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Background: Approximately 13.8% and 6.1% of coronavirus disease 2019 (COVID-19) patients require hospitalization and sometimes intensive care unit (ICU) admission, respectively. There is no biomarker to predict which of these patients will develop an aggressive stage that we could improve their quality of life and healthcare management. Our main goal is to include new markers for the classification of COVID-19 patients. Methods: Two tubes of peripheral blood were collected from a total of 66 (n = 34 mild and n = 32 severe) samples (mean age 52 years). Cytometry analysis was performed using a 15-parameter panel included in the Maxpar® Human Monocyte/Macrophage Phenotyping Panel Kit. Cytometry by time-of-flight mass spectrometry (CyTOF) panel was performed in combination with genetic analysis using TaqMan® probes for ACE2 (rs2285666), MX1 (rs469390), and TMPRSS2 (rs2070788) variants. GemStone™ and OMIQ software were used for cytometry analysis. Results: The frequency of CD163+/CD206- population of transitional monocytes (T-Mo) was decreased in the mild group compared to that of the severe one, while T-Mo CD163-/CD206- were increased in the mild group compared to that of the severe one. In addition, we also found differences in CD11b expression in CD14dim monocytes in the severe group, with decreased levels in the female group (p = 0.0412). When comparing mild and severe disease, we also found that CD45- [p = 0.014; odds ratio (OR) = 0.286, 95% CI 0.104–0.787] and CD14dim/CD33+ (p = 0.014; OR = 0.286, 95% CI 0.104–0.787) monocytes were the best options as biomarkers to discriminate between these patient groups. CD33 was also indicated as a good biomarker for patient stratification by the analysis of GemStone™ software. Among genetic markers, we found that G carriers of TMPRSS2 (rs2070788) have an increased risk (p = 0.02; OR = 3.37, 95% CI 1.18–9.60) of severe COVID-19 compared to those with A/A genotype. This strength is further increased when combined with CD45-, T-Mo CD163+/CD206-, and C14dim/CD33+. Conclusions: Here, we report the interesting role of TMPRSS2, CD45-, CD163/CD206, and CD33 in COVID-19 aggressiveness. This strength is reinforced for aggressiveness biomarkers when TMPRSS2 and CD45-, TMPRSS2 and CD163/CD206, and TMPRSS2 and CD14dim/CD33+ are combined., Desarrollo e Innovación (I+D+i) en Biomedicina y en Ciencias de la Salud en Andalucía, ́ FEDER”, internal code PECOVID-0006-2020, by Consejería de ́ Salud, Junta de Andalucía, Uso de la metodologíá NGS y CYTOF para caracterizar a los pacientes con COVID-19”, internal code CV20-36740, by Secretaria General Universidades, Investigación y Tecnología, Junta de Andalucía

En la presente Tesis doctoral, hemos evaluado el impacto que la administración de MenSCs tiene sobre dos modelos murinos de inflamación aguda: el modelo de peritonitis inducida por tioglicolato (TGC) y el modelo de sepsis inducida por la administración de distintas cepas de Salmonella Typhimurium. En el modelo de peritonitis, la inyección intraperitoneal de MenSCs alteró el patrón de reclutamiento local de macrófagos, neutrófilos y otras poblaciones inmunológicas. Además, el número y porcentaje de las poblaciones peritoneales presentes era dependiente del momento de administración de las MenSCs. Las MenSCs administradas en el momento de reclutamiento de macrófagos tras la inyección de TGC, fueron capaces de migrar a órganos distantes y, cerca de la zona de inyección, promovieron la generación de agregados peritoneales compuestos por las propias células, macrófagos y neutrófilos. En ensayos in vitro, la estimulación de las MenSCs con TGC alteró su perfil de expresión, potenciando la transcripción de mediadores quimioatrayentes y antiinflamatorios. En el modelo de sepsis, por otro lado, la administración de MenSCs redujo el reclutamiento de macrófagos y neutrófilos al foco de infección, afectando a sus capacidades microbicidas y favoreciendo la diseminación bacteriana hacia otros órganos. Mediante ensayos in vitro de cocultivo con macrófagos humanos, confirmamos que las MenSCs alteraban el perfil fenotípico de estas células y afectaban a sus propiedades bactericidas. A pesar de incrementar la capacidad fagocítica de los macrófagos, el cocultivo con MenSCs redujo el grado de eliminación bacteriana y la producción de especies reactivos de oxígeno. En conclusión, estos hallazgos ponen de manifiesto cómo las MenSCs mantienen un «diálogo» con el microambiente que las rodea, modulando así propiedades funcionales y fenotípicas de las poblaciones inmunológicas innatas, principalmente macrófagos. Dada la enorme influencia que los macrófagos poseen sobre el contexto inflamatorio, es indispensable tener en cuenta esta interacción entre macrófagos y MenSCs si queremos asegurar la eficacia terapéutica de estas últimas en un entorno clínico. Por otro lado, los modelos experimentales descritos en esta Tesis son herramientas útiles para analizar los efectos moduladores de las MenSCs, y otras fuentes de MSCs, en patologías severas específicas., In this dissertation, we assessed the impact that MenSCs administration had in two mouse models of acute inflammation: a thioglycollate (TGC)-elicited peritonitis model and the Salmonella Typhimurium sepsis model. We found that in the TGC model, MenSCs intraperitoneal injection altered the local recruitment of macrophages, neutrophils and other innate immune cells. Moreover, this variation in the number and percentage of peritoneal populations was dependent on MenSCs administration timepoint. MenSCs administered at the onset of macrophage recruitment migrated to distant organs and promoted, locally, the formation of peritoneal aggregates. These aggregates harboured macrophages, neutrophils and the injected MenSCs. In vitro, the stimulation of MenSCs with TGC altered their expression profile enhancing the transcription of both chemoattracting and antiinflammatory mediators. In the Salmonella sepsis model, MenSCs exacerbated infection by diminishing the recruitment of macrophages and neutrophils to the site of injection and prevented bacterial clearance. Additional in vitro studies confirmed that co-culture of MenSCs with human macrophages reshaped their phenotype and impaired their bactericidal properties, affecting macrophage bacterial killing and the production of reactive oxygen intermediates. In conclusion, our findings highlight that MenSCs maintain a crosstalk with the physiologic environment, where their impact might modulate phenotypic and functional properties of innate immune populations —primarily macrophages—. Given the influence of the latter in the inflammatory context, MenSCs effects over macrophages must be taken into consideration when aiming to translate MenSCs to a clinical setting. On the other hand, the experimental models first described in this study are elegant work tools to analyse distinct aspects of MenSCs effects, and in general of MSCs, on specific severe pathologies., Tesis Univ. Granada.

In this doctoral thesis dissertation we describe the T lymphocyte mediated immune response, the inflammatory mechanisms, the endothelial cell adhesion molecules and the chemokines that mediate T lymphocyte cardiotropism during heart failure development, being many of these mechanisms potential targets for the treatment of this chronic disease., Tesis Univ. Granada. Programa Oficial de Doctorado en: Biomedicina

Tesis Univ. Granada. Departamento de Bioquímica y Biología Molecular III e Inmunología

In this doctoral thesis dissertation we describe the T lymphocyte mediated immune response, the inflammatory mechanisms, the endothelial cell adhesion molecules and the chemokines that mediate T lymphocyte cardiotropism during heart failure development, being many of these mechanisms potential targets for the treatment of this chronic disease., Tesis Univ. Granada. Programa Oficial de Doctorado en: Biomedicina

Tesis Univ. Granada. Departamento de Bioquímica y Biología Molecular III e Inmunología. Leída el 13 de octubre de 2011

Tesis Univ. Granada. Departamento de Bioquímica y Biología Molecular. Leída el 20 de marzo de 2006

Tesis Univ. Granada. Departamento de Bioquímica y Biología Molecular. Leída el 15 del 3 del 2007