Adriana Lucía Vanegas

Asociación entre títulos de anticuerpos anticardiolipinas y eventos trombóticos

Resumen Introduccion la trombosis es la manifestacion principal del sindrome antifosfolipido (SAF); los marcadores serologicos de esta entidad son los anticuerpos anticardiolipinas (aCL), la anti-s2 glicoproteina 1 y el anticoagulante lupico. Aun se discute si los titulos de aCL o la presencia de un “segundo hit” son factores de riesgo para trombosis. Objetivo evaluar la asociacion entre fenomenos tromboticos vasculares con la presencia y los titulos de aCL; ademas del papel de otros factores protromboticos. Material y metodo estudio descriptivo transversal. Se revisaron historias clinicas de pacientes con sospecha clinica de SAF y con al menos una medicion de titulos de aCL, se evaluo la presencia o no de eventos tromboticos y de comorbilidades (segundo hit). Resultados historias clinicas de 49 pacientes, 33 con un total de 36 eventos tromboticos de los cuales 23 ocurrieron en lechos venosos y 13 en lechos arteriales. Aunque la mayoria de los pacientes con titulos de aCL > 20 GLP o MLP se encontraban en el grupo de trombosis, no se encontro asociacion significativa entre la presencia de trombosis y los titulos de aCL; como tampoco entre trombosis y la existencia de otras comorbilidades. Conclusiones los hallazgos encontrados permiten sugerir la mayor frecuencia de eventos tromboticos en pacientes con titulos de aCL

Potential Involvement of Platelet-Derived Microparticles and Microparticles Forming Immune Complexes during Monocyte Activation in Patients with Systemic Lupus Erythematosus

Microparticles (MPs) are vesicles derived from the plasma membrane of different cells, are considered a source of circulating autoantigens, and can form immune complexes (MPs-ICs). The number of MPs and MPs-ICs increases in patients with systemic lupus erythematosus (SLE). MPs activate myeloid cells by inducing IL-6 and TNF-α in both SLE and other diseases. Therefore, we propose that the recognition of MPs-ICs by monocytes rather that MPs may define their phenotype and contribute to the inflammatory process in patients with SLE. Thus, the aims of this study were to evaluate the association among circulating MPs-ICs from different cell sources, alterations observed in monocyte subsets, and disease activity in patients with SLE and to establish whether monocytes bind and respond to MPs-ICs in vitro. Circulating MPs and monocyte subsets were characterized in 60 patients with SLE and 60 healthy controls (HCs) using multiparametric flow cytometry. Patients had higher MP counts and frequencies of MPs-CD41a + (platelet-derived) compared with HCs, regardless of disease activity. MPs from patients with SLE were C1q + and formed ICs with IgM and IgG. MPs-IgG + were positively correlated with active SLE (aSLE), whereas MPs-IgM + were negatively correlated. Most of the circulating total ICs-IgG + were located on MPs. The proportion and number of non-classical monocytes were significantly decreased in patients with SLE compared with HCs and in patients with aSLE compared with patients with the inactive disease. Non-classical monocytes obtained from patients with SLE exhibited increased levels of CD64 associated with MPs-IgG +, MPs-C1q +, total circulating ICs-IgG +, and disease activity. The direct effects of MPs and MPs-IgG + on monocytes were evaluated in cell culture. Monocytes from both HCs and patients bound to and internalized MPs and MPs-IgG + independent of CD64. These vesicles derived from platelets (PMPs), mainly PMPs-IgG +, activated monocytes in vitro and increased the expression of CD69, CD64, and pro-inflammatory cytokines such as IL-1β, TNF-α, and IFN-α. Therefore, MPs are one of the most representative sources of the total amount of circulating ICs-IgG + in patients with SLE. MPs-IgG + are associated with SLE activity, and PMPs-IgG + stimulate monocytes, changing their phenotype and promoting pro-inflammatory responses related to disease activity.

Infiltrating CD16+ Are Associated with a Reduction in Peripheral CD14+CD16++ Monocytes and Severe Forms of Lupus Nephritis

Our aim was to characterize glomerular monocytes (Mo) infiltration and to correlate them with peripheral circulating Mo subsets and severity of lupus nephritis (LN). Methods. We evaluated 48 LN biopsy samples from a referral hospital. Recognition of Mo cells was done using microscopic view and immunohistochemistry stain with CD14 and CD16. Based on the number of cells, we classified LN samples as low degree of diffuse infiltration (<5 cells) and high degree of diffuse infiltration (≥5 cells). Immunophenotyping of peripheral Mo subsets was done using flow cytometry. Results. Mean age was 34.0 ± 11.7 years and the mean SLEDAI was 17.5 ± 6.9. The most common SLE manifestations were proteinuria (91%) and hypocomplementemia (75%). Severe LN was found in 70% of patients (Class III, 27%; Class IV, 43%). Severe LN patients and patients with higher grade of CD16+ infiltration had lower levels of nonclassical (CD14+CD16++) Mo in peripheral blood. Conclusions. Our results might suggest that those patients with more severe forms of LN had a higher grade of CD14+CD16+ infiltration and lower peripheral levels of nonclassical (CD14+CD16++) Mo and might reflect a recruitment process in renal tissues. However, given the small sample, our results must be interpreted carefully.

Tuberculous sacroiliitis in a patient with systemic lupus erythematosus: a case report and literature review:

Systemic lupus erythematosus (SLE) patients are at higher risk of developing opportunistic infections such as tuberculosis (TB), especially extrapulmonary forms like osteoarticular TB, compared to the general population. However, tuberculous sacroiliitis has been scarcely reported in these patients. We present a 34-year-old woman with SLE who developed articular tuberculosis simultaneously affecting the right sacroiliac joint and the left knee. The patient was successfully treated with antituberculosis therapy for nine months. In this case, in addition to the immunological abnormalities of lupus, the long-term glucocorticoid therapy at high dosages was the main risk factor for the development of osteoarticular tuberculosis.

273 Urinary tweak levels as biomarker of lupus nephritis in colombian sle patients

Background and aims TNF-like WEAK inducer of apoptosis (TWEAK), a TNF ligand superfamily is mainly produced by monocytes/macrophages, and is widely expressed at the RNA level in tissues including kidneys. The usefulness of urinary TWEAK (uTWEAK) to identify renal involvement in Mestizo and African-Latin American (ALA) SLE patients has not been examined yet. Methods Patients meeting the revised ACR criteria for SLE were recruited from 2 different centres at Medellín and Baranquilla, Colombia. uTWEAK were measured using an ELISA kit (R and D system, USA) Results 158 SLE patients were recruited (89% female) with median age of 32.8±12.1 years and median disease duration of 7.27±6.6 years. Mestizo (77%) and ALA (20%) were majority. 64% of patients had lupus nephritis (LN). 50 out of 71 biopsy proven LN had proliferative forms. Mean SLEDAI score was 8.5±8.7. LN patients (2803±6086 vs 672±1042, p=0.013) (Fig 1A) and ALA patients (3995±9656 vs 1618±2653, p=0.002) had significant higher levels of uTWEAK. uTWEAK levels were higher in patients with active LN and in Class V LN (Fig. 1B). uTWEAK levels were significantly correlated with 24 hours proteinuria, SLEDAI (Fig. 1C) and serum anti-C1q titers. An ROC curve constructed showed a good level of sensitivity and specificity (Fig. 1D) Abstract 273 Figure 1 Conclusions In our cohort of Colombian SLE patients, uTWEAK levels were 4 and 2 times higher in LN patients and ALA respectively. uTWEAK were significantly higher in active LN and were correlated with disease activity, proteinuria and anti-C1q antibodies.

274 Prevalence of anti-dfs70 antibodies in a colombian cohort: a case-control study

Background and aims Anti-dense fine speckled 70 (anti-DFS70) antibodies were initially identified as an ANA IIF pattern from patients with interstitial cystitis; however, some recent studies showed that anti-DFS70 antibodies are common among ANA positive individuals with no evidence of systemic autoimmune disease (SAD)(Mahler M. 2012). Information of anti-DFS70 in Latin-American countries is very limited. We determined the prevalence of Anti-DFS70 antibodies in a Colombian cohort. Methods We evaluated individuals≥18 years old, including 100 SLE patients, 102 SADs, 200 healthy controls, and 56 subjects suspected of having autoimmune disease with ANA positive and negative anti ds-DNA antibodies. The presence of anti-DFS70 antibodies was determined by QUANTA Flash by chemiluminescent techniques (Inova/Werfen, San Diego) Results Our final cohort included 458 samples. The mean age of SLE patients was 33±12 years, for SADs was 41±19 and for healthy controls was 36±10 years. The main diagnoses of SAD were: Vasculitis (n=28), RA (n=21), Systemic sclerosis (n=12), primary antiphospholipid syndrome (n=11), dermatomyositis (n=10) among others. Racial/ethnic breakdown was: 76% Mestizo and 20% Afro-latin Americans. Anti-DFS70 antibodies were positive in 1.8% of subjects with ANAs positive/anti DNA negative, in 1% of SLE patients, 0.9% of patients with other SADs and in 0.5% of healthy controls. Given the low prevalence of anti-DFS70 antibodies, no clinical correlations were possible. Conclusions Despite anti-DFS70 antibodies are a good diagnostic tool for discrimination among healthy individuals and SADs (including SLE), we found a very low prevalence of anti-DFS70 antibodies in our Colombian cohort.

Síndrome de seudotumor cerebral en una mujer embarazada y con lupus eritematoso sistémico

Systemic lupus erythematous is a chronic multi-systemic autoimmune disease that affects multiple organ systems, including the central nervous system. Pseudotumor cerebri is a disorder associated with increased intracranial pressure in the absence of a space-occupying lesion or other identifiable cause that affects young and obese women.We present the case of a pregnant woman with both pseudotumor cerebri and a new diagnosis of active systemic lupus erythematous.

CE-06 Urinary neutrophil gelatinase – associated lipocalin and monocyte chemoattractant protein-1 as biomarkers for lupus nephritis in colombian SLE patients

Background Some previous studies in Caucasian, Asian, and African-American patients have shown that urine levels of Neutrophil Gelatinase–Associated Lipocalin (uNGAL) and Monocyte Chemoattractant Protein-1 (uMCP-1) were significantly greater in patients with LN. However, information in Mestizo and Afro-Latin American patients is very limited. Our aim was to evaluate diagnostic value of uNGAL and uMCP-1 as potential markers for the diagnosis of LN in Colombian SLE patients Materials and methods We examined urinary levels of NGAL and MCP-1 in 93 consecutive SLE patients (ACR criteria 1982) from Hospital San Vicente Fundación, at Medellín, Colombia. uNGAL and uMCP-1 were measured by ELISA techniques (R&D system, Minneapolis, USA). In addition, serum Anti-C1q antibodies were measured (Inova, San Diego, USA). Several clinical and serological features were analysed as well as disease activity (SLEDAI). Mann-Whitney tests were used to compare data and Spearman’s rho for correlations. Additionally, ROC curves relating the specificity and sensitivity profiles of the 2 biomarkers were done. Results Ninety-three SLE patients were recruited (88% female) with median age of 33.6 ± 12.4 years and median disease duration of 11.5 ± 14.8 years. Mestizo (75%) and Afro-Latin American (22%) were majority. One quarter of patients had an early SLE (< 2 years of duration) and 64 were admitted at the time of urine collection. Hematologic disease (89%), arthritis (83%), cutaneous involvement (82%), and renal disease (66%) were among most common manifestations. 63% of patients were positive for anti-C1q. We found significant positive correlation between uNGAL levels and SLEDAI (r = 0.331, p = 0.02) and between uMCP1 with SLEDAI (r = 0.428, p < 0.02) and with uNGAL (r = 0.467, p < 0.0001). uNGAL and uMCP-1 were significantly higher in patients with LN than in patients without LN (53.0 ± 56.3 vs 16.0 ± 16.6 pg/ml, p = 0.001 and 2340.4 ± 4521.4 vs 472.4 ± 596.5, p = 0.015, respectively). uNGAL levels were also significantly higher in patients with active LN (>500 mg proteinuria/24 hrs) than in inactive LN (66.1 ± 61.9 vs 9.0 ± 8.6, p < 0.001). A ROC curve constructed for uNGAL, uMCP-1, and anti-C1q for LN in all SLE patients showed a good level of sensitivity and specificity (Figure 1). Conclusions Colombian LN patients had 4 times and 5 times higher levels of uNGAL and uMPC-1, respectively than patients without LN. Additionally, uNGAL was significantly higher in patients with active LN. Both markers were correlated with disease activity. A multinational prospective study is ongoing under GLADEL cohort, in order to evaluate those biomarkers in 14 Latin American countries. Abstract CE-06 Figure 1 Receiver operating characteristic (ROC) curve of urinary NGAL, MCP-1 and anti C1q for the identification of LN (dotted line for uNGAL, solid green line for uMCP-1 and solid red line for anti C1q) Acknowledgements JA Gómez-Puerta was supported by Colciencias (conv. 656 de 2014). Anti-C1q antibodies were provided by Inova, Werfen, Colombia

THU0335 Anti C1-Q Antibodies and Urinary Monocyte Chemoattractant Protein-1 Levels as Biomarkers for Renal Involvement in Patients with Systemic Lupus Erythematosus

Background Some previous studies in Caucasian, Asian, and African-american patients have shown that the anti-C1q antibodies and urine levels of MCP-1 (uMCP-1) were significantly greater in patients with LN. However, information in Mestizo and Afro-Latin American patients is very limited. Objectives Our aim was to evaluate diagnostic value of Anti-C1q and uMCP-1 as a potential markers for the diagnosis of LN in Colombian SLE patients Methods We examined serum levels of anti-C1q and uMCP-1 in 67 consecutive SLE patients (ACR criteria 1997) from Hospital San Vicente Fundaciόn, at Medellín, Colombia. A fresh urine sample from each patient was collected in a sterile container. Serum Anti-C1q antibodies and uMCP-1 were measured by ELISA techniques (Inova, San Diego, and R&D system, Minneapolis, EEUU, respectively). Several clinical and serological features were analyzed as well as disease activity (SLEDAI). Descriptive statistics were used to describe data. Mann-Whitney tests were used to compare data and Spearmansrho for correlations. Additionally, ROC curves relating the specificity and sensitivity profiles of the 2 biomarkers were done Results 67 SLE patients were recruited (89% female) with median age of 33.5± 12.0 yrs and median disease duration of 6.76 ± 6.80 years. Mestizo (71%) and Afro-latin American (26%) were majority. Hematologic disease (89%), arthritis (86%), cutaneous involvement (80%), and renal disease (66%) were among most common manifestations. Sixty-three percent of patients were positive for anti-C1q and 56% had elevated levels of uMCP-1 (mean levels 1678 ±3722 pg/ml). Mean SLEDAI score was 10.0 ± 9.0. We found significant association between anti C1q antibodies and several clinical features including serositis, proteinuria, renal and hematological involvement and with several serological markers including anti-dsDNA, low complement and higher SLEDAI score. Patients with LN had higher levels of anti-C1q antibodies than patients without LN (mean levels of 91.3 ± 85.4 vs 31.7 ± 38.7, p value=0.002). uMCP-1 levels were significantly higher in patients with LN than in patients without LN (2399 ± 4566 vs 472 ± 596 pg/ml, p value=0.017) and in patients with proteinuria (2537 ± 4731 vs 553 ± 688, p value=0.025). There was no correlation between anti-C1q levels and uMCP-1. An ROC curve constructed for anti-C1q and uMCP-1 and renal in all SLE patients showed that uNGAL had a good level of sensitivity and specificity with an AUC of 0.74 for anti-C1q and 0.82for uMCP1 (see figure). ROC curve on Anti-C1q and uMCP-1. Solid line for Anti-C1q, dotted line for MCP-1. Conclusions Although further confirmatory testing is underway, LN patients had almost 3 times and 5 times higher levels of Anti-C1q and uMCP-1, respectively than patients without LN. Anti-C1q antibodies and uMCP-1 may be a good biomarkers for LN in Mestizo and Afro-Latin american SLE patients, and can be a useful marker to identify patients with higher risk of renal involvement Acknowledgement JA Gόmez-Puerta was supported by Colciencias (conv. 656 de 2014). Anti-C1q antibodies were provided by Inova, Werfen, Colombia Disclosure of Interest None declared

Smad y otros blancos terapéuticos en esclerodermia

Resumen La esclerodermia es una enfermedad caracterizada por la acumulacion excesiva de tejido fibroso que lleva a alteracion en la estructura y funcion de la piel y de organos internos. La principal citoquina involucrada en este proceso es el factor transformante de crecimiento beta y sus funciones se realizan principalmente a traves de la senalizacion intracelular mediada por las proteinas Smad. Se han desarrollado estrategias para bloquear los efectos del factor transformante de crecimiento beta y la identificacion de la via de transmision de senales proporciona nuevas herramientas para la investigacion de futuras terapias, pero son necesarios mas estudios en modelos animales y en humanos que logren reproducir en forma satisfactoria y segura los resultados. El proposito de este articulo es analizar la funcion del factor transformante de crecimiento beta en la fisiopatologia de la esclerodermia profundizando en la via de senalizacion mediada por Smad; ademas, revisar los estudios que involucran estas proteinas como blanco terapeutico de moleculas y medicamentos como posibles tratamientos para la esclerodermia.