Project description:Eighteen patients admitted to CSMC and diagnosed with COVID19 by RT-PCR were stratified into COVID19 mild/moderate, severe, and recovery groups (n=5-6/group). Venous blood was collected into EDTA coated tubes and centrifuged to separate plasma and buffy coat. Plasma was collected and frozen at -80C, and the buffy coat was collected into cryo-preservation media and frozen at -80C. Recovered cells are sorted for live-dead staining, then fixed with methanol. Fixed single cells were further captured using 10x chromium Next GEM 3 prime v3.1 kit. Two patients samples from the same group were mixed, captured and sequenced together.
Project description:Single cell RNAseq was performed on PBMC isolated from health workers who received two doses of Pfizer COVID19 vaccine and determined as either normal responders or low responders by antibody test. Each group has 4 patients. Cells from each patient were labeled with Totalseq C hashing antibodies and mixed 1:1:1:1 for each group. Single cells were further captured using 10x chromium Next GEM 5 prime kit.
Project description:Objective: COVID19 is caused by the SARS-CoV-2 virus and has been associated with severe inflammation leading to organ dysfunction and mortality. Our aim was to profile the transcriptome in leukocytes from critically ill ICU patients positive for COVID19 vs. those negative for COVID19 to better understand the COVID19 associated host response. Design: Transcriptome profiling of buffy coat cells via ribonucleic acid sequencing (RNAseq) at the time of admission to the ICU. Setting: Tertiary care ICU and academic laboratory. Subjects: All patients admitted to the ICU suspected of being infected with SARS-CoV-2, using standardized hospital screening methodologies, had blood samples collected at the time of admission to the ICU. Interventions: None. Measurement and Main Results: Age- and sex-matched ICU patients that were either COVID19+ (PCR positive, 2 genes) or COVID19- (PCR negative) were enrolled. Cohorts were well-balanced with the exception that COVID19- patients had significantly higher total white blood cell counts and circulating neutrophils and COVID19+ patients were more likely to suffer bilateral pneumonia compared to COVID19- patients. Further, the mortality rate for this cohort of COVID19+ ICU patients was 29%. Transcriptional analysis revealed that when compared to COVID19- patients, the altered transcriptional responses of leukocytes in critically ill COVID19+ ICU patients appeared to be associated with multiple interrelated outcomes, including but not limited to robust interferon (IFN)-associated transcriptional responses, a marked decrease in the transcriptional activity of genes contributing to protein synthesis and the dysregulated expression of genes that contribute to coagulation, platelet activation, Toll-like receptor activation, neurotrophin signaling, and protein SUMOylation/ubiquitination. Conclusions: COVID19+ patients on day 1 of admission to the ICU display a unique leukocyte transcriptional profile that distinguishes them from COVID19- patients. Identification of this profile provides guidance for future targeted studies exploring novel prognostic/therapeutic aspects of COVID19.
Project description:High-resolution mass spectrometry and the parallel quantitative evaluation of thousands of proteins has been used to characterise the proteomes of peripheral blood neutrophils from >200 individuals. This work has comprehensively mapped neutrophil molecular changes associated with mild versus severe COVID19 and identified significant quantitative changes in more than 1700 proteins in neutrophils from patients hospitalised with COVID19 versus patients with non-COVID19 acute respiratory infections. The study identifies neutrophil protein signatures associated with COVID19 disease severity. The data also show that alterations in neutrophil proteomes can persist in fully recovered patients and identify distinct neutrophil proteomes in recovered versus non recovered patients. Our study provides novel insights into neutrophil responses during acute COVID-19 and reveal that altered neutrophil phenotypes persist in convalescent COVID19 infections.
Project description:Mass Spectrometry identifies temporal changes and hallmarks of Delayed Recovery in the COVID19 Neutrophil proteomes MTD project_description High-resolution mass spectrometry and the parallel quantitative evaluation of thousands of proteins has been used to characterise the proteomes of peripheral blood neutrophils from >200 individuals. This work has comprehensively mapped neutrophil molecular changes associated with mild versus severe COVID19 and identified significant quantitative changes in more than 1700 proteins in neutrophils from patients hospitalised with COVID19 versus patients with non-COVID19 acute respiratory infections. The study identifies neutrophil protein signatures associated with COVID19 disease severity. The data also show that alterations in neutrophil proteomes can persist in fully recovered patients and identify distinct neutrophil proteomes in recovered versus non recovered patients. Our study provides novel insights into neutrophil responses during acute COVID-19 and reveal that altered neutrophil phenotypes persist in convalescent COVID19 infections.
Project description:Background and Objectives Recently accumulating evidence demonstrates the rare occurrence of COVID19 vaccination- induced inflammation in the central nervous system. However, the precise immune dysregulation related to the COVID19 vaccination-associated autoimmunity remains elusive. Here we report a case of COVID19 vaccination-associated encephalitis, where single-cell RNA sequencing (scRNA-seq) analysis identified a distinct myeloid cell population in the blood during the acute phase of the disease. Methods Peripheral blood mononuclear cells (PBMCs) were analyzed by scRNA-seq to clarify the cellular components of the patients at both the acute and remission phases of the disease. The data obtained were compared to those acquired from a healthy cohort. Results In this study, scRNA-seq analysis of PBMCs identified a distinct myeloid cell population observed at the acute phase of encephalitis. The specific myeloid population was not detected both at the remission phase of the disease and the samples fromof a healthy cohort. Discussion Our findings illustrate the induction of a unique myeloid subset in COVID19 vaccination-associated encephalitis. Further accumulation of reports investigating the dysregulated immune signature of COVID19 vaccination-associated autoimmunity is warranted to clarify the pathogenic role of the myeloid subset observed in our study.
Project description:Background and Objectives Recently accumulating evidence demonstrates the rare occurrence of COVID19 vaccination- induced inflammation in the central nervous system. However, the precise immune dysregulation related to the COVID19 vaccination-associated autoimmunity remains elusive. Here we report a case of COVID19 vaccination-associated encephalitis, where single-cell RNA sequencing (scRNA-seq) analysis identified a distinct myeloid cell population in the blood during the acute phase of the disease. Methods Peripheral blood mononuclear cells (PBMCs) were analyzed by scRNA-seq to clarify the cellular components of the patients at both the acute and remission phases of the disease. The data obtained were compared to those acquired from a healthy cohort. Results In this study, scRNA-seq analysis of PBMCs identified a distinct myeloid cell population observed at the acute phase of encephalitis. The specific myeloid population was not detected both at the remission phase of the disease and the samples fromof a healthy cohort. Discussion Our findings illustrate the induction of a unique myeloid subset in COVID19 vaccination-associated encephalitis. Further accumulation of reports investigating the dysregulated immune signature of COVID19 vaccination-associated autoimmunity is warranted to clarify the pathogenic role of the myeloid subset observed in our study.