Influencia de marcadores genéticos del hospedador en la historia natural de la infección por el virus de la Hepatitis C

La infección por el virus de la hepatitis C (VHC) constituye un serio problema de salud pública debido a su elevada prevalencia, ya que afecta a más de 180 millones de personas en el mundo. Tras la infección aguda, alrededor de un 25% de los pacientes eliminan el virus espontáneamente, proceso denominado aclaramiento viral espontáneo (AVE), pero en el resto de los pacientes la viremia persiste cronificándose la infección. Posteriormente, los pacientes evolucionan de manera muy diferente y presentan respuestas diversas al mismo tratamiento aún en el caso de condiciones homogéneas de infección. Por tanto, deben existir factores genéticos del hospedador que condicionen estas diferentes respuestas. Uno de los objetivos de numerosos grupos de investigación es determinar qué genes del hospedador pueden estar implicados en la historia natural de la infección por el VHC así como de replicar en distintas poblaciones los marcadores genéticos ya identificados para poder validarlos, y así podamos predecir el curso clínico de la infección y optimizar el uso de los diversos agentes terapéuticos disponibles. El objetivo principal de este trabajo ha sido replicar en nuestra población la asociación descrita entre el aclaramiento viral espontáneo o la respuesta al tratamiento en la infección por el virus de hepatitis C y los genes IL28B, HAVCR1, TANK, TNFSF18 e IL18BP. Así, el diseño experimental utilizado consistió en un estudio caso-control analizando varios polimorfismos en estos genes candidatos. El primer gen seleccionado, la interleuquina 28B (IL28B), se incluyó en este estudio tras publicarse en tres GWAS una asociación entre el locus IL28B y la respuesta a la terapia combinada con IFN-¿ y ribavirina (RBV) en pacientes infectados por el VHC de genotipo 1 (G1) (Ge et al, 2009; Suppiah et al, 2009; Tanaka et al, 2009). Poco después de la identificación de IL28B, los otros 4 genes incluidos en este trabajo, fueron identificados, de entre 112 genes seleccionados por su función inmunológica, por su asociación con la resolución espontánea del VHC en un estudio con pacientes americanos de origen europeo o africano (Mosbruger et al, 2010). Para estudiar la influencia o relación de estos 5 genes con la infección por el VHC, se utilizó una cohorte de pacientes españoles monoinfectados con el VHC, de los que teníamos datos de respuesta al tratamiento y del grado de fibrosis, y otra de pacientes que aclararon el VHC de manera espontánea. Debido a la relación entre la respuesta al tratamiento y el genotipo viral que causa la infección, tuvimos en cuenta en nuestros análisis el genotipo viral de los pacientes, e incluimos una tercera cohorte de coinfectados con VIH (VHC/VIH) que aportaba un mayor número de pacientes infectados por genotipo viral no 1 (No-G1) para confirmar determinadas asociaciones encontradas. En nuestro estudio en población española se confirmó que el genotipo rs12979860CC estaba asociado con una mejor respuesta al tratamiento, tanto en la cohorte de pacientes monoinfectados (60,2% vs. 39,8%) como en la cohorte de coinfectados (66,2% vs. 33,8%). Los pacientes con genotipo rs12979860CC respondían mejor al tratamiento independientemente de que el tratamiento recibido hubiese sido monoterapia (60,3% vs. 39,7%) o terapia combinada (60% vs. 40%). Por otra parte, los pacientes con genotipo rs12979860CC eran mayoritariamente infectados por No-G1 en comparación con G1 (66,7% vs 39,1%). Nuestros resultados demostraron también que el genotipo rs12979860CC era mucho más frecuente en los individuos con aclaramiento viral espontáneo que en los individuos VHC crónicos (72,5% vs 45,6%). Con respecto al gen HAVCR1, demostramos que había una relación entre este gen y el genotipo viral. Entre los 4 haplotipos (A-D) descritos en este gen, los pacientes con haplotipo C tenían la mayor tasa de infección por virus G1 (75,82%), por el contrario los individuos con haplotipo D tenían la menor tasa de infección por virus G1 (57,72%), mientras que los individuos con haplotipo A o B tenían valores intermedios (66%). Con respecto al gen TANK, se encontró asociación con el grado de fibrosis en las dos cohortes de pacientes estudiadas, monoinfectados y coinfectados, mientras que los genes TNFSF18 e IL18BP no se asociaron con ninguna de las variables estudiadas. En conclusión, nuestros resultados confirman que factores genéticos del hospedador influyen en el curso natural de la infección, y tendrían implicaciones en el manejo terapéutico de los pacientes.Premio Extraordinario de Doctorado U

The age again in the eye of the COVID-19 storm: evidence-based decision making

Coronavirus SARS-CoV-2; COVID-19; 2019-nCoV; Immunosenescència; ConfinamentCoronavirus SARS-CoV-2; COVID-19; 2019-nCoV; Inmunosenescencia; ConfinamientoCoronavirus SARS-CoV-2; COVID-19; 2019-nCoV; Immunosenescence; LockdownBackground One hundred fifty million contagions, more than 3 million deaths and little more than 1 year of COVID-19 have changed our lives and our health management systems forever. Ageing is known to be one of the significant determinants for COVID-19 severity. Two main reasons underlie this: immunosenescence and age correlation with main COVID-19 comorbidities such as hypertension or dyslipidaemia. This study has two aims. The first is to obtain cut-off points for laboratory parameters that can help us in clinical decision-making. The second one is to analyse the effect of pandemic lockdown on epidemiological, clinical, and laboratory parameters concerning the severity of the COVID-19. For these purposes, 257 of SARSCoV2 inpatients during pandemic confinement were included in this study. Moreover, 584 case records from a previously analysed series, were compared with the present study data. Results Concerning the characteristics of lockdown series, mild cases accounted for 14.4, 54.1% were moderate and 31.5%, severe. There were 32.5% of home contagions, 26.3% community transmissions, 22.5% nursing home contagions, and 8.8% corresponding to frontline worker contagions regarding epidemiological features. Age > 60 and male sex are hereby confirmed as severity determinants. Equally, higher severity was significantly associated with higher IL6, CRP, ferritin, LDH, and leukocyte counts, and a lower percentage of lymphocyte, CD4 and CD8 count. Comparing this cohort with a previous 584-cases series, mild cases were less than those analysed in the first moment of the pandemic and dyslipidaemia became more frequent than before. IL-6, CRP and LDH values above 69 pg/mL, 97 mg/L and 328 U/L respectively, as well as a CD4 T-cell count below 535 cells/μL, were the best cut-offs predicting severity since these parameters offered reliable areas under the curve. Conclusion Age and sex together with selected laboratory parameters on admission can help us predict COVID-19 severity and, therefore, make clinical and resource management decisions. Demographic features associated with lockdown might affect the homogeneity of the data and the robustness of the results.This work has been carried out without funding

PKM2 subcellular localization is involved in oxaliplatin resistance acquisition in HT29 human colorectal cancer cell lines

Ajuts: Beca bianual de la Fundació Olga Torres 2008-2009Chemoresistance is the main cause of treatment failure in advanced colorectal cancer (CRC). However, molecular mechanisms underlying this phenomenon remain to be elucidated. In a previous work we identified low levels of PKM2 as a putative oxaliplatin-resistance marker in HT29 CRC cell lines and also in patients. In order to assess how PKM2 influences oxaliplatin response in CRC cells, we silenced PKM2 using specific siRNAs in HT29, SW480 and HCT116 cells. MTT test demonstrated that PKM2 silencing induced resistance in HT29 and SW480 cells and sensitivity in HCT116 cells. Same experiments in isogenic HCT116 p53 null cells and double silencing of p53 and PKM2 in HT29 cells failed to show an influence of p53. By using trypan blue stain and FITC-Annexin V/PI tests we detected that PKM2 knockdown was associated with an increase in cell viability but not with a decrease in apoptosis activation in HT29 cells. Fluorescence microscopy revealed PKM2 nuclear translocation in response to oxaliplatin in HCT116 and HT29 cells but not in OXA-resistant HTOXAR3 cells. Finally, by using a qPCR Array we demonstrated that oxaliplatin and PKM2 silencing altered cell death gene expression patterns including those of BMF, which was significantly increased in HT29 cells in response to oxaliplatin, in a dose and time-dependent manner, but not in siPKM2-HT29 and HTOXAR3 cells. BMF gene silencing in HT29 cells lead to a decrease in oxaliplatin-induced cell death. In conclusion, our data report new non-glycolytic roles of PKM2 in response to genotoxic damage and proposes BMF as a possible target gene of PKM2 to be involved in oxaliplatin response and resistance in CRC cells

Case Report: Successful Lung Transplantation from a Donor Seropositive for Trypanosoma cruzi Infection (Chagas Disease) to a Seronegative Recipient

Lung transplantation; Seropositive donor; Trypanosoma cruziTrasplantament de pulmó; Donant seropositiu; Trypanosoma cruziTrasplante de pulmón; Donante seropositivo; Trypanosoma cruziThe increasing shortage of organs for transplantation has prompted transplant programs to investigate the use of extended criteria donors, such as those with transmissible infectious diseases. Successful cases of organ transplantation (mostly kidney and liver) from Trypanosoma cruzi seropositive donors to seronegative recipients have been reported. We present a case of lung transplantation from a donor serologically positive for Chagas disease to a seronegative recipient, and provide a review of the literature. Left single lung transplantation was performed in a 44-year-old Spanish woman presenting with interstitial lung disease in February 2016. The deceased donor was a Colombian immigrant living in Spain who was serologically positive for Chagas disease. Oral administration of 5 mg/kg/day benznidazole divided in three doses for 60 days was given for specific Chagas disease prophylaxis after transplantation. Periodic follow-up with serological reverse transcription polymerase chain reaction to detect T. cruzi DNA were performed until 6 months after the end of treatment. All results were negative, indicating that transmission of T. cruzi had not occurred. In a review of the literature, two similar cases were identified in Argentina and the United States. In both cases T. cruzi infection was detected posttransplant in the recipients, after which they were treated with benznidazole. The course of the patient described herein confirms that lungs from donors with chronic T. cruzi infection can be used successfully as allografts, and that posttransplant prophylaxis with benznidazole may reduce the probability of transmission of T. cruzi to the recipient

Allele and haplotype frequencies of HLA-A, -B, -C, -DRB1, -DQB1 and -DQA1 in Castile and Leon region from North West of Spain

HLA studies have been used to determine the admixture of different populations within the Iberian Peninsula including neighbouring regions with shared origins, such as Portugal and Castile and Leon. These studies certainly can be used to study human migration that could establish populations currently settled according to genetic distant analysis based on the HLA diversity and language variety.This work was supported by the “Gerencia Regional de Salud de Castilla y Leon” (GRS 2080/A/19, 2019) and (GRS COVID 70/A/20, 2020)

Free PCR virus detection via few-layer bismuthene and tetrahedral DNA nanostructured assemblies

In this work we describe a highly sensitive method based on a biocatalyzed electrochemiluminescence approach. The system combines, for the first time, the use of few-layer bismuthene (FLB) as a platform for the oriented immobilization of tetrahedral DNA nanostructures (TDNs) specifically designed and synthetized to detect a specific SARS-CoV-2 gene sequence. In one of its vertices, these TDNs contain a DNA capture probe of the open reading frame 1 ab (ORF1ab) of the virus, available for the biorecognition of the target DNA/RNA. At the other three vertices, there are thiol groups that enable the stable anchoring/binding to the FLB surface. This novel geometry/approach enables not only the binding of the TDNs to surfaces, but also the orientation of the capture probe in a direction normal to the bismuthine surface so that it is readily accessible for binding/recognition of the specific SARS-CoV-2 sequence. The analytical signal is based on the anodic electrochemiluminescence (ECL) intensity of luminol which, in turn, arises as a result of the reaction with H2O2, generated by the enzymatic reaction of glucose oxidation, catalyzed by the biocatalytic label avidin-glucose oxidase conjugate (Av-GOx), which acts as co-reactant in the electrochemiluminescent reaction. The method exhibits a limit of detection (LOD) of 4.31 aM and a wide linear range from 14.4 aM to 1.00 μM, and its applicability was confirmed by detecting SARS-CoV-2 in nasopharyngeal samples from COVID-19 patients without the need of any amplification processPID2020-116728RB-I00, PID2020-116661RB-I00, PID2020-119352RB-I00, PDC2021-120782-C2, PID2022-138908NB-C31, CTQ2015-71955-REDT, S2018/NMT-434

The age again in the eye of the COVID-19 storm: evidence-based decision making

Background: One hundred fifty million contagions, more than 3 million deaths and little more than 1 year of COVID-19 have changed our lives and our health management systems forever. Ageing is known to be one of the significant determinants for COVID-19 severity. Two main reasons underlie this: immunosenescence and age correlation with main COVID-19 comorbidities such as hypertension or dyslipidaemia. This study has two aims. The first is to obtain cut-off points for laboratory parameters that can help us in clinical decision-making. The second one is to analyse the effect of pandemic lockdown on epidemiological, clinical, and laboratory parameters concerning the severity of the COVID-19. For these purposes, 257 of SARSCoV2 inpatients during pandemic confinement were included in this study. Moreover, 584 case records from a previously analysed series, were compared with the present study data. Results: Concerning the characteristics of lockdown series, mild cases accounted for 14.4, 54.1% were moderate and 31.5%, severe. There were 32.5% of home contagions, 26.3% community transmissions, 22.5% nursing home contagions, and 8.8% corresponding to frontline worker contagions regarding epidemiological features. Age > 60 and male sex are hereby confirmed as severity determinants. Equally, higher severity was significantly associated with higher IL6, CRP, ferritin, LDH, and leukocyte counts, and a lower percentage of lymphocyte, CD4 and CD8 count. Comparing this cohort with a previous 584-cases series, mild cases were less than those analysed in the first moment of the pandemic and dyslipidaemia became more frequent than before. IL-6, CRP and LDH values above 69 pg/mL, 97 mg/L and 328 U/L respectively, as well as a CD4 T-cell count below 535 cells/?L, were the best cut-offs predicting severity since these parameters offered reliable areas under the curve. Conclusion: Age and sex together with selected laboratory parameters on admission can help us predict COVID-19 severity and, therefore, make clinical and resource management decisions. Demographic features associated with lockdown might affect the homogeneity of the data and the robustness of the results

COVID-19 : Age, Interleukin-6, C-reactive protein, and lymphocytes as key clues from a multicentre retrospective study

Background: The SARS-CoV-2 infection has widely spread to become the greatest public health challenge to date, the COVID-19 pandemic. Different fatality rates among countries are probably due to non-standardized records being carried out by local health authorities. The Spanish case-fatality rate is 11.22%, far higher than those reported in Asia or by other European countries. A multicentre retrospective study of demographic, clinical, laboratory and immunological features of 584 Spanish COVID-19 hospitalized patients and their outcomes was performed. The use of renin-angiotensin system blockers was also analysed as a risk factor. Results: In this study, 27.4% of cases presented a mild course, 42.1% a moderate one and for 30.5% of cases, the course was severe. Ages ranged from 18 to 98 (average 63). Almost 60 % (59.8%) of patients were male. Interleukin 6 was higher as severity increased. On the other hand, CD8 lymphocyte count was significantly lower as severity grew and subpopulations CD4, CD8, CD19, and NK showed concordant lowering trends. Severity-related natural killer percent descents were evidenced just within aged cases. A significant severity-related decrease of CD4 lymphocytes was found in males. The use of angiotensin-converting enzyme inhibitors was associated with a better prognosis. The angiotensin II receptor blocker use was associated with a more severe course. Conclusions: Age and age-related comorbidities, such as dyslipidaemia, hypertension or diabetes, determined more frequent severe forms of the disease in this study than in previous literature cohorts. Our cases are older than those so far reported and the clinical course of the disease is found to be impaired by age. Immunosenescence might be therefore a suitable explanation for the hampering of immune system effectors. The adaptive immunity would become exhausted and a strong but ineffective and almost deleterious innate response would account for COVID-19 severity. Angiotensin-converting enzyme inhibitors used by hypertensive patients have a protective effect in regards to COVID-19 severity in our series. Conversely, patients on angiotensin II receptor blockers showed a severer disease

Endothelial dysfunction is an early indicator of sepsis and neutrophil degranulation of septic shock in surgical patients

Producción CientíficaBackground: Stratification of the severity of infection is currently based on the Sequential Organ Failure Assessment (SOFA) score, which is difficult to calculate outside the ICU. Biomarkers could help to stratify the severity of infection in surgical patients. Methods: Levels of ten biomarkers indicating endothelial dysfunction, 22 indicating emergency granulopoiesis, and six denoting neutrophil degranulation were compared in three groups of patients in the first 12 h after diagnosis at three Spanish hospitals. Results: There were 100 patients with infection, 95 with sepsis and 57 with septic shock. Seven biomarkers indicating endothelial dysfunction (mid-regional proadrenomedullin (MR-ProADM), syndecan 1, thrombomodulin, angiopoietin 2, endothelial cell-specific molecule 1, vascular cell adhesion molecule 1 and E-selectin) had stronger associations with sepsis than infection alone. MR-ProADM had the highest odds ratio (OR) in multivariable analysis (OR 11·53, 95 per cent c.i. 4·15 to 32·08; P = 0·006) and the best area under the curve (AUC) for detecting sepsis (0·86, 95 per cent c.i. 0·80 to 0·91; P < 0·001). In a comparison of sepsis with septic shock, two biomarkers of neutrophil degranulation, proteinase 3 (OR 8·09, 1·34 to 48·91; P = 0·028) and lipocalin 2 (OR 6·62, 2·47 to 17·77; P = 0·002), had the strongest association with septic shock, but lipocalin 2 exhibited the highest AUC (0·81, 0·73 to 0·90; P < 0·001). Conclusion: MR-ProADM and lipocalin 2 could be alternatives to the SOFA score in the detection of sepsis and septic shock respectively in surgical patients with infection.Instituto de Salud Carlos III (grants PI15/01959, PI15/01451 and PI16/01156