Comparative study of the glistening between four intraocular lens models assessed by OCT and deep learning.

PURPOSE: To evaluate the glistening in 4 different models of intraocular lenses (IOLs) using optical coherence tomography (OCT) and deep learning (DL). SETTING: Centro Internacional de Oftalmología Avanzada (Madrid, Spain). DESIGN: Cross-sectional study. METHODS: 325 eyes were assessed for the presence and severity of glistening in 4 IOL models: ReSTOR+3 SN6AD1 (n = 41), SN60WF (n = 110), PanOptix TFNT (n = 128) and Vivity DFT015 (n = 46). The presence of glistening was analyzed using OCT, identifying the presence of hyperreflective foci (HRF) in the central area of the IOL. A manual and an original DL-based quantification algorithm designed for this purpose was applied. RESULTS: Glistening was detected in 22 (53.7%) ReSTOR SN6AD1, 44 (40%) SN60WF, 49 (38.3%) PanOptix TFNT, and 4 (8.7%) Vivity DFT015 IOLs, when any grade was considered. In the comparison of the different types of IOLs, global glistening measured as total HRF was 17.3 ± 25.9 for the ReSTOR+3; 9.3 ± 15.7 for the SN60WF; 6.9 ± 10.5 for the PanOptix; and 1.2 ± 2.6 for the Vivity ( P < .05). There was excellent agreement between manual and DL-based quantification (≥0.829). CONCLUSIONS: It is possible to quantify, classify and compare the glistening severity in different IOL models using OCT images in a simple and objective manner with a DL algorithm. In the comparative study, the Vivity presented the lowest severity of glistening.

Does imbalance in chest X-ray datasets produce biased deep learning approaches for COVID-19 screening?

BACKGROUND: The health crisis resulting from the global COVID-19 pandemic highlighted more than ever the need for rapid, reliable and safe methods of diagnosis and monitoring of respiratory diseases. To study pulmonary involvement in detail, one of the most common resources is the use of different lung imaging modalities (like chest radiography) to explore the possible affected areas. METHODS: The study of patient characteristics like sex and age in pathologies of this type is crucial for gaining knowledge of the disease and for avoiding biases due to the clear scarcity of data when developing representative systems. In this work, we performed an analysis of these factors in chest X-ray images to identify biases. Specifically, 11 imbalance scenarios were defined with female and male COVID-19 patients present in different proportions for the sex analysis, and 6 scenarios where only one specific age range was used for training for the age factor. In each study, 3 different approaches for automatic COVID-19 screening were used: Normal vs COVID-19, Pneumonia vs COVID-19 and Non-COVID-19 vs COVID-19. The study was validated using two public chest X-ray datasets, allowing a reliable analysis to support the clinical decision-making process. RESULTS: The results for the sex-related analysis indicate this factor slightly affects the system in the Normal VS COVID-19 and Pneumonia VS COVID-19 approaches, although the identified differences are not relevant enough to worsen considerably the system. Regarding the age-related analysis, this factor was observed to be influencing the system in a more consistent way than the sex factor, as it was present in all considered scenarios. However, this worsening does not represent a major factor, as it is not of great magnitude. CONCLUSIONS: Multiple studies have been conducted in other fields in order to determine if certain patient characteristics such as sex or age influenced these deep learning systems. However, to the best of our knowledge, this study has not been done for COVID-19 despite the urgency and lack of COVID-19 chest x-ray images. The presented results evidenced that the proposed methodology and tested approaches allow a robust and reliable analysis to support the clinical decision-making process in this pandemic scenario.

Pregnancy and systemic lupus erythematosus in Spain: Has anything changed in the 21st century?

OBJECTIVE: To analyse a cohort of pregnant patients with systemic lupus erythematosus and compare the outcomes of both the disease and pregnancy with the results of previous studies conducted in the same geographical area. PATIENTS AND METHODS: Retrospective cohort study of 37 women with systemic lupus erythematosus (64 pregnancies) followed in a multidisciplinary unit. Comparative study with similar Spanish studies identified after literature search. RESULTS: Our cohort was characterized by an older age and by the presence of non-Caucasian patients. Although we found no clinical differences, from the serological point of view our cohort presented a higher frequency of antiphospholipid antibodies. Patients included in this study were treated more frequently with antimalarials and low-dose aspirin. Systemic lupus erythematosus flare frequency was very similar between the different studies, and we did not identify clear predictors for them. Although the rate of live births was similar among studies, the obstetric outcome of our series was better with a very low rate of preeclampsia, preterm birth and low birth weight newborn. The only predictor of adverse obstetric event was age. CONCLUSIONS: Although changes in the therapeutic attitude and planning of pregnancy in recent years have not had a direct impact on the rate of systemic lupus erythematosus flares during pregnancy, they have meant an improvement in the obstetric results. The introduction of new variables independent of the disease such as age at conception, socio-cultural origin, or the availability of multidisciplinary units should be considered in the results of future studies.

Embarazo y lupus eritematoso sistémico en España: ¿ha cambiado algo en el siglo xxi? / Pregnancy and Systemic Lupus Erythematosus in Spain: Has Anything Changed in the 21st Century?

Objetivo: Analizar una cohorte de pacientes embarazadas con lupus eritematoso sistémico y comparar los desenlaces tanto de la enfermedad como del embarazo con los resultados de estudios previos realizados en la misma área geográfica. Pacientes y métodos: Estudio de cohortes retrospectivo de 37 mujeres con lupus eritematoso sistémico (64 embarazos) seguidas en una consulta multidisciplinar. Estudio comparativo con los estudios españoles similares identificados tras revisión bibliográfica. Resultados: Nuestra cohorte se caracterizó por una edad más elevada y por la presencia de pacientes de origen no caucásico. Aunque no encontramos diferencias clínicas relevantes, serológicamente nuestra cohorte presentó una mayor frecuencia de anticuerpos antifosfolípido. Las pacientes incluidas en este estudio fueron tratadas más frecuentemente con antipalúdicos y aspirina. La frecuencia de brotes fue muy similar entre los distintos estudios, y no identificamos predictores claros para los mismos. Aunque la tasa de nacidos vivos fue similar, el desenlace obstétrico de nuestra serie fue mejor, con una baja tasa de preeclampsia, parto pretérmino y recién nacido de bajo peso. El único predictor de acontecimiento obstétrico adverso fue la edad. Conclusiones: Si bien los cambios en la actitud terapéutica y la planificación del embarazo no han tenido un impacto directo sobre la tasa de reactivación del lupus eritematoso sistémico durante el embarazo, sí que han supuesto una mejoría en los resultados obstétricos. La introducción de nuevas variables independientes de la enfermedad como la edad en la concepción, la procedencia sociocultural, o la disponibilidad de unidades multidisciplinares deberán ser consideradas en los resultados de próximos estudios.(AU)

Objective: To analyse a cohort of pregnant patients with systemic lupus erythematosus and compare the outcomes of both the disease and pregnancy with the results of previous studies conducted in the same geographical area. Patients and methods: Retrospective cohort study of 37 women with systemic lupus erythematosus (64 pregnancies) followed in a multidisciplinary unit. Comparative study with similar Spanish studies identified after literature search. Results: Our cohort was characterized by an older age and by the presence of non-Caucasian patients. Although we found no clinical differences, from the serological point of view our cohort presented a higher frequency of antiphospholipid antibodies. Patients included in this study were treated more frequently with antimalarials and low-dose aspirin. Systemic lupus erythematosus flare frequency was very similar between the different studies, and we did not identify clear predictors for them. Although the rate of live births was similar among studies, the obstetric outcome of our series was better with a very low rate of preeclampsia, preterm birth and low birth weight newborn. The only predictor of adverse obstetric event was age. Conclusions: Although changes in the therapeutic attitude and planning of pregnancy in recent years have not had a direct impact on the rate of systemic lupus erythematosus flares during pregnancy, they have meant an improvement in the obstetric results. The introduction of new variables independent of the disease such as age at conception, socio-cultural origin, or the availability of multidisciplinary units should be considered in the results of future studies.(AU)

Optimal large-scale CD34+ enrichment from a leukapheresis collection using the clinimacs prodigy platform.

Optimization of Hematology Patient's treatment: It is possible to obtain a 100% CD34+ recovery after CD34+ selection using the CliniMACS Prodigy.

Pregnancy and Systemic Lupus Erythematosus in Spain: Has Anything Changed in the 21st Century? / Embarazo y lupus eritematoso sistémico en España: ¿ha cambiado algo en el siglo xxi?

OBJECTIVE: To analyse a cohort of pregnant patients with systemic lupus erythematosus and compare the outcomes of both the disease and pregnancy with the results of previous studies conducted in the same geographical area. PATIENTS AND METHODS: Retrospective cohort study of 37 women with systemic lupus erythematosus (64 pregnancies) followed in a multidisciplinary unit. Comparative study with similar Spanish studies identified after literature search. RESULTS: Our cohort was characterized by an older age and by the presence of non-Caucasian patients. Although we found no clinical differences, from the serological point of view our cohort presented a higher frequency of antiphospholipid antibodies. Patients included in this study were treated more frequently with antimalarials and low-dose aspirin. Systemic lupus erythematosus flare frequency was very similar between the different studies, and we did not identify clear predictors for them. Although the rate of live births was similar among studies, the obstetric outcome of our series was better with a very low rate of preeclampsia, preterm birth and low birth weight newborn. The only predictor of adverse obstetric event was age. CONCLUSIONS: Although changes in the therapeutic attitude and planning of pregnancy in recent years have not had a direct impact on the rate of systemic lupus erythematosus flares during pregnancy, they have meant an improvement in the obstetric results. The introduction of new variables independent of the disease such as age at conception, socio-cultural origin, or the availability of multidisciplinary units should be considered in the results of future studies.

Altered Th17/Treg Ratio in Peripheral Blood of Systemic Lupus Erythematosus but Not Primary Antiphospholipid Syndrome.

Introduction: The role of the immune response in the pathogenesis of antiphospholipid syndrome (APS) remains elusive. It is possible that differences in the frequencies of Th17 cells and/or defects in the immunoregulatory mechanisms are involved in the pathogenesis of APS. Our aim was to determine the peripheral blood Th cells phenotype and the circulating cytokine profile in patients with primary APS (pAPS) and compare it with systemic lupus erythemathosus (SLE) as disease control group. Methods: The frequencies of circulating regulatory T cells (Tregs) were determined in PBMCs from 36 patients with pAPS by flow cytometry. As control groups we included 21 age- and gender-matched healthy controls (HC) and 11 patients with SLE. The suppressive capacity of Tregs was evaluated in vitro by coculture assay. On the other hand, intracellular cytokine production was assessed in Th1, Th2, and Th17 cells and circulating IL-6, IL-10, and IL-35 were measured by Cytometric Bead Array and ELISA. The quantification of Th master gene expression levels was performed by real time quantitative PCR. Results: pAPS patients and SLE patients did not show differences in the percentage or number of Tregs compared to HC. The suppressive capacity of Tregs was also similar in the three study group. Instead, we found higher FoxP3·mRNA expression levels in pAPS patients and HC than SLE patients. Regarding the Th17 response, patients with pAPS and HC showed a significantly lower frequency of circulating Th17 cells than SLE. However, no differences were observed in the Th1 response between patients and controls. Thus, increased Th17/Th1 and Th17/Treg ratios were found in SLE patients but not in pAPS patients. pAPS and SLE patients had higher serum IL-6 levels than HC but there was not difference between both disease groups. Besides, a significant increase in the immunosuppressive cytokine levels was observed only in pAPS as compared to HC. Conclusions: Our data demonstrate an increased inflammatory profile of peripheral blood CD4+ T cells from SLE as compared with pAPS mostly due to an increased Th17 response. In conclusion, there seems not to be a direct pathogenic role for Th cells in pAPS but in SLE.

Peripheral B-Cell Subset Distribution in Primary Antiphospholipid Syndrome.

BACKGROUND: B-cell differentiation and B-cell tolerance checkpoints may be different in antiphospholipid syndrome (APS) from systemic lupus erythematosus (SLE) and can help to understand differences between them. Our aim was to define alterations of B-cell subsets in patients with primary APS (pAPS) and to compare them with SLE patients and healthy controls (HC). METHODS: Cross-sectional study including three study groups: 37 patients with pAPS, 11 SLE patients, and 21 age- and gender-matched HC. We determined the frequencies of different B-cell subsets in peripheral blood naïve and memory compartments. In addition, we measured serum B cell-activating factor (BAFF) levels and circulating pro-inflammatory cytokines, such as IL-6, by commercial ELISA and CBA, respectively. RESULTS: Patients with pAPS showed a lower percentage of immature and naïve B cells than patients with SLE (p = 0.013 and p = 0.010, respectively) and a higher percentage of non-switched memory B cells than patients with SLE (p = 0.001). No differences either in the percentage of switched memory cells or plasma cells were found among the different groups. Serum BAFF levels were higher in SLE patients than in healthy controls and pAPS patients (p = 0.001 and p = 0.017, respectively). A significant increase in the serum BAFF levels was also observed in pAPS patients compared to HC (p = 0.047). Circulating IL-6 levels were higher in SLE and pAPS patients than HC (p = 0.036 and p = 0.048, respectively). A positive correlation was found between serum BAFF and IL-6 levels in patients with SLE but not in pAPS (p = 0.011). CONCLUSIONS: Our characterization of peripheral blood B-cell phenotypes in pAPS demonstrates different frequencies of circulating B cells at different stages of differentiation. These differences in the naïve B-cell repertoire could explain the higher number and variety of autoantibodies in SLE patients in comparison to pAPS patients, especially in those with obstetric complications.

Evaluation of Toll-like-receptor gene family variants as prognostic biomarkers in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disease whose main feature is persistent joint inflammation. Toll-like receptors (TLRs) play critical roles in the activation of innate and adaptive immune responses, and influence the activity of NFκB, a key player in chronic inflammation. We aimed at investigating the association of TLR allelic variants with susceptibility and severity of RA through a systematic, high-throughput, analysis of TLR genes. All coding exons and flanking regions of nine members of the TLR family (TLR1-9) were analyzed in 66 patients with RA and 30 healthy controls by next generation sequencing. We focussed on three single allelic variants, N248S in TLR1, Q11L in TLR7 and M1V in TLR8 based on the allelic frequencies in both patient and control populations, the predicted impact on protein function and the novelty in RA research. Analysis of these selected variants in a larger cohort of 402 patients with RA and in 208 controls revealed no association with susceptibility. However, the M1V allele was associated with a lower need for disease-modifying antirheumatic drugs (DMARDs) (p=0.008) and biologic treatments (p=0.021). Functional studies showed that the M1V variant leads to a reduced production of inflammatory cytokines, IL-1ß, IL-6 and TNFα, in response to TLR8 agonists. Thus, the presence of this variant confers a significant protective effect on disease severity. These results show for the first time the association between the M1V variant of TLR8 and reduced disease severity in RA, which could have prognostic value for these patients.

A Truncated Variant of ASCC1, a Novel Inhibitor of NF-κB, Is Associated with Disease Severity in Patients with Rheumatoid Arthritis.

Loss of the regulatory mechanisms that avoid excessive or constitutive activation of NF-κB may be associated with chronic inflammatory disorders, including rheumatoid arthritis (RA). After massive sequencing of 158 regulators of the NF-κB pathway in RA patients, we focused on a scarcely known gene, ASCC1, and showed that it potently inhibits the expression of NF-κB target genes (TRAIL, TNF-α, cIAP-1, IL8) and blocks activation of a NF-κB-luciferase reporter construct in five different human cell lines. Therefore, ASCC1 may contribute to avoiding a pathologic activation of this transcription factor. A truncated variant of ASCC1 (p.S78*) was found in RA patients and control individuals. Functional in vitro studies revealed that truncation abrogated the NF-κB inhibition capacity of ASCC1. In contrast with full-length protein, truncated ASCC1 did not reduce the transcriptional activation of NF-κB and the secretion of TNF-α in response to inflammatory stimuli. We analyzed the clinical impact of p.S78* variant in 433 patients with RA and found that heterozygous carriers of this variant needed more disease-modifying antirheumatic drugs, and more patients with this genotype needed treatment with corticoids and biologic agents. Moreover, the truncated allele-carrier group had lower rates of remission compared with the full-length variant carriers. Overall, our findings show for the first time, to our knowledge, that ASCC1 inhibits NF-κB activation and that a truncated and inactive variant of ASCC1 is associated with a more severe disease, which could have clinical value for assessing the progression and prognosis of RA.

Influence of interleukin 10 promoter polymorphisms in polymyalgia rheumatica: disease susceptibility and functional consequences.

OBJECTIVES: To investigate the functional consequences of IL10 (-592C/A and -1082A/G) gene polymorphisms and their association with susceptibility to, and disease phenotype, in patients with polymyalgia rheumatica (PMR). METHODS: A total number of 168 with PMR and 124 age-matched controls were genotyped using allele-specific primers and restriction fragment length polymorphism analysis. The levels of circulating IL10 and the production of IL10 by PBMCs after in vitro stimulation were studied by Cytometric Bead Array. RESULTS: No significant differences were observed in genotype or allele frequency distribution between patients and controls. The clinical characteristics and prognosis of these patients were also unrelated to the presence of these polymorphisms. No significant differences between PMR patients with low ESR (<40 mm/hr) and classic PMR (>40 mm/hr) were found. Furthermore, we did not observe any influence of circulating IL10 with the intensity of the acute phase response. In both, PMR patients and age-matched controls, no differences in circulating IL10 levels or IL10 production were observed depending on the genotypes of the IL10 gene. CONCLUSIONS: These results do not support the impact of IL10 variants in susceptibility or clinical phenotype of PMR patients. In this aged population no functional association was found between IL10 gene variants and IL10 production.

Cytokine gene considerations in giant cell arteritis: IL10 promoter polymorphisms and a review of the literature.

Polymorphisms of cytokine genes have been investigated as susceptibility markers of giant cell arteritis (GCA). Here, we have reviewed the evidence to date and especially addressed the functional consequences of IL10 (-592C/A and -1082A/G) gene polymorphisms and their association with susceptibility to and disease phenotype in GCA. A total number of 71 patients with GCA and 124 age-matched controls were genotyped using allele-specific primers and restriction fragment length polymorphism analysis. As previous studies in GCA showed inconsistent results, a meta-analysis of the existing studies was also conducted by using both fixed and random-effects models. The levels of circulating IL10 and the production of IL10 by peripheral blood mononuclear cells after in vitro stimulation were studied by Cytometric Bead Array. Data showed no significant differences in genotype or allele frequency distribution between patients and controls. The clinical characteristics and prognosis of these patients were also unrelated to the presence of these polymorphisms. However, the meta-analysis found a significant association of IL10 -592C/A polymorphism with susceptibility to GCA (odds ratio 2.205 (95% confidence interval 1.074-4.524); p = 0.031). In both patients and age-matched controls, no differences in circulating IL10 levels or IL10 production were observed depending on the genotypes of the IL10 gene. In conclusion, although our cohort results do not support the impact of IL10 variants in susceptibility or clinical phenotype of GCA patients, the meta-analysis revealed a significant association of -592C/A polymorphism with susceptibility to GCA. In this population, no functional association was found between IL10 gene variants and IL10 production.

Phagocyte dysfunction in polymyalgia rheumatica and other age-related, chronic, inflammatory conditions.

This study was conducted to evaluate phagocyte function in patients with age-related chronic inflammatory conditions. It included 95 patients with PMR, 17 with GCA, 40 with EORA, and 25 age-matched HCs. Serum IL-8 was determined with a bead array. The chemotactic capacity, phagocytic ability, and oxidative burst activity of circulating leukocytes were determined with flow cytometry kits. Patients with active chronic inflammatory diseases showed a significant increase in circulating levels of IL-8 that remained elevated in patients with PMR or EORA, despite treatment. No correlation was found between circulating IL-8 and the migratory capacity of neutrophils. Neutrophils from patients with active EORA without stimulus and after fMLP stimuli showed a higher capacity to migrate than those of the HCs (P=0.033). The phagocytic activity of granulocytes in the patients with GCA was significantly higher than in the HCs and the patients with PMR or EORA (P<0.05). The percentage and MFI of phagocytes that produce ROIs when stimulated with Escherichia coli was significantly reduced in neutrophils and monocytes from the patients with age-restricted inflammatory conditions. We concluded that the effector functions of phagocytes, determined to be chemotaxis, phagocytosis, and oxidative burst, are deregulated in age-restricted inflammatory disorders and may have a pathogenic role.

Induction of proinflammatory cytokines in human lung epithelial cells during Rhodococcus equi infection.

Rhodococcus equi is an opportunistic human pathogen associated with immunosuppressed people. While the interaction of R. equi with macrophages has been comprehensively studied, little is known about its interactions with non-phagocytic cells. Here, we characterized the entry process of this bacterium into human lung epithelial cells. The invasion is inhibited by nocodazole and wortmannin, suggesting that the phosphatidylinositol 3-kinase pathway and microtubule cytoskeleton are important for invasion. Pre-incubation of R. equi with a rabbit anti-R. equi polyclonal antiserum resulted in a dramatic reduction in invasion. Also, the invasion process as studied by immunofluorescence and scanning electron microscopy indicates that R. equi make initial contact with the microvilli of the A549 cells, and at the structural level, the entry process was observed to occur via a zipper-like mechanism. Infected lung epithelial cells upregulate the expression of cytokines IL-8 and IL-6 upon infection. The production of these pro-inflammatory cytokines was significantly enhanced in culture supernatants from cells infected with non-mucoid plasmid-less strains when compared with cells infected with mucoid strains. These results demonstrate that human airway epithelial cells produce pro-inflammatory mediators against R. equi isolates.

Análisis del polimorfismo rs20541 (R130Q) del gen de la IL-13 en pacientes con enfermedades inflamatorias crónicas asociadas al envejecimiento / Analysis of the rs20541 (R130Q) polymorphism in the IL-13 gene in patients with elderly-associated chronic inflammatory diseases

Objetivo: Investigar si existe asociación del polimorfismo rs20541 (R130Q) del gen de la IL-13 con la susceptibilidad y la expresión clínica de pacientes con enfermedades inflamatorias crónicas asociadas al envejecimiento. Material y métodos: Se estudiaron 78 pacientes con arteritis de células gigantes (ACG), 174 con polimialgia reumática (PMR), 90 con artritis reumatoide de comienzo en el anciano (EORA), y 465 controles sanos de la misma zona geográfica. El polimorfismo rs20541 (R130Q) para IL-13 se evaluó mediante PCR-RFLP. Los niveles de IL-13 circulante se determinaron por ELISA. Resultados: En los pacientes con ACG se observó una mayor frecuencia del genotipo AA [2,349 (0,994- 5,554)], así como del alelo A [1,589 (1,085-2,328)] y de portadores de dicho alelo [1,656 (1,021-2,686)] (p < 0,05). No encontramos diferencias significativas entre los pacientes con PMR y EORA respecto al grupo control. Cuando comparamos las diferentes patologías entre sí, tampoco encontramos diferencias significativas entre ellas. En los pacientes con ACG las diferencias en el genotipo se asociaron con el pronóstico de la enfermedad. En pacientes con PMR, el genotipo AA se asoció con niveles más elevados de IL-13 circulante comparado con el GA. Sin embargo, esta asociación no se apreció para los controles o las otras enfermedades. Conclusiones: La ACG es más frecuente en individuos portadores del polimorfismo rs20541 (R130Q) del gen de la IL13. La utilidad de este gen para predecir el pronóstico en ACG debe ser confirmada en estudios con mayor número de pacientes (AU)

Objective: To investigate whether there is association between the rs20541 (R130Q) polymorphism in the IL-13 gene with disease susceptibility and clinical subsets in patients with elderly-associated inflammatory chronic diseases. Material and methods: 78 patients with giant cell arteritis (GCA), 174 with polymyalgia rheumatica (PMR), 90 elderly-onset rheumatoid arthritis (EORA), and 465 healthy controls from the same geographic area were studied. The rs20541 (R130Q) polymorphism in the IL-13 gene was evaluated by PCR-RFLP. Circulating levels of IL-13 were measured by ELISA. Results: A higher frequency of the AA genotype [2.349 (0.994-5.554)], as well as the allele A [1.589 (1.085- 2.328] and the A carriers [1.656 (1.021-2.686)] (p < 0.05) was observed in the GCA patients. No significant differences were observed in the PMR and EORA patients as compared with the healthy controls. Neither difference was observed among the different disease groups studied. In GCA patients, differences in the genotype were associated with a worse prognosis. In PMR patients, the AA genotype was associated with higher levels of serum IL-13 than the GA one. However, such an association was not detected for controls and the other disease groups. Conclusions: GCA is more frequent in carriers of the rs20541 (R130Q) polymorphism in the IL-13 gene. The utility of this polymorphism to predict the GCA prognosis must be confirmed in studies with a higher number of patients (AU)

Relevancia de TLR4 en la infección por virus de la hepatitis C / Relevance of TLR4 in HCV infection

Objetivo: Se evaluó la frecuencia de los polimorfismos Asp-299Gly y Thr-399Ile en el gen deTLR4 en sujetos infectados por VHC y coinfectados por VIH y VHC y se estudió la expresión y función de TLR4 en función de esos polimorfismos. Material y métodos: El estudio incluyó 53 pacientes con infección por VHC, de los que 28 eran coinfectados además por VIH, más 30 sujetos controles sanos. Los polimorfismos Asp-299Glyy Thr-399Ile se determinaron mediante PCR-RFLP, las poblaciones celulares y la expresión deTLR4 se estudiaron mediante citometría de flujo, mientras que las citocinas proinflamatoriasse midieron en suero mediante CBA. Resultados: Se encontró un descenso significativo de células T CD4+ en los pacientes coinfectados por VHC y VIH. No se demostró una mayor susceptibilidad a sufrir la infección para ninguno de los 2 polimorfismos analizados. TLR4 estaba disminuido en células B, mientras que aumentaba en células T y monocitos. Se comprobó un aumento significativo de las citocinas proinflamatorias que fue el doble en coinfectados que en los portadores de la infección aislada por VHC. Estos cambios no mostraron relación con el polimorfismo estudiado. Discusión: La expresión de TLR4 en células T y monocitos se encuentra aumentada en la infección por VIH y se acompaña de un aumento en suero de citocinas proinflamatorias. Estos hallazgos no guardan relación con los polimorfismos Asp-299Gly y Thr-399Ile de TLR4 (AU)

Objective: The presence of the Asp-299Gly and Thr-399Ile polymorphisms in the toll-like receptor 4 (TLR4) gene was studied in subjects with HCV infection and HCV+HIV coinfection. The expression and function of TLR4 as regards these polymorphisms is assessed. Material and methods: The study included 53 patients infected with HCV, among whom 27had coinfection HCV+HIV, and 30 healthy subjects. The polymorphisms were studied by PCR-RFLP. The number of lymphocyte subsets, as well as TLR4 expression, was determined by flow cytometry, and the concentration of cytokines was measured in serum by (cytometricbead assay) CBA. Results: CD4+T cells were significantly decreased in patients coinfected with HCV+HIV. There was no association between the presence of any of the two studied polymorphisms and the susceptibility to suffer from infection. TLR4 was less expressed in B cells, whereas it was increased in T cells and, in particular, monocytes. A significant increase in the levels of circulating pro-inflammatory cytokines was found, being two-fold increased incoinfected subjects as compared with patients with isolated HCV infection. None of the findings in cell subsets, TLR4 expression and cytokines was associated with the studied TLR4polymorphism.Discussion: The expression of TLR4 in T cells and monocytes is increased in HCV infection and is accompanied by increased serum levels of proinflammatory cytokines. These findings do not have any relationship with the Asp-299Gly and Thr-399Ile polymorphism (AU)

Analysis of the rs20541 (R130Q) polymorphism in the IL-13 gene in patients with elderly-associated chronic inflammatory diseases.

OBJECTIVE: To investigate whether there is association between the rs20541 (R130Q) polymorphism in the IL-13 gene with disease susceptibility and clinical subsets in patients with elderly-associated inflammatory chronic diseases. MATERIAL AND METHODS: 78 patients with giant cell arteritis (GCA), 174 with polymyalgia rheumatica (PMR), 90 elderly-onset rheumatoid arthritis (EORA), and 465 healthy controls from the same geographic area were studied. The rs20541 (R130Q) polymorphism in the IL-13 gene was evaluated by PCR-RFLP. Circulating levels of IL-13 were measured by ELISA. RESULTS: A higher frequency of the AA genotype [2.349 (0.994-5.554)], as well as the allele A [1.589 (1.085-2.328] and the A carriers [1.656 (1.021-2.686)] (p<0.05) was observed in the GCA patients. No significant differences were observed in the PMR and EORA patients as compared with the healthy controls. Neither difference was observed among the different disease groups studied. In GCA patients, differences in the genotype were associated with a worse prognosis. In PMR patients, the AA genotype was associated with higher levels of serum IL-13 than the GA one. However, such an association was not detected for controls and the other disease groups. CONCLUSIONS: GCA is more frequent in carriers of the rs20541 (R130Q) polymorphism in the IL-13 gene. The utility of this polymorphism to predict the GCA prognosis must be confirmed in studies with a higher number of patients.

Aging is associated with circulating cytokine dysregulation.

PURPOSE: Aging is accompanied by a progressive increase in pro-inflammatory cytokine status. However, little is known about the development of age-dependent modifications in other circulating cytokines. The aim of this study was to investigate in vivo the influence of age on circulating cytokine production in healthy subjects (HC). METHODS: Circulating cytokines were measured by CBA and ELISA in 73 HC. Intracellular cytokine production was assessed in CD3+ and CD14+ cells by flow cytometry. Production of cytokines in cell culture supernatants was also studied after polyclonal stimulation. RESULTS: Subjects were divided into three different groups according to age: 28 young HC (<30 years, 26.2 ± 2.4), 24 middle age HC (30-60 years, 44.7 ± 8.4) and 21 elderly HC (>60 years, 70.6 ± 7.9). Age was positively correlated with the circulating levels of IL-12p70, IL-1ß, TNFα, IL-6, and IL-10. Age had a negative correlation with circulating levels of IL-17. Besides, age was positively correlated with spontaneous intracellular expression of proinflammatory cytokines in circulating monocytes. No correlation was found with other intracellular cytokine expression or with the production of cytokines in cell culture supernatants after in vitro stimulation. Gender had a marginal effect on the circulating cytokine profile. CONCLUSION: Aging has a significant impact on the production of circulating cytokines in healthy individuals. The circulating cytokine milieu may contribute to the development of age-restricted conditions.