Quantification of cell lysis by anidulafungin and caspofungin

ntly affect the apoE inhibition seen with IFN-a although the difference was not statistically significant. 16352702 T cell suppression of apoE production While apoE is believed to play an important protective role in atherosclerosis, the reverse is true for T cells. Through their secretion of cytokines such as IFN-c, activated T cells in atherosclerotic plaques may contribute to the inflammatory state that is characteristic of these lesions. To assess the potential interrelationship between monocytes and T cells, PBMC were simultaneously stimulated with TGF-b together with either antiCD3 or a pool of viral antigenic peptides to which most people have a T-cell response. As seen in figure 5, cultures in which T cells had been polyclonally activated showed strong IFNc responses and a more or less complete lack of apoE-secreting cells while the antigen-specific and lower IFN-c responses resulted in a more moderate inhibition of apoE secretion. ApoE production in macrophages ulated by LPS and IFN-c. However, compared to monocytes the effects on macrophages were modest and more clearly discerned with ELISA than ELISpot. As macrophages spontaneously produce not only apoE but also cytokines such as IL-6 and TNF-a, we conducted some preliminary experiments looking at to what extent these were produced by the same cells or not. For this purpose we used the FluoroSpot technique, a variant of the ELISpot which by Oleandrin employing fluorescent detection allows simultaneous detection of more than one secreted product. As shown in figure 7 with macrophages incubated 21821695 overnight in only medium, there were a similar number of cells secreting apoE and TNF-a. However, the two proteins were for the most part produced by separate cells with less than 20% of the apoE-producing cells also secreting TNF-a. In comparison, more than 70% of cells secreting the cytokine IL-6 also secreted TNF-a. As expected, addition of LPS to the cultures resulted in a reduced number of apoE-secreting cells whereas IL-6 and TNF-a were both strongly upregulated. However, concomitant with the reduced number of cells secreting apoE, the proportion of these cells also secreting TNF-a increased from 20 to about 50%. Four donors were tested and individual data are shown in ApoE production in hepatocytes As most apoE present in plasma has been reported to come from the liver, we investigated whether the same cytokines that Inflammation and apoE Production in Monocytes assays with a better capacity to detect minor differences in the amount of secreted apoE, differences that were not as easily discernible at the cellular level. To demonstrate the pros and cons of the two assays, different numbers of PBMC were stimulated with TGF-b and the number of secreting cells was determined in ELISpot while, in parallel cell cultures, the amount of apoE in cell supernatants was determined by ELISA. As shown in figure 8, apoE could be measured by ELISA with confidence at cell concentrations of 1006103 cells per well and above whereas in the ELISpot, secreting cells were readily detectable also at the lowest cell concentration of 12.56103 cells per well. This indicates a generally greater sensitivity of the ELISpot assay. However, the difference in apoE secretion between TGF-b-stimulated and unstimulated cells were more prominent in the ELISA. Together the two assays also demonstrate that the TGF-b-related increase in apoE production is likely the result of not only an increase in the number of secreting cells but also a