Journal: Frontiers in Immunology

Article Title: Inflammatory response and parasite regulation in acute toxoplasmosis: the role of P2X7 receptor in controlling virulent atypical genotype strain of Toxoplasma gondii

doi: 10.3389/fimmu.2024.1452828

Figure Lengend Snippet: The P2X7 receptor controls the parasite load in C57BL/6 mice. (A) Fluorescence images were acquired using four WT-infected mice and five P2X7 -/– infected mice. (B) averaged radiance in equal-sized regions of interest (ROI). Mice were placed inside the IVIS lumina chamber and imaged in the ventral, and the fluorescence was measured in the ROI. (C) The B1 gene expression was analyzed by qPCR. (D) Representative photomicrograph of a plaque assay with LLC-MK2 cells. (E) Quantification of proliferation parasite area. (B) Data represent the mean and SD of four WT mice and five P2X7 -/- mice analyzed by unpaired t-test. (C) Data represent the mean and SD of four WT mice and three P2X7 -/- mice analyzed by unpaired t-test. (D) Data represent the mean and SEM of forty-three photos from three animals analyzed by unpaired t-test. (*) Significance compared to the WT group. (*) p < 0.05; (**) p < 0.01.

Article Snippet: The real-time PCR (qRT-PCR) was performed using HOT FIREPol ® EvaGreen ® qPCR Mix Plus (Promega) to analyze the following targets: NLRP3, IL-1β, TNF, IFN-γ, Foxp3, and β-actin ( ).

Techniques: Fluorescence, Infection, Gene Expression, Plaque Assay

Journal: Frontiers in Immunology

Article Title: Inflammatory response and parasite regulation in acute toxoplasmosis: the role of P2X7 receptor in controlling virulent atypical genotype strain of Toxoplasma gondii

doi: 10.3389/fimmu.2024.1452828

Figure Lengend Snippet: The EGS strain infection changes the inflammatory parameters and subpopulation cells in the intestine of infected animals. (A–D, F) RT-qPCR analyzed the NLRP3, IL-1β, TNF-α, IFN-γ, and Foxp3 gene expression. (E) Secretion levels of IFN-γ were analyzed in the supernatant in the ileum explant. (G) The gating strategy of T reg population (CD4 + Foxp3 + ) cells analysis in lamina propria. (H) The histogram showed the CD4 + Foxp3 + cell. (I) Quantification of CD4 + Foxp3 + cells. (A–F) Data represent the mean and SD of 3 - 6 animals. Analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons test. (I) Data represent the mean and SD of 4 WT, WT-infected, and P2X7 -/- mice and 7 P2X7 -/– infected mice. Analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons test. (*) Significance compared to the non-infected group; (#) significance between infected groups. (*/#) p < 0.05; (**) p < 0.01; (***) p < 0.001; (###) p < 0.001; (****/####) p < 0.0001.

Article Snippet: The real-time PCR (qRT-PCR) was performed using HOT FIREPol ® EvaGreen ® qPCR Mix Plus (Promega) to analyze the following targets: NLRP3, IL-1β, TNF, IFN-γ, Foxp3, and β-actin ( ).

Techniques: Infection, Quantitative RT-PCR, Gene Expression

Journal: Frontiers in Immunology

Article Title: Inflammatory response and parasite regulation in acute toxoplasmosis: the role of P2X7 receptor in controlling virulent atypical genotype strain of Toxoplasma gondii

doi: 10.3389/fimmu.2024.1452828

Figure Lengend Snippet: The EGS strain promotes liver damage and liver inflammatory infiltration. (A) Liver photomicrography, the asterisks show points of necrosis in the tissue. (B) liver weight weighed. (C) The B1 gene expression by qPCR. (D) Blood levels of ALT for systemic and liver tissue damage. (E) Photomicrography of liver tissue stained with hematoxylin and eosin. (F) Graphic showing liver inflammation index quantification. (G) Graphic showing granuloma area mensure. (B) Data represent the mean and SD of 13 WT and 10 P2X7 -/- uninfected mice, and 19 WT and 20 P2X7 -/– infected mice in three different experiments, analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons test. (C) Data represent mean and SD of three or four animals analyzed by unpaired t-test. (D) Data represent the mean and SD of 4 uninfected mice, 11 WT-infected mice, and 8 P2X7 -/– infected mice in two different experiments, analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons test. (F-H) Data represent the mean and SEM of seventeen images from three animals analyzed by unpaired t-test. (*) Significance compared to the non-infected group; (#) significance compared between infected groups. (#) p < 0.05; (**) p < 0.01; (***) p < 0.001; (****/) p < 0.0001.

Article Snippet: The real-time PCR (qRT-PCR) was performed using HOT FIREPol ® EvaGreen ® qPCR Mix Plus (Promega) to analyze the following targets: NLRP3, IL-1β, TNF, IFN-γ, Foxp3, and β-actin ( ).

Techniques: Gene Expression, Staining, Infection

Journal: Frontiers in Immunology

Article Title: Inflammatory response and parasite regulation in acute toxoplasmosis: the role of P2X7 receptor in controlling virulent atypical genotype strain of Toxoplasma gondii

doi: 10.3389/fimmu.2024.1452828

Figure Lengend Snippet: The P2X7 receptor controls the T. gondii parasite proliferation in mice spleen. (A) Spleen photomicrography. (B) Graphic of spleen weight. (C) Representative photomicrograph of a plaque assay with LLC-MK2 cells. Scale bar 50 µm. (D) Quantification of proliferation parasite area. (E) Parasite load quantification using the B1 gene expression analyzed by qPCR. (F, G) Levels of IL-1β, and IFN-γ cytokines in the supernatant in splenocytes culture after 72 (h). (B) Data represent the mean and SD of 13 WT and 10 P2X7 -/- uninfected mice, and 19 WT and 20 P2X7 -/– infected mice in three different experiments, analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons test. (D) Data represent the mean, and SEM of forty-three photos from three different animals analyzed by unpaired t-test. (E) Data represent the mean and SD of 3 - 6 animals, analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons tests. (F, G) Data represent the mean and SD of 3 - 6 animals, analyzed by ordinary one-way ANOVA, Tukey’s multiple comparisons test. (*) Significance compared to the non-infected group. (**) p < 0.01; (****) p < 0.0001.

Article Snippet: The real-time PCR (qRT-PCR) was performed using HOT FIREPol ® EvaGreen ® qPCR Mix Plus (Promega) to analyze the following targets: NLRP3, IL-1β, TNF, IFN-γ, Foxp3, and β-actin ( ).

Techniques: Plaque Assay, Gene Expression, Infection