Journal: bioRxiv

Article Title: Systemic Dosing of Virus-derived Serpin Improves Survival and Immunothrombotic Damage in Murine Colitis

doi: 10.1101/2024.01.08.574715

Figure Lengend Snippet: PEGSerp-1 treatment given as acute prophylaxis or as a chronic delated treatment significantly reduced iNOS positive M1 macrophage infiltrates in DSS colitis. Significantly reduced mean iNOS counts in 5% DSS with Acute prophylaxis PEGSerp-1 treatment ( A , ANOVA p < 0.0341, borderline p < 0.0587 Fishers PLSD for PEGSerp-1 vs Saline). iNOS stained 5% DSS histology sections illustrate loss of crypt architecture and extensive iNOS positive cell infiltrates with Saline treatments ( B ). There is reduced iNOS staining and improved crypt architecture with PEGSerp-1 treatment ( B ). Acute prophylaxis with PEGSerp-1 with 4% DSS did not significantly reduce iNOS staining ( C , p = 0.6765), however combined analysis of 4 and 5% PEGSerp-1 acute prophylaxis detected significantly reduced iNOS positive cell infiltrates (D, p < 0.0278). iNOS positive cell counts were significantly reduced with PEGSerp-1 treatment in both the 2%DSS Acute delayed (E, p < 0.0454) and the Chronic delayed treatment models ( F , p < 0.0005). Arg1+ M2 macrophage staining was not significantly modified by PEGSerp-1 in the 5% ( G , p = 0.2705), 4% ( H , p = 0.7007), nor the 2% acute ( I , p = 0.9552) and chronic delayed DSS colitis models ( J , p = 0.8766). IHC analysis performed at 20X, mean values calculated for 3 sections with visible positive staining. Micrographs – 20X. Red arrows indicate iNOS positive cells – IHC analysis.

Article Snippet: Immune cell markers assessed included iNOS+ M1 and Arginase 1+ M2 for macrophage; CD3 and CD4 for T lymphocytes, and Ly6G for neutrophils: nonspecific T lymphocytes (CD3, Abcam ab5690, 1:100 and CD4, Abcam, ab183685, 1:1,000), M1 macrophages (iNOS, Abcam ab15323, 1: 200), M2 macrophage arginase-1 (Cell Signaling, 93668, 1:200 and neutrophils (Ly6G, Invitrogen 1960181, 1:100).

Techniques: Saline, Staining, Modification

Journal: bioRxiv

Article Title: Systemic Dosing of Virus-derived Serpin Improves Survival and Immunothrombotic Damage in Murine Colitis

doi: 10.1101/2024.01.08.574715

Figure Lengend Snippet: PEGSerp-1 acute prophylaxis treatments in the 5% ( A, p = 0.6918) and 4% ( B, p = 0.7998) DSS colitis models, and the 2% chronic delayed ( D, p =) colitis models had no significant effect on Ly6G + neutrophil cell counts. In the acute delayes 2% DSS model, PEGSerp-1 tretament did significantly reduced Ly6G+ cell counts ( C, p < 0.05) Nonspecific CD3+ T cell counts were similarly unaffected by PEGSerp-1 treatments in the 5% ( E , p = 0.5757) and 4% ( F , p = 0.3894) acute prophylaxis and the 2% acute delayed colitis models ( G , p = 0.0869). CD4 + T cell counts were not modified by PEGSerp-1 treatment in the 5% ( H , p = 0.2556) and 4% ( I , p = 0.4481) acute prophylaxis models and in the 2% ( J, p = 0.0628) acute delayed model. However, CD4+ cell counts were significantly reduced with PEGSerp-1 treatment in the 2% chronic delayed colitis treatment model ( K , p < 0.0016).

Article Snippet: Immune cell markers assessed included iNOS+ M1 and Arginase 1+ M2 for macrophage; CD3 and CD4 for T lymphocytes, and Ly6G for neutrophils: nonspecific T lymphocytes (CD3, Abcam ab5690, 1:100 and CD4, Abcam, ab183685, 1:1,000), M1 macrophages (iNOS, Abcam ab15323, 1: 200), M2 macrophage arginase-1 (Cell Signaling, 93668, 1:200 and neutrophils (Ly6G, Invitrogen 1960181, 1:100).

Techniques: Modification

Journal: bioRxiv

Article Title: Systemic Dosing of Virus-derived Serpin Improves Survival and Immunothrombotic Damage in Murine Colitis

doi: 10.1101/2024.01.08.574715

Figure Lengend Snippet: Panel 1 - Micrographs illustrate reduced iNOS+ M1 macrophage positive staining in the submucosal vessels ( A -Saline, B -PEGSerp-1). iNOS positive counts are significantly reduced in the 5% DSS colitis model ( C , p < 0.0481), but not in the 2% chronic delayed treatment colitis model ( D , p = 0.5915). Arg1+ M2 macrophage positive cell counts ( E , p = 0.5915) and CD4+ T cell counts ( F, p = 0.1110) in the submucosal vessels were not altered in the 2% chronic delayed model. fXa+ cells ( G-I , p < 0.0331) and fibrinogen+ cells ( J, p < 0.0503) were significantly reduced in the 2% chronic delayed treatment model. uPAR positive staining was not reduced with PEGSerp-1 in the 5% acute prophylaxis (K, p = 0.1476) nor the 2% chronic delayed (L, p = 0.1675) models. C5b/9 positive cells in the 2% DSS Chronic delayed colitis model were borderline significantly reduced in submucosal vessels in the 2%chronic delayed colitis model with PEGSerp-1 treatment ( M , p = 0.0571). Dense perivascular aggregates of positively stained cells indicated by blue arrows. Mag 20X. Panel 2 – RT-PCR analysis of gene expression changes. uPAR expression significantly correlated with colon damage measured in the 2%DSS chronic delayed colitis model. Gene expression for fXa (B, p < 0.0035 ANOVA) and C3 (C, p < 0.0003ANOVA) were significantly increased in the cardiac sample extracts from the 2% DSS Chronic delayed model when compared to normal controls without DSS treatments. In the colon samples uPAR expression was increased, but not significantly (D, p = 08872). PAI-1 was significantly increased (E, p < 0.0172), and C1Inh showed a trend toward increased expression (F, p = 0.2238). In the heart no significant changes were detected for uPAR (G, p = 0.5587), PAI-1 (H, p = 0.3005)nor C1INH (I, p = 0.2708).

Article Snippet: Immune cell markers assessed included iNOS+ M1 and Arginase 1+ M2 for macrophage; CD3 and CD4 for T lymphocytes, and Ly6G for neutrophils: nonspecific T lymphocytes (CD3, Abcam ab5690, 1:100 and CD4, Abcam, ab183685, 1:1,000), M1 macrophages (iNOS, Abcam ab15323, 1: 200), M2 macrophage arginase-1 (Cell Signaling, 93668, 1:200 and neutrophils (Ly6G, Invitrogen 1960181, 1:100).

Techniques: Staining, Saline, Reverse Transcription Polymerase Chain Reaction, Expressing