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Generation of JC Polyoma Pseudovirus for High-Throughput Measurement of Neutralizing Antibodies
Mami Matsuda1, Tian-Cheng Li1, Akira Nakanishi2
1Department of Virology II, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.
Abstract:
Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system (CNS) caused by reactivation of dormant JC polyomavirus (JCPyV). PML was mainly observed in immunocompromised individuals, such as HIV-positive patients, autoimmune disease patients, and cancer patients. Given that the presence of anti-JCPyV antibodies in serum is a risk indicator for PML development, it is essential to monitor anti-JCPyV antibody levels. In the present study, we established reporter-based single-infection neutralization assays for JCPyV and the genetically similar BK polyoma virus (BKPyV). We then confirmed the lack of cross-reactivity between the two viruses using test sera obtained from mice immunized with plasmids encoding the JCPyV or BKPyV capsid. Next, we compared neutralization antibody titers in sera from healthy donors, patients with multiple sclerosis (MS), and HIV-positive patients using an in-house enzyme-linked immunosorbent assay (ELISA) with JCPyV-like particles (virus-like particles; VLPs). A positive correlation was demonstrated between the neutralization titer (75% infectious concentration; IC75) against JCPyV and the antibody titer obtained by VLP-based JCPyV ELISA. This assay system may be applied to detect antibodies against other PyVs by generation of pseudoviruses using the respective capsid expression plasmids, and is expected to contribute to the surveillance of PyV as well as basic research on these viruses.
Insights
Monitoring JC polyomavirus (JCPyV) antibodies is crucial for progressive multifocal leukoencephalopathy (PML) risk. New neutralization assays accurately detect JCPyV antibodies, aiding in PML surveillance and research.
Area of Science:
- Virology
- Immunology
- Neuroscience
Background:
- Progressive multifocal leukoencephalopathy (PML) is a CNS demyelinating disease caused by JC polyomavirus (JCPyV) reactivation.
- PML predominantly affects immunocompromised individuals, including those with HIV, autoimmune diseases, or cancer.
- Detecting anti-JCPyV antibodies is vital for assessing PML risk.
Purpose of the Study:
- To establish and validate reporter-based neutralization assays for JC polyomavirus (JCPyV) and BK polyomavirus (BKPyV).
- To compare JCPyV antibody levels in healthy donors, multiple sclerosis (MS) patients, and HIV-positive patients.
- To assess the correlation between neutralization and virus-like particle (VLP)-based ELISA antibody titers.
Main Methods:
- Developed reporter-based single-infection neutralization assays for JCPyV and BKPyV.
- Confirmed lack of cross-reactivity between JCPyV and BKPyV using mouse sera.
- Utilized an in-house ELISA with JCPyV VLPs to measure antibody titers in human sera.
Main Results:
- Established functional neutralization assays for JCPyV and BKPyV without cross-reactivity.
- Demonstrated a positive correlation between JCPyV neutralization titers (IC75) and VLP-ELISA antibody titers.
- Showcased the utility of the assay system for detecting antibodies in various patient cohorts.
Conclusions:
- The developed assay system is effective for detecting and quantifying JCPyV neutralizing antibodies.
- This system facilitates PML risk assessment and surveillance in at-risk populations.
- The platform can be adapted for PyV surveillance and fundamental research on polyomaviruses.
