Journal: bioRxiv

Article Title: Toxoplasma induced cytokine release syndrome is critically dependent on the expression of pore-forming Perforin-Like Protein-1

doi: 10.1101/2025.03.17.643671

Figure Lengend Snippet: 10 2 WT (RH, red), 10 4 (green) RH Δplp1 , or 10 6 (blue) RH Δplp1 were administered i.p. to C57BL/6 mice, and total or percent neutrophils (CD45 + /CD11b int /Ly6G + ) present in the peritoneal cavity was measured by flow cytometry on days 3, 4, 5, 6, and 11 post-infection (A). Concentration of neutrophil growth factor G-CSF (B), and chemoattractants KC/CXCL1 (C) and MIP2/CXCL2 (D) were measured by Milliplex analysis in both serum and 2 mL peritoneal lavage wash throughout infection. Data are representative results from of two individual experiments with n=3 and 4 mice per group/PBS control n=1 mouse per day. * represents p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005, **** p ≤ 0.0001 **** represents p ≤ 0.001 as determined by two-way ANOVA. All WT infected mice succumbed to infection by days 8 and 9 post-infection.

Article Snippet: Additionally, a panel of 20 cytokines/chemokines were assessed using a custom MilliporeSigma Milliplex kit (EMD Millipore Corporation, Burlington MA); analytes included were IFN-gamma, IL-10, IL-12p70, IL-15, IL-1alpha, IL-2, IL-6, IP-10/CXCL10, KC/GRO/CINC-1/CXCL1, MCP-1/CCL2, MIP-1alpha/CCL3, MIP-1beta/CCL4, MIP-2/CXCL2, RANTES/CCL5, TNF-alpha, Eotaxin/CCL11, LIX, MIG/CXCL9, G-CSF, and GM-CSF.

Techniques: Flow Cytometry, Infection, Concentration Assay, Control

Journal: bioRxiv

Article Title: Toxoplasma induced cytokine release syndrome is critically dependent on the expression of pore-forming Perforin-Like Protein-1

doi: 10.1101/2025.03.17.643671

Figure Lengend Snippet: PEC lavage (A-E) and serum (F-J) cytokine and chemokine responses were measured and graphed for 10 4 Δplp1 (green), or 10 6 Δplp1 (blue) i.p. infection of C57BL/6 mice. IL-6 (A, F), IFN-g (B, G), MCP-1/CCL2 (C, H), MIP1-b/CCL4 (D, I), and G-CSF (E, J), were assessed by Milliplex between days 3 and 11 post-infection, as described. Graphs are representative results from one of two experiments (n=3 with a single PBS control per day). Data for RH infection is omitted to better demonstrate changes induced by PLP1 deficient parasite infection. Two-way ANOVA statistical analysis was used to determine significance; ** represents p ≤ 0.01, *** represents p ≤ 0.001.

Article Snippet: Additionally, a panel of 20 cytokines/chemokines were assessed using a custom MilliporeSigma Milliplex kit (EMD Millipore Corporation, Burlington MA); analytes included were IFN-gamma, IL-10, IL-12p70, IL-15, IL-1alpha, IL-2, IL-6, IP-10/CXCL10, KC/GRO/CINC-1/CXCL1, MCP-1/CCL2, MIP-1alpha/CCL3, MIP-1beta/CCL4, MIP-2/CXCL2, RANTES/CCL5, TNF-alpha, Eotaxin/CCL11, LIX, MIG/CXCL9, G-CSF, and GM-CSF.

Techniques: Infection, Control

Journal: bioRxiv

Article Title: Toxoplasma induced cytokine release syndrome is critically dependent on the expression of pore-forming Perforin-Like Protein-1

doi: 10.1101/2025.03.17.643671

Figure Lengend Snippet: Levels of IL-12p40 (A, E), IFN-γ (B, F), IL-18 (C, G), and TNF-α (D, H) in the PEC lavage (A-D) and serum (E-H) were assessed by cytokine ELISA and Milliplex on days 3, 4, 6, 8 and 11 post-infection. C57BL/6 mice were infected i.p. as previously described. Data are representative of two individual experiments with WT (RH 10 2 tachyzoites, red), low dose RH Δplp1 (10 4 tachyzoites, green), and high dose RH Δplp1 (10 6 tachyzoites, blue). PBS injected control mice (black) were included across the infection time course. Experiments were analyzed by two way ANOVA (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005, **** p ≤ 0.0001 ** represents p ≤ 0.01, *** represents p ≤ 0.001).

Article Snippet: Additionally, a panel of 20 cytokines/chemokines were assessed using a custom MilliporeSigma Milliplex kit (EMD Millipore Corporation, Burlington MA); analytes included were IFN-gamma, IL-10, IL-12p70, IL-15, IL-1alpha, IL-2, IL-6, IP-10/CXCL10, KC/GRO/CINC-1/CXCL1, MCP-1/CCL2, MIP-1alpha/CCL3, MIP-1beta/CCL4, MIP-2/CXCL2, RANTES/CCL5, TNF-alpha, Eotaxin/CCL11, LIX, MIG/CXCL9, G-CSF, and GM-CSF.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Injection, Control

Journal: bioRxiv

Article Title: Toxoplasma induced cytokine release syndrome is critically dependent on the expression of pore-forming Perforin-Like Protein-1

doi: 10.1101/2025.03.17.643671

Figure Lengend Snippet: PEC lavage (2 mL PBS wash of the peritoneal cavity, A-F) and serum (G-L) cytokine and chemokine responses were measured after i.p. infection of C57BL/6 mice with either 10 2 WT (RH, red), 10 4 RH Δplp1 (green), or 10 6 RH Δplp1 (blue) tachyzoites. IL-10 (A, G), IL-6 (B, H), MCP-1/CCL2 (C, I), MIP1-a/CCL3 (D, J), MIP1-b/CCL4 (E, K), and Eotaxin/CCL11 (F, L) were assessed by Milliplex between 3 and 11 dpi as described. Graphs are representative results from one of two experiments (n=3 with a single PBS control per day). Two-way ANOVA statistical analysis was used to determine significance; * represents p ≤ 0.05, ** represents p ≤ 0.01, *** represents p ≤ 0.001. All WT infected mice succumbed by days 8 and 9 post-infection.

Article Snippet: Additionally, a panel of 20 cytokines/chemokines were assessed using a custom MilliporeSigma Milliplex kit (EMD Millipore Corporation, Burlington MA); analytes included were IFN-gamma, IL-10, IL-12p70, IL-15, IL-1alpha, IL-2, IL-6, IP-10/CXCL10, KC/GRO/CINC-1/CXCL1, MCP-1/CCL2, MIP-1alpha/CCL3, MIP-1beta/CCL4, MIP-2/CXCL2, RANTES/CCL5, TNF-alpha, Eotaxin/CCL11, LIX, MIG/CXCL9, G-CSF, and GM-CSF.

Techniques: Infection, Control

Journal: Scientific Reports

Article Title: Proinflammatory cytokines, oxidative stress, and organ function as biomarkers of soman (GD) chronic neurotoxicity

doi: 10.1038/s41598-025-94190-z

Figure Lengend Snippet: Secreted pro-inflammatory cytokine/chemokines in the CSF and serum of the rats at 18 weeks post-soman exposure. A customized MILLIPLEX® Rat Cytokine/Chemokine kit was used to analyze each cytokine/chemokine. The results are plotted as pg/mL of CSF ( a ) or serum ( b ) for TNF-α, IL-6, IL-17A, MCP-1, IL-18, and IL-1α. Normality was assessed with the Shapiro–Wilk test. Bars represent mean ± SEM, n = 10 (5/sex) for serum and n = 8 (4/sex) for CSF. ** p < 0.01, *** p < 0.001 versus control by Unpaired t-test (MCP-1) and * p < 0.05, ** p < 0.01 and *** p < 0.001, **** p < 0.0001 versus control by Mann–Whitney test (TNF-α, IL-6, IL-17A, IL-18, and IL-1α). No sex differences were observed, and data are presented as mixed-sex cohort.

Article Snippet: Customized MILLIPLEX® Rat Cytokine/Chemokine kit (Millipore Sigma, RECYTMAG-65K-08C) was used to assess the levels of TNF-α, IL-6, IL-1α, IL-18, IL-17A, and MCP-1.

Techniques: Control, MANN-WHITNEY