0.01). A low mortality was found in our series, although kidney involvement (p < 0.05) and cardiolipin antibodies (p < 0.05) are factors associated with poor survival. Abbreviations: ACR = American College of Rheumatology, CI = confidence intervals, dsDNA = double-stranded DNA, OR = odds ratio, RNP = ribonucleoprotein, SLE = systemic lupus erythematosus, SM = Smith, SSA = Sjögren syndrome A, SSB = Sjögren syndrome B.

Cytokine polymorphisms influence treatment outcomes in SLE patients treated with antimalarial drugs

Antimalarial agents have been widely used as disease-modifying antirheumatic drugs in the treatment of systemic lupus erythematosus (SLE) and other rheumatological diseases, although their mechanism of action has not yet been fully defined. It is known, however, that effective response to treatment is variable among patients. Thus, the identification of genetic predictors of treatment response would provide valuable information for therapeutic intervention. The aim of the present study was to analyze the effect of antimalarial treatment on tumor necrosis factor (TNF)α serum levels and evaluate the possible influence of TNFα and IL-10 functional genetic polymorphisms on the response to antimalarial drugs. To this end, TNFα serum levels were quantified in 171 SLE patients and 215 healthy controls by ELISA techniques and polymorphisms at positions -1,082 and -308 of the IL-10 and TNFα gene promoterswere determined by PCR amplification followed by hybridization with fluorescent-labeled allele-specific probes in 192 SLE patients and 343 matched controls. Data were related to clinical features and treatment at the time of sampling and during the course of the disease. Results showed a significantly higher amount of serum TNFα in the entire SLE population compared with controls. However, TNFα serum levels correlated negatively with the use of antimalarial treatment during at least three months before sampling. Patients under single or combined treatment with these drugs had TNFα serum levels similar to healthy controls, whereas untreated patients and those under corticosteroid or immunosuppressive therapies had increased amounts of this cytokine. This suggests, however, that antimalarial-mediated inhibition of TNFα was only significant in patients who were genetically high TNFα or low IL-10 producers. In addition, evaluation of SLE patients administered antimalarial drugs for three or more years who did not require any other specific SLE treatment indicates that patients with the combined genotype low IL-10/high TNFα are the best responders to antimalarial therapy, developing mild disease with a good course under this treatment. In conclusion, we proposed that an antimalarial-mediated downregulation of TNFα levels in SLE patients is influenced by polymorphisms at IL-10 and TNFα promoters. Our results may thus find important clinical application through the identification of patients who are the most likely to benefit from antimalarial therapy.

Glucocorticoids enhance Th17/Th1 imbalance and signal transducer and activator of transcription 3 expression in systemic lupus erythematosus patients

OBJECTIVE To investigate the relative amounts of Th17 and Th1 cells present in SLE patients and the possible effects of treatments or disease features on these populations. METHODS Peripheral blood mononuclear cells were collected from 75 SLE patients and 19 healthy controls and the proportion of Th17 and Th1 populations were assessed by flow cytometry measuring the amount of IL-17 and IFN-γ-producing cells. Gene expression of signal transducers and activators of transcription 3 (STAT3), STAT4, IL-6R and IL-12R were determined in 30 patients and 8 healthy individuals by real-time RT-PCR. Data were related to clinical and immunological parameters and to the treatment followed during the past 3 months. RESULTS Th17 cells and the Th17/Th1 ratio were significantly increased in SLE patients treated with glucocorticoids compared with healthy individuals, untreated patients or those under other treatments. No association was detected with clinical parameters, but patients with anti-ENA antibodies also displayed increased Th17 responses. Disease activity (SLEDAI) is associated with the Th17/Th1 index only in glucocorticoid-treated patients. In line with these results, gene expression of STAT3 and IL-6R was up-regulated in patients taking these drugs. Accordingly, we found a positive correlation between the Th17/Th1 ratio and STAT3 levels. CONCLUSIONS The present work provides the first evidence that aberrant Th17/Th1 balance in SLE is linked to the use of glucocorticoids and suggests that the up-regulatory effect of these drugs on the Th17 population could be associated with their ability to increase STAT3 and IL-6R expression. Additionally, anti-ENA positivity could represent a potential biomarker for Th17 bias.

Clinical associations of anti-SSA/Ro60 and anti-Ro52/TRIM21 antibodies: Diagnostic utility of their separate detection

Clinical associations of anti-SSA/Ro60 and anti-Ro52/TRIM21 antibodies are not yet fully established. In order to analyse the diagnostic utility of their separate detection, we retrospectively revised the clinical data of 200 anti-SSA/Ro60 and/or anti-Ro52/TRIM21 positive patients identified by line immunoassay during ANA routine detection. Anti-SSA/Ro60 positive patients showed a significantly higher prevalence of autoimmune diseases (AIDs) independently on the presence of anti-Ro52/TRIM21 (OR 3.13, 95% CI 1.10–8.88, p = 0.032). Anti-SSA/Ro60 was independently associated with systemic lupus erythematosus (SLE) when comparing with Sjögrens syndrome (SS) and other systemic AIDs (OR 3.46, 95% CI 1.08–11.06, p = 0.036). The more frequent specificity found in cutaneous lupus erythematosus (CLE) was also anti-SSA/Ro60. In contrast, detection of isolated anti-Ro52/TRIM21 was characteristic of SS (7/35, 20.0%), diffuse cutaneous systemic sclerosis (dcSSc) (3/4, 75.0%), primary biliary cirrhosis (PBC) (4/5, 80.0%) and, specially, of polymyositis/dermatomyositis (PM/DM) (6/6, 100%). In fact, anti-Ro52/TRIM21 was the only antibody detected in 4 out of the 6 PM/DM patients. Malignancies mainly account for the observed high prevalence of mono-specific anti-Ro52/TRIM21 in patients with non-AIDs (10/15, 62.5%). In conclusion, this retrospective study supports the routine distinction of anti-SSA/Ro60 and anti-Ro52/TRIM21 due to their different clinical associations.

Diagnostic value of anti-deamidated gliadin peptide IgG antibodies for celiac disease in children and IgA-deficient patients.

Objectives: The aim of the study was to analyze the diagnostic performance of anti-deamidated gliadin peptide (dGp) immunoglobulin (Ig) G and IgA regarding the age at celiac disease (CD) diagnosis and the anti-dGp IgG usefulness for diagnosing CD IgA-deficient patients. Methods: Anti-dGp IgG and IgA and anti-native gliadin (nGlia) IgA were determined by enzyme fluoroimmunoassay in 100 newly diagnosed anti-tissue transglutaminase (tTG) IgA-positive pediatric and adult patients with CD and in 100 age-matched patients with other digestive pathologies. Anti-dGp IgG was evaluated in 6 CD IgA-deficient patients. Results: When analyzing all of the patients, the anti-dGp IgG assay showed higher diagnostic accuracy (area under receiver operating characteristic curve), specificity, and positive predictive value than anti-dGp IgA and anti-nGlia IgA. All of the diagnostic parameters corresponding to anti-dGp IgG reached the same values as anti-tTG IgA in children 7 years or younger. In older patients, both anti-dGp isotypes showed an inverse behavior, IgG having a higher specificity and positive predictive value but a lower sensitivity and negative predictive value than IgA. Anti-dGp levels were associated with the severity of intestinal lesions, and an inverse association was found regarding age at diagnosis. Both anti-dGp IgG and IgA were found to be positive in the 9 patients with minimal intestinal changes included in the study. All of the patients with CD with IgA deficiency were positive for anti-dGp IgG. Conclusions: The diagnostic performance of anti-dGp depends on the antibody isotype and on the age at CD diagnosis, anti-dGp IgG being a serological marker at least as useful as anti-tTG IgA for detecting CD in children ages 7 years or younger. Our data also indicate that anti-dGp IgG can improve the diagnosis of IgA-deficient patients.

Behavioural and population responses to changing availability of Artemia prey by moulting black-necked grebes, Podiceps nigricollis

We examined how availability of brine shrimps, Artemia parthenogenetica, influenced temporal aspects of foraging behaviour and population dynamics of moulting black-necked grebes, Podiceps nigricollis, from late August to early December in four salt ponds in the Odiel marshes, southern Spain, in 2008 and 2009. The moulting grebe population was higher in 2009, coinciding with an increase in shrimp biomass, with a peak of 2,500 birds in October. Grebes increased their time spent foraging as the season progressed, coinciding with decreases in shrimp biomass and water temperature. Foraging activity was lower in 2009, when shrimp biomass was greater. Diving was the most frequent feeding method, especially as the season progressed. Brine shrimps at the bottom of the water column were larger than those near the surface. Differences between years in grebe body mass suggest that changing shrimp availability and water temperature had an influence on body condition. The grebe population consumed an estimated 0.2–2.0% of the standing crop of Artemia per day, with this fraction increasing as the season progresses, thus contributing to the decline in the Artemia population. Our results suggest that moulting grebes are probably only able to adjust foraging effort within certain limits, especially at the end of moulting period when thermal stress is greatest and food supply is lowest. They may leave the study area when they can no longer meet their energy requirements.

Common and Specific Associations of Anti-SSA/Ro60 and Anti-Ro52/TRIM21 Antibodies in Systemic Lupus Erythematosus

Little information exists about the association of anti-SSA/Ro60 and anti-Ro52/TRIM21 with systemic lupus erytematosus (SLE) features. In this work, we analysed the associations of both anti-Ro reactivities with clinical and immunological manifestations in 141 SLE patients. Photosensitivity and xerophtalmia/xerostomia were found to be positively associated with both anti-SSA/Ro60 (P = 0.024 and P = 0.019, resp.) and anti-Ro52/TRIM21 (P = 0.026 and P = 0.022, resp.). In contrast, a negative association was detected regarding anti-phospholipid antibodies, anti-SSA/Ro60 having a stronger effect (P = 0.014) than anti-Ro52/TRIM21. Anti-SSA/Ro60 showed a specific positive association with hypocomplementemia (P = 0.041), mainly with low C4 levels (P = 0.008), whereas anti-Ro52/TRIM21 was found to be positively associated with Raynauds phenomenon (P = 0.026) and cytopenia (P = 0.048) and negatively associated with anti-dsDNA (P = 0.013). Lymphocytes are involved in the relationship between anti-Ro52/TRIM21 and cytopenia since positive patients showed lower cell levels than negative patients (P = 0.036). In conclusion, anti-SSA/Ro60 and anti-Ro52/TRIM21 showed both common and specific associations in SLE. These data thus increase evidence of the different associations of the two anti-Ro specificities even in a particular disease.

Relationship between FOXP3 positive populations and cytokine production in systemic lupus erythematosus

In this work we studied CD4+FOXP3+ populations in systemic lupus erythematosus (SLE) and the relationship with Th cytokine production. We found an increment in CD25-FOXP3+ population in SLE associated with CD4+ downregulation and disease progression. CD25low cells were also upregulated and showed increased percentages of FOXP3+ and CD127-/low cells, supporting the activated status of SLE lymphocytes. Despite the normal levels of CD25highFOXP3+ cells, the negative correlations observed in controls with the frequency of IFNγ, TNFα and IL-10 secreting cells were disrupted in patients, supporting a defective Treg function. Also, CD25high cells showed an altered balance in the production of these cytokines. In addition, CD25highFOXP3+ cells correlated directly with IL-17A and IL-8 but not with TGFβ in SLE. The increased proportion of IL-17+ cells among the CD25high subset and the positive correlation between IL-17 levels and Treg cells suggest a trans-differentiation of Treg into Th17 cells in SLE.

Antibodies to mitotic spindle apparatus: clinical significance of NuMA and HsEg5 autoantibodies.

IntroductionThe clinical associations of NuMA and HsEg5 antibodies, the main anti-mitotic spindle apparatus autoantibodies, remain unclear due to their extremely low prevalence.Patients and MethodsWe have analysed the clinical data of 40 anti-NuMA- (0.87‰) and 7 anti-HsEg5- (0.15‰) positive patients detected during routine immunofluorescence examination of 45,804 sera. NuMA reactivity was further confirmed by immunoblotting.ResultsAntibodies to HsEg5 did not associate with any specific pathology. NuMA positivity associated with a diagnosis of connective tissue disease (CTD) in 18 patients (45%), primary Sjögren or sicca syndrome and undifferentiated connective tissue disease being the most represented. Seven patients (17.5%) were diagnosed with different organ-specific autoimmune diseases, whereas in the other 15 patients (37.5%), no autoimmune pathology could be documented.ConclusionsTherefore, although both anti-mitotic spindle apparatus antibodies are not associated to a defined autoimmune pathology, the presence of NuMA antibodies, mainly at high titers, may be an indication for a more extensive screening of CTD.