9 research outputs found
Calcineurin inhibitors cyclosporine A and tacrolimus induce vascular inflammation and endothelial activation through TLR4 signaling
The introduction of the calcineurin inhibitors (CNIs) cyclosporine and tacrolimus greatly reduced the
rate of allograft rejection, although their chronic use is marred by a range of side effects, among them
vascular toxicity. In transplant patients, it is proved that innate immunity promotes vascular injury
triggered by ischemia-reperfusion damage, atherosclerosis and hypertension. We hypothesized that
activation of the innate immunity and inflammation may contribute to CNI toxicity, therefore we
investigated whether TLR4 mediates toxic responses of CNIs in the vasculature. Cyclosporine and
tacrolimus increased the production of proinflammatory cytokines and endothelial activation markers
in cultured murine endothelial and vascular smooth muscle cells as well as in ex vivo cultures of murine
aortas. CNI-induced proinflammatory events were prevented by pharmacological inhibition of TLR4.
Moreover, CNIs were unable to induce inflammation and endothelial activation in aortas from TLR4−/−
mice. CNI-induced cytokine and adhesion molecules synthesis in endothelial cells occurred even in the
absence of calcineurin, although its expression was required for maximal effect through upregulation
of TLR4 signaling. CNI-induced TLR4 activity increased O2
−/ROS production and NF-κB-regulated
synthesis of proinflammatory factors in cultured as well as aortic endothelial and VSMCs. These data
provide new insight into the mechanisms associated with CNI vascular inflammationThis work was supported by grants from the Instituto de Salud Carlos III (Ministerio de Economía Competitividad, Gobierno de España): FEDER funds ISCIII RETIC REDINREN RD12/0021, PI11/02242, PI13/00047, PI14/0041, PI14/00386, PI15/01460; Comunidad de Madrid (CIFRA S2010/BMD-2378); Sociedad Española de Nefrología. Salary support: RR-D:
CIFRA; CO-S: Fundación Conchita Rábago de Jiménez Díaz; CG-G and RRR-D: REDINREN; AO: Programa Intensificación Actividad Investigadora (ISCIII/Agencia Laín-Entralgo/CM); JE and MRO: Universidad Autónoma de Madrid; AMR: Contrato Miguel Serve (ISCIII
Micro-arrayed wheat seed and grass pollen allergens for component-resolved diagnosis.
Background: Wheat is a potent allergen source and can cause baker's asthma, food and pollen allergy. The aim of the study was to develop an allergen micro-array for differential diagnosis of baker's asthma, wheat-induced food allergy and grass pollen allergy. Methods: We analysed the immunoglobulin-E reactivity profiles of patients suffering from baker's asthma, wheat-induced food allergy and grass pollen allergy to micro-arrayed recombinant wheat flour allergens and grass pollen allergens and compared these results with clinical results and diagnostic tests based on crude wheat flour, wheat pollen and grass pollen allergen extracts. Results: We identified recombinant wheat flour allergens, which are specifically recognized by patients suffering from baker's asthma, but not from patients with food allergy to wheat or pollen allergy. rPhl p 1 and rPhl p 5 were identified as marker allergens specific for grass pollen allergy. They can be used to replace grass pollen extracts for allergy diagnosis and to identify grass pollen allergic patients among patients suffering from baker's asthma and wheat-induced food allergy. Profilin was identified as a cross-reactive allergen recognized by patients suffering from baker's asthma, food and pollen allergy. Conclusions: Our results indicate that it will be possible to design serological tests based on micro-arrayed recombinant wheat seed and grass pollen allergens for the discrimination of baker's asthma, wheat-induced food allergy and grass pollen allergy
Interleukin-17A blockade reduces albuminuria and kidney injury in an accelerated model of diabetic nephropathy
Diabetic nephropathy (DN) is one of the most common
complications of diabetes, and currently the first end-stage
renal disease worldwide. New strategies to treat DN using
agents that target inflammatory pathways have attracted
special interest. Recent pieces of evidences suggest a
promising effect of IL-17A, the Th17 effector cytokine.
Among experimental DN models, mouse strain BTBR ob/ob
(leptin deficiency mutation) develops histological features
similar to human DN, which means an opportunity to study
mechanisms and novel therapies aimed at DN regression.
We found that BTBR ob/ob mice presented renal activation
of the factors controlling Th17 differentiation. The presence
of IL-17A-expressing cells, mainly CD4D and gd
lymphocytes, was associated with upregulation of
proinflammatory factors, macrophage infiltration and the
beginning of renal damage. To study IL-17A involvement in
experimental DN pathogenesis, treatment with an IL-17A
neutralizing antibody was carried out starting when the
renal damage had already appeared. IL-17A blockade
ameliorated renal dysfunction and disease progression in
BTBR ob/ob mice. These beneficial effects correlated to
podocyte number restoration and inhibition of NF-kB/
proinflammatory factors linked to a decrease in renal
inflammatory-cell infiltration. These data demonstrate that
IL-17A takes part in diabetes-mediated renal damage and
could be a promising therapeutic target to improve DN.This work was supported by grants PAI 82140017 to CL; Fondecyt
1160465 to SM; Division of Nephrology, Universidad Austral de Chile,
the Instituto de Salud Carlos III and FEDER European Union funds
(PI14/00041, PI17/00119 to MR-O, and PI14/00386 and PI17/01495 to
JE); Red de Investigación Renal (REDinREN; RD16/009) and
Comunidad de Madrid (B2017/BMD-3751 NOVELREN-CM) to MR-O,
and Sociedad Española de Nefrologí
Molecular and immunological characterization of a wheat serine proteinase inhibitor as a novel allergen in baker's asthma
IgE-mediated sensitization to wheat flour belongs to the most frequent causes of occupational asthma. A cDNA library from wheat seeds was constructed and screened with serum IgE from baker's asthma patients. One IgE-reactive phage clone contained a full-length cDNA coding for an allergen with a molecular mass of 9.9 kDa and an isoelectric point of 6. According to sequence analysis it represents a member of the potato inhibitor I family, a group of serine proteinase inhibitors, and thus is the first allergen belonging to the group 6 pathogenesis-related proteins. The recombinant wheat seed proteinase inhibitor was expressed in Escherichia coli and purified to homogeneity. According to circular dichroism analysis, it represented a soluble and folded protein with high thermal stability containing mainly beta-sheets, random coils, and an alpha-helical element. The recombinant allergen showed allergenic activity in basophil histamine release assays and reacted specifically with IgE from 3 of 22 baker's asthma patients, but not with IgE from grass pollen allergic patients or patients suffering from food allergy to wheat. Allergen-specific Abs were raised to localize the allergen by immunogold electron microscopy in the starchy endosperm and the aleuron layer. The allergen is mainly expressed in mature wheat seeds and, despite an approximately 50% sequence identity, showed no relevant cross-reactivity with allergens from other plant-derived food sources such as maize, rice, beans, or potatoes. Recombinant wheat serine proteinase inhibitor, when used in combination with other specific allergens, may be useful for the diagnosis and therapy of IgE-mediated baker's asthma
Author Correction: Combination of KIR2DS4 and FcγRIIa polymorphisms predicts the response to cetuximab in KRAS mutant metastatic colorectal cancer (Scientific Reports, (2019), 9, 1, (2589), 10.1038/s41598-019-39291-2)
The original version of this Article contained an error in the spelling of the author P. Garcia-Alfonso, which was incorrectly given as P. Garcia. This error has now been corrected in the PDF and HTML versions of this Article
Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19).
The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situation
Characteristics and predictors of death among 4035 consecutively hospitalized patients with COVID-19 in Spain.
To analyse the characteristics and predictors of death in hospitalized patients with coronavirus disease 2019 (COVID-19) in Spain. A retrospective observational study was performed of the first consecutive patients hospitalized with COVID-19 confirmed by real-time PCR assay in 127 Spanish centres until 17 March 2020. The follow-up censoring date was 17 April 2020. We collected demographic, clinical, laboratory, treatment and complications data. The primary endpoint was all-cause mortality. Univariable and multivariable Cox regression analyses were performed to identify factors associated with death. Of the 4035 patients, male subjects accounted for 2433 (61.0%) of 3987, the median age was 70 years and 2539 (73.8%) of 3439 had one or more comorbidity. The most common symptoms were a history of fever, cough, malaise and dyspnoea. During hospitalization, 1255 (31.5%) of 3979 patients developed acute respiratory distress syndrome, 736 (18.5%) of 3988 were admitted to intensive care units and 619 (15.5%) of 3992 underwent mechanical ventilation. Virus- or host-targeted medications included lopinavir/ritonavir (2820/4005, 70.4%), hydroxychloroquine (2618/3995, 65.5%), interferon beta (1153/3950, 29.2%), corticosteroids (1109/3965, 28.0%) and tocilizumab (373/3951, 9.4%). Overall, 1131 (28%) of 4035 patients died. Mortality increased with age (85.6% occurring in older than 65 years). Seventeen factors were independently associated with an increased hazard of death, the strongest among them including advanced age, liver cirrhosis, low age-adjusted oxygen saturation, higher concentrations of C-reactive protein and lower estimated glomerular filtration rate. Our findings provide comprehensive information about characteristics and complications of severe COVID-19, and may help clinicians identify patients at a higher risk of death