A causal relationship between plasma ceramide concentration and respiratory distress symptoms in COVID-19 patients is inferred. In this study, plasma samples of 52 individuals infected with COVID-19 were utilized in a lipidomic analysis. Lipids belonging to the ceramide class exhibited a 400-fold increase in total plasma concentration in infected patients. Further analysis led to the demonstration of concentration dependency for severe COVID-19 respiratory symptoms in a subclass of ceramides. The subclasses Cer(d18:0/24:1), Cer(d18:1/24:1), and Cer(d18:1/22:0) were shown to be increased by 48-, 40-, and 33-fold, respectively, in infected plasma samples and to 116-, 91- and 50-fold, respectively, in plasma samples with respiratory distress. Hence, monitoring plasma ceramide concentration, can be a valuable tool for measuring effects of therapies on COVID-19 respiratory distress patients.

Cultured porcine aortic endothelium was transfected with sequences that encode the SV40 T antigen, resulting in an immortalized cell line that retains the differentiated properties of normal aortic endothelial cells. Specifically, these cells form cobblestone monolayers, synthesize von Willebrand factor, and endocytose acetylated LDL. These cells can be readily propagated in culture and have been passaged over a year in culture. The cells form tubular structures when grown on Matrigel. The cells in culture express class I but do not express MHC class II antigens. Both class I and class II expression can be induced by treatment with recombinant swine interferon‐γ. Expression of CD44 and several other cell surface antigens have been observed. Co‐culture of these cells with purified human CD4+ T cells resulted in a significant T‐cell proliferative response similar to that observed for primary porcine EC. Finally, the cells are readily susceptible to transfection and express exogenous genes. These cells should be valuable for study of human anti‐porcine endothelial responses.

The complexity of IFN-mediated regulation of the murine Ly-6E gene in T cell lines is highlighted by the following observations: 1) multiple regulatory regions are present within different parts of the Ly-6E promoter and are necessary for IFN inducibility of the Ly-6E gene, 2) multiple transcription factors including Oct-1 and Oct-2 and the high mobility group (HMG) protein HMGI(Y) bind to regulatory elements present within the G region required for both IFN-αβ and IFN-γ responses, 3) mutational analysis of the G region reveals that a complex interaction exists between the factors binding to this region as shown by their mutual interdependence for detection in DMSA, and 4) inhibition of expression of HMG proteins by antisense HMGI-C RNA in EL4 cells causes the loss of IFN-αβ and IFN-γ inducibility of the endogenous Ly-6 gene. These findings taken together suggest that, in response to IFN treatment, an HMG protein-dependent complex involving multiple regulatory factors is assembled and is required for IFN inducibility of the Ly-6E gene.

The murine Ly6-E gene is transcriptionally induced by interferon-α/β (IFN-α/β) and IFN-γ in a variety of distinct cell types. The mechanism of IFN inducibility in B-cell lines was investigated by deletion analysis of the promoter and by identifying DNA binding proteins in mobility shift assays. A region located in the distal part of the promoter at −2.3 kb contributed to inducibility by both types of IFNs. This region contains a novel element in addition to the previously well-characterized IFN-stimulated response element (ISRE). The probes containing ISRE detected IFN-inducible complexes in mobility shift assays and the signal transducer and activator of transcripition–1 was found to be in these complexes from cells treated with either type of IFN. An additional element present in the proximal part of the promoter at position −109 is also required for IFN-α/β–mediated induction. These data suggested a cooperative interaction between these physically disparate regulatory regions. A crucial role for HMGI(Y) protein in this cooperative multiprotein complex is supported by the evidence that inhibition of HMGI(Y) expression via antisense RNA results in the loss of IFN-α/β–mediated induction of the Ly6-E gene. These results show the complexity involved in achieving cell-type specificity in IFN-mediated gene regulation.

The murine Ly6-E gene is transcriptionally induced by interferon-α/β (IFN-α/β) and IFN-γ in a variety of distinct cell types. The mechanism of IFN inducibility in B-cell lines was investigated by deletion analysis of the promoter and by identifying DNA binding proteins in mobility shift assays. A region located in the distal part of the promoter at −2.3 kb contributed to inducibility by both types of IFNs. This region contains a novel element in addition to the previously well-characterized IFN-stimulated response element (ISRE). The probes containing ISRE detected IFN-inducible complexes in mobility shift assays and the signal transducer and activator of transcripition–1 was found to be in these complexes from cells treated with either type of IFN. An additional element present in the proximal part of the promoter at position −109 is also required for IFN-α/β–mediated induction. These data suggested a cooperative interaction between these physically disparate regulatory regions. A crucial role for HMGI(Y) protein in this cooperative multiprotein complex is supported by the evidence that inhibition of HMGI(Y) expression via antisense RNA results in the loss of IFN-α/β–mediated induction of the Ly6-E gene. These results show the complexity involved in achieving cell-type specificity in IFN-mediated gene regulation.

A causal relationship between plasma ceramide concentration and respiratory distress symptoms in COVID-19 patients is presented. In this study, plasma samples of 52 individuals infected with COVID-19 were utilized in a lipidomic analysis. Lipids belonging to the ceramide class exhibited a 400-fold increase in total plasma concentration in infected patients. Further analysis led to the demonstration of concentration dependency for severe COVID-19 respiratory symptoms in a subclass of ceramides. The subclasses Cer(d18:0/24:1), Cer(d18:1/24:1), and Cer(d18:1/22:0) were shown to be increased by 48-, 40-, and 33-fold, respectively, in infected plasma samples and to 116-, 91- and 50-fold, respectively, in plasma samples with respiratory distress. Hence, monitoring plasma ceramide concentration, targeting ceramide synthesis, its salvage and its regulatory mechanisms are validated approaches towards enhancing survival from COVID-19 respiratory distress.

A causal relationship between plasma ceramide concentration and Covid-19 patients with respiratory distress symptoms is presented. Hence, monitoring of plasma ceramide concentration, targeting ceramide synthesis, its salvage and its regulatory mechanisms, are validated approach towards enhancing survival from Covid-19 respiratory distress. In this study, plasma samples of 52 individuals infected with Covid-19 were utilized in a lipidomic analysis. Lipids belonging to ceramide class exhibited a 400-fold increase in total plasma concentration in infected patients. Further analysis lead to demonstration of concentration dependency, for severe Covid-19 respiratory symptoms, in a subclass of ceramides. The subclasses Cer(d18:0/24:1), Cer(d18:1/24:1), and Cer(d18:1/22:0) are shown to be increased by 48, 40, and 33–folds respectively in infected plasma samples, and to 116, 91 and 50-folds in plasma samples with respiratory distress.

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