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Analysis of MLKL, RIP1 and RIP3 Immunostaining Markers in Human Liver Tissue from Fatal Yellow Fever Cases: Insights
Vanessa do Socorro Cabral Miranda1, Luiz Fabio Magno Falcão2, Hellen Thais Fuzii3
1Departmento of Pathology, Evandro Chagas Institute, Ministry of Health, Ananindeua 67030-000, PA, Brazil.
Abstract:
Necroptosis is a regulated form of cell death implicated in several pathological conditions, including viral infections. In this study, we investigated the expression and correlation of necroptosis markers MLKL, RIP1 and RIP3 in human liver tissue from fatal cases of yellow fever (YF) using immunohistochemistry (IHC). The liver samples were obtained from 21 YF-positive individuals and five flavivirus-negative controls with preserved liver parenchymal architecture. The cases underwent histopathological analysis, followed by tissue immunostaining with the immunohistochemical method of streptavidin-biotin peroxidase. Using the in situ method, we evaluated the centrilobular zone (Z3), midzonal zone (Z2), periportal zone and portal tract (PT) of human liver parenchyma with markers for necroptosis, RIPK1, RIPK3 and MLKL. A quantitative analysis revealed a significantly higher expression of MLKL, RIP1 and RIP3 in the liver parenchyma of YF cases compared to controls in different zones (Z3, Z2, Z1) and portal tracts (PTs) of the liver, especially in zone 2. Immunostaining confirmed the localization of MLKL, RIP1 and RIP3 in hepatocytes and inflammatory infiltrates, highlighting their involvement in the pathogenesis of YF. A Pearson correlation analysis demonstrated significant correlations among necroptosis markers, which indicates their coordinated regulation during YF-induced liver injury.
Insights
Necroptosis markers MLKL, RIP1, and RIP3 are significantly upregulated in the liver during fatal yellow fever (YF) infections. These necroptosis markers correlate with each other, indicating their role in YF-induced liver injury.
Area of Science:
- Hepatology
- Immunology
- Pathology
Background:
- Necroptosis, a programmed cell death pathway, is involved in viral infections and liver diseases.
- Yellow fever (YF) is a severe viral hemorrhagic fever with significant liver pathology.
Purpose of the Study:
- To investigate the expression and correlation of necroptosis markers (MLKL, RIP1, RIP3) in human liver tissue from fatal yellow fever cases.
- To determine the role of necroptosis in the pathogenesis of YF-induced liver injury.
Main Methods:
- Immunohistochemistry (IHC) was used to analyze liver tissue from 21 YF cases and 5 controls.
- Expression levels of MLKL, RIP1, and RIP3 were quantified in different liver zones (Z3, Z2, Z1) and portal tracts (PTs).
- Pearson correlation analysis was performed to assess the relationships among necroptosis markers.
Main Results:
- Significantly higher expression of MLKL, RIP1, and RIP3 was observed in YF liver tissue compared to controls, particularly in zone 2.
- Immunostaining confirmed the presence of these markers in hepatocytes and inflammatory infiltrates.
- Significant positive correlations were found among MLKL, RIP1, and RIP3, suggesting coordinated regulation.
Conclusions:
- Necroptosis is actively involved in the pathogenesis of yellow fever liver injury.
- The coordinated expression of MLKL, RIP1, and RIP3 highlights their collective role in YF-induced hepatocyte death.
- Targeting necroptosis pathways may offer therapeutic strategies for severe YF cases.
