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Uptake of porcine rubulavirus (LPMV) by PK-15 cells
P Hernández-Jáuregui1, A Yacoub, S Kennedy
1Department of Veterinary Microbiology, Section of Virology, Swedish University of Agricultural Sciences, SLU, Biomedical Center, Uppsala, Sweden.
Background:
The porcine virus denominated La Piedad Michoacan Virus (LPMV) is a member of the family Paramyxoviridae and is the cause of a disease in pigs present only in Mexico. The disease is characterized by meningoencephalitis and respiratory distress in young pigs, epididymitis and orchitis in boars, and reproductive failure and abortion in sows.
Methods:
The cytopathology, morphology, and distribution of the hemagglutination neuraminidase (HN) and nucleoprotein (NP) proteins of LPMV were investigated following inoculation into PK-15 cells. The cytopathic effect was characterized by cytoplasmic vacuolation and the formation of syncytia and cytoplasmic inclusion bodies.
Results:
In immunofluorescence assays using a monoclonal antibody (MAb) against the HN protein at 5-60 min post-infection (early infection), a diffuse immunofluorescence was observed near the cell membrane and adjacent to the nuclear membrane. At 24 h post-infection (late infection), a dust-like immunofluorescence was observed throughout the cytoplasm. LPMV-infected cells incubated with the MAb against the NP protein showed punctate cytoplasmic fluorescence during the early stages of infection. At the late infection stage, these fluorescent particles became larger and were seen predominantly in the cytoplasm of syncytia. This pattern was also apparent by immunohistochemical labeling and immunogold electron microscopy. The latter technique revealed that HN protein was diffusely distributed throughout the cytoplasm. When using the MAb against the NP protein, nucleocapsid organization was the most prominent feature and resulted in the formation of cytoplasmic inclusion bodies visible by light and electron microscopy. Immunogold labeling of purified nucleocapsids was shown by electron microscopy. Virus particles and nucleocapsids were morphologically similar to members of the Paramyxoviridae family.
Conclusions:
The morphologic characteristics of the virions and the distribution patterns of the HN and NP proteins in PK-15 infected cells indicate that the mechanisms of LPMV replication are generally similar to those of the members of the Paramyxoviridae family.
Insights
La Piedad Michoacan Virus (LPMV), a Mexican swine paramyxovirus, causes significant disease. Studies reveal its replication mechanisms are similar to other paramyxoviruses, aiding in understanding swine paramyxovirus infections.
Area of Science:
- Veterinary Virology
- Molecular Biology
- Cell Biology
Background:
- La Piedad Michoacan Virus (LPMV) is a Mexican paramyxovirus causing disease in pigs.
- LPMV infection leads to neurological, respiratory, and reproductive issues in swine.
Purpose of the Study:
- To investigate the cytopathology, morphology, and protein distribution of LPMV in PK-15 cells.
- To understand LPMV replication mechanisms within the Paramyxoviridae family.
Main Methods:
- Inoculation of PK-15 cells with LPMV.
- Immunofluorescence assays using monoclonal antibodies against HN and NP proteins.
- Immunohistochemical labeling and immunogold electron microscopy.
Main Results:
- Cytopathic effects included cytoplasmic vacuolation, syncytia, and inclusion bodies.
- HN protein showed diffuse cytoplasmic distribution; NP protein formed punctate cytoplasmic fluorescence and nucleocapsid structures.
- Virus particles and nucleocapsids were morphologically consistent with Paramyxoviridae.
Conclusions:
- LPMV replication mechanisms are similar to other Paramyxoviridae family members.
- Protein distribution patterns provide insights into LPMV pathogenesis.
