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. 2022 Nov 17;55(12):2436–2453.e5. doi: 10.1016/j.immuni.2022.11.007

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© 2022 The Authors

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Splenic cell death and reduced plasma DNA degradation capacity in sepsis and COVID-19 pneumonia

(A) Representative immunofluorescence confocal micrographs of post-mortem spleens from 1 control and 9 individuals with CP, stained for T cells (CD3, cyan), TUNEL (magenta), and myeloperoxidase (MPO, neutrophils; yellow). Scale bars: 100 μm.

(B) High-magnification micrographs from (A). Scale bars: 20 μm.

(C) Quantification of the percentage of TUNEL⁺ cells that are CD3⁺ in the splenic tissues shown in (A) (left) and distribution of TUNEL⁺ T cells (CD3⁺), neutrophils (MPO⁺), or other cells in (A) (right).

(D) Correlation between plasma actin (ACTA2; ACTB; ACTG1) relative protein abundance (RPA) and blood lymphocyte counts in the final sample obtained from 63 participants with CP who reached WHO-7.

(E) Plasma DNA in healthy (HD, gray), CP (green), and sepsis participants (SP, magenta) or mice infected with C. albicans (SM, yellow).

(F) Plasma samples from HD, SP, CP, and SM immunoblotted for actin and histone H2B.

(G) Correlation between actin abundance measured by densitometric analysis of the western immunoblots in (F) or mass spectrometry in SP plasma.

(H) Correlation between SP plasma actin and DNA.

(I) Correlation between histone H3 measured by western immunoblot (left) or actin (right) and platelet factor 4 (PF4) measured by mass spectrometry.

(J) Plasmid DNA degradation by 30 HD, 40 CP, and 36 SP plasmas. D₅₀ values were calculated from raw data in Figures S2D and S2E. Low activity range (D₅₀ > 15) depicted by the purple bar on the y axis.

(K) NETase activity in 3% plasma from 30 HD, 87 CP, and 36 SP measured by timelapse microscopy as the loss in human NET DNA intensity over 8 h.

(L) Correlation between DNase activity at 10% plasma dilution and NETase activity in CP plasmas.

(M) DNase I concentrations in HD (blue) or CP plasmas with WHO-3, -4 (green), or -7 (orange) and SP plasmas (pink). Statistics by Mann-Whitney or Kruskal-Wallis test for single comparisons, one-way ANOVA for multiple comparisons, simple linear or non-linear regression for correlations. See also Figure S2.