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Persistent measles virus infection of the intestine: confirmation by immunogold electron microscopy
1Inflammatory Bowel Disease Study Group, Royal Free Hospital School of Medicine, London.
Abstract:
This study sought to investigate persistent measles virus infection of the intestine: a novel protocol for immunogold electron microscopy was developed using a polyclonal anti-measles nucleoprotein antibody on reprocessed, formalin fixed paraffin wax embedded tissue sections. Antibody binding was detected using both immunoperoxidase and light microscopy on tissue sections, and 10 nm gold conjugated secondary antibody and electron microscopy on ultrathin sections. The techniques were validated using both measles infected vero cells and human tissues with established measles infection: these included brain affected by subacute sclerosing panencephalitis and acute measles appendicitis. The technique was applied subsequently to six untreated cases of granulomatous Crohn's disease, and two cases of ileocaecal tuberculosis, a granulomatous control. Mumps primary antibody--applied to both mumps infected vero cells, and measles infected vero cells and tissues studied by immunoperoxidase, and measles antibody on mumps infected cells studied by immunoperoxidase and immunogold--were used as specificity controls: the primary antibodies identified their respective target antigen and there was no antibody cross reactivity. Measles virus nucleocapsids labelled with gold conjugated antibody in both infected cells and tissues, including foci of granulomatous inflammation in five of six cases of Crohn's disease: in the fifth case, the granuloma could not be identified in ultrathin section. In one of the tuberculosis cases, a low level of signal was noted while the second case was negative. Labelling adopted a characteristic pattern in all infected tissues, strengthening the specificity of these findings. This study provides the first direct confirmation of persistent measles virus infection of the intestine.
Insights
Persistent measles virus infection in the intestine was confirmed using a novel immunogold electron microscopy protocol. This groundbreaking technique provides direct evidence of measles virus nucleocapsids in intestinal tissues, including Crohn
Area of Science:
- Gastroenterology
- Virology
- Immunohistochemistry
Background:
- The role of persistent viral infections in chronic gastrointestinal diseases remains an area of active investigation.
- Measles virus is known for its ability to establish persistent infections in various host tissues.
- Direct visualization of viral components within intestinal tissue has been challenging.
Purpose of the Study:
- To develop and validate a novel immunogold electron microscopy protocol for detecting persistent measles virus infection in the intestine.
- To investigate the presence of measles virus in granulomatous inflammation associated with Crohn's disease and tuberculosis.
- To provide direct evidence of measles virus nucleocapsids in intestinal tissues.
Main Methods:
- Development of a novel immunogold electron microscopy protocol using a polyclonal anti-measles nucleoprotein antibody.
- Reprocessing of formalin-fixed paraffin-embedded (FFPE) tissue sections for enhanced antibody binding.
- Validation of the technique using measles-infected cells and human tissues (subacute sclerosing panencephalitis, acute measles appendicitis), with specificity controls using mumps virus.
Main Results:
- The novel protocol successfully detected measles virus nucleocapsids in infected cells and human tissues.
- Measles virus nucleocapsids were identified in foci of granulomatous inflammation in five out of six Crohn's disease cases.
- A low signal was observed in one tuberculosis case, while the other was negative, indicating measles virus is not a common finding in ileocecal tuberculosis.
Conclusions:
- This study presents the first direct confirmation of persistent measles virus infection in the human intestine.
- The developed immunogold electron microscopy technique is effective for visualizing viral nucleocapsids in FFPE intestinal tissues.
- The findings suggest a potential role for measles virus in the pathogenesis of granulomatous inflammation in Crohn's disease.
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