Measurement of Lung Vessel and Epithelial Permeability In Vivo with Evans Blue

Smith, Prestina; Jeffers, Lauren A.; Koval, Michael

doi:10.1007/7651_2020_345

Cited by 17 publications

(13 citation statements)

References 15 publications

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“…5, M and N). This increased vascular permeability measured in CCR2 −/− MuHV-4–infected mice was further validated in MO-depleted mice by Evans blue quantification ( 47 ) in BALF and lung tissues after intravenous injection (Fig. 5O).…”

Section: Resultsmentioning

confidence: 59%

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Ly6C ^hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

et al. 2022

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Gammaherpesviruses (γHVs) have coevolved with their host, leading to a remarkably high infection prevalence and establishment of latency. The lifelong persistence of γHVs in hosts appears to broadly shape host immunity, and we show here that pulmonary infection with Murid herpesvirus 4 (MuHV-4), a mouse γHV, drives the recruitment of Ly6C hi monocytes (MOs) into the airway, thereby modulating the host immune response. The absence of Ly6C hi MOs is associated with severe virus-induced immunopathology and the systemic release of inflammatory mediators. Mechanistically, MuHV-4–imprinted MOs recruit CD4 T cells to the airways and trigger immunosuppressive signaling pathways through the PD-L1/PD-1 axis, thereby dampening the deleterious activation of cytotoxic CD4 T cells. These results uncover a role for Ly6C hi MOs in modulating CD4 T cell functions and reveal pathways that could be targeted therapeutically to reduce detrimental immunopathological responses associated with respiratory viral infections.

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Section: Resultsmentioning

confidence: 59%

“…BubbleGUM ( 47 ) analysis was used as described previously with default settings, and the gene sets (containing between 20 and 500 genes) from Gene Ontology and gene set were arbitrarily determined on the basis of the literature ( 10 ). The results with a false discovery rate (FDR) of ≤0.25 were considered as significant.…”

Section: Methodsmentioning

confidence: 99%

Ly6C ^hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

et al. 2022

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“…Specfically because the lungs are the most vulnerable and critical organ during septic shock, we measured lung vessel permeability by intravenous injection of Evans Blue dye to reflect the damage degree of the vascular endothelial barrier in the lung tissues of mice [ 29 , 30 ]. As shown in Figure 1 E, the accumulation of Evans Blue dye in lung tissues of mice induced by LPS (20 mg/kg) was obviously increased after LPS injection for 8 h ( Figure 1 E-3).…”

Section: Resultsmentioning

confidence: 99%

“…Lung vessel permeability was measured as previously described [ 30 , 57 ]. Briefly, mice were placed in a cage under the heat lamp for 5~10 min in order to dilate the blood vessels.…”

Section: Methodsmentioning

confidence: 99%

The Protective Effects of Goitrin on LPS-Induced Septic Shock in C57BL/6J Mice via Caspase-11 Non-Canonical Inflammasome Inhibition

et al. 2023

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Septic shock is defined as a subset of sepsis, which is associated with a considerably high mortality risk. The caspase-11 non-canonical inflammasome is sensed and activated by intracellular lipopolysaccharide (LPS) leading to pyroptosis, it plays a critical role in septic shock. However, there are few known drugs that can control caspase-11 non-canonical inflammasome activation. We report here that goitrin, an alkaloid from Radix Isatidis, shows protective effects in LPS-induced septic shock and significant inhibitory effect in caspase-11 non-canonical inflammasome pathway. Male C57BL/6J were injected intraperitoneally with LPS (20 mg/kg) to induce experimental septic shock. The results demonstrated that the survival rates of mice pretreated with goitrin or Toll-like receptor 4 (TLR4) inhibitor TKA-242 increased, and LPS-induced hypothermia and lung damage improved by inhibiting inflammatory response. Elucidating the detailed mechanism, we surprisingly found goitrin is really different from TAK-242, it independent of the TLR4 signal activation, but significantly inhibited the activation of caspase-11 non-canonical inflammasome, including cleaved caspase-11 and N-terminal fragment of gasdermin D (GSDMD-NT). Furthermore, with a nonlethal dose of the TLR3 agonist poly(I:C)-primed and subsequently challenged with LPS to induce caspase-11-mediated lethal septic shock, the efficacy of goitrin had been verified. Those results revealed the effect of goitrin in protective against LPS-induced septic shock via inhibiting caspase-11 non-canonical inflammasome, which provided a new therapeutic strategy for clinical treatment of septic shock.

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“…Evans blue dye was simultaneously used to measure the function of vascular and epithelial barriers of murine lungs in vivo. It has also been used as a marker of albumin clearance in murine models of acute lung injury [ 15 , 16 ]. In this study, we demonstrated that lung Evans blue staining could be used to assess the severity of air–blood barrier damage in Toxoplasma pneumonia.…”

Section: Discussionmentioning

confidence: 99%

Transcriptome analysis of a newly established mouse model of Toxoplasma gondii pneumonia

et al. 2023

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Background Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii. Toxoplasma gondii infection of the lungs can lead to severe pneumonia. However, few studies have reported Toxoplasma pneumonia. Most reports were clinical cases due to the lack of a good disease model. Therefore, the molecular mechanisms, development, and pathological damage of Toxoplasma pneumonia remain unclear. Methods A mouse model of Toxoplasma pneumonia was established by nasal infection with T. gondii. The model was evaluated using survival statistics, lung morphological observation, and lung pathology examination by hematoxylin and eosin (H&E) and Evans blue staining at 5 days post-infection (dpi). Total RNA was extracted from the lung tissues of C57BL/6 mice infected with T. gondii RH and TGME49 strains at 5 dpi. Total RNA was subjected to transcriptome analysis by RNA sequencing (RNA-seq) followed by quantitative real-time polymerase chain reaction (qRT–PCR) validation. Transcript enrichment analysis was performed using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases to assess the biological relevance of differentially expressed transcripts (DETs). Results C57BL/6 mice infected with T. gondii via nasal delivery exhibited weight loss, ruffled fur, and respiratory crackles at 5 dpi. The clinical manifestations and lethality of RH strains were more evident than those of TGME49. H&E staining of lung tissue sections from mice infected with T. gondii at 5 dpi showed severe lymphocytic infiltration, pulmonary edema, and typical symptoms of pneumonia. We identified 3167 DETs and 1880 DETs in mice infected with the T. gondii RH and TGME49 strains, respectively, compared with the phosphate-buffered saline (PBS) control group at 5 dpi. GO and KEGG enrichment analyses of DETs showed that they were associated with the immune system and microbial infections. The innate immune, inflammatory signaling, cytokine-mediated signaling, and chemokine signaling pathways displayed high gene enrichment. Conclusion In this study, we developed a new mouse model for Toxoplasma pneumonia. Transcriptome analysis helped to better understand the molecular mechanisms of the disease. These results provided DETs during acute T. gondii lung infection, which expanded our knowledge of host immune defenses and the pathogenesis of Toxoplasma pneumonia. Graphical Abstract

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Measurement of Lung Vessel and Epithelial Permeability In Vivo with Evans Blue

Cited by 17 publications

References 15 publications

Ly6C ^hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

Ly6C ^hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

The Protective Effects of Goitrin on LPS-Induced Septic Shock in C57BL/6J Mice via Caspase-11 Non-Canonical Inflammasome Inhibition

Transcriptome analysis of a newly established mouse model of Toxoplasma gondii pneumonia

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Measurement of Lung Vessel and Epithelial Permeability In Vivo with Evans Blue

Cited by 17 publications

References 15 publications

Ly6C hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

Ly6C hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

The Protective Effects of Goitrin on LPS-Induced Septic Shock in C57BL/6J Mice via Caspase-11 Non-Canonical Inflammasome Inhibition

Transcriptome analysis of a newly established mouse model of Toxoplasma gondii pneumonia

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Product

Resources

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Ly6C ^hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology

Ly6C ^hi monocytes balance regulatory and cytotoxic CD4 T cell responses to control virus-induced immunopathology