Journal: Cell Death & Disease
Article Title: Neutrophil extracellular traps characterize caseating granulomas
doi: 10.1038/s41419-024-06892-3
Figure Lengend Snippet: We detected NETs by the quantification of circulating NET degradation products and found that they are elevated in patients with TB when compared to NHD. a We quantified cfDNA in the sera by PicoGreen fluorescence, b MPO–DNA and c NE-DNA complexes by ELISA and d NE activity by the conversion of a specific fluorogenic substrate. e Summarizes mean values and ranges for all parameters. f – m Clinical parameters were associated with the results obtained by NET formation analyses of TB sera. f NE-DNA complexes, g MPO–DNA complexes, h NE activity and i neutrophil counts in the sera of TB patients, associated with the type of TB case: new (green) versus relapse (red). Note that NET formation is increased in the sera of TB patients in relapse. j NE-DNA complexes, k MPO–DNA complexes, l NE activity and m neutrophil counts in the sera of TB patients, grouped by the extent of tissue destruction: no destruction (green) or destruction (red). Note that NET formation, NE activity and neutrophil counts are increased in patients with tissue destruction. All statistical analyses were performed employing the Mann–Whitney test. Data are presented as mean ± standard deviation (s.d.). At least three technical replicates were obtained for each data set. NHD normal healthy donors, TB tuberculosis, MPO myeloperoxidase, NE neutrophil elastase, MFI mean fluorescence intensity, n.s. not significant.
Article Snippet: To assess the activity of NE in the sera of patients with TB, 10 µL of serum and 100 µM fluorogenic NE substrate (MeOSuc-AAPV-AMC, sc-201163, Santa Cruz Biotechnology Inc., Dallas, TX, USA) per well were diluted in Dulbecco´s phosphate buffered saline (DPBS, Thermo Fisher Scientific Inc., Waltham, MA, USA) in a 384-well plate (#3680, Corning Inc., Corning, NY, USA).
Techniques: Fluorescence, Enzyme-linked Immunosorbent Assay, Activity Assay, MANN-WHITNEY, Standard Deviation
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