Virus-like Particles with H5N1 Spikes and Paramyxovirus Cores for Diagnostics and Vaccination

open_access
Open Access
Author:
Ho, Caroline
Area of Honors:
Immunology and Infectious Disease
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Anthony Paul Schmitt, Thesis Supervisor
Girish Soorappa Kirimanjeswara, Thesis Honors Advisor
Keywords:
influenza
virus-like particles
nipah virus
Abstract:
Influenza viruses remain a major global health threat due to frequent antigenic variation and limited vaccine durability. Traditional influenza vaccines require annual reformulation and often provide suboptimal protection. Virus-like particles (VLPs) offer a promising alternative platform because they mimic native virions while remaining noninfectious. However, influenza VLPs based on the matrix 1 (M1) protein show low budding efficiency and inconsistent morphology. In contrast, matrix (M) proteins from paramyxoviruses such as Nipah virus (NiV) efficiently drive assembly and release of enveloped particles. This study investigated whether the NiV matrix (NiVM) protein can serve as a scaffold for producing hybrid influenza–paramyxovirus VLPs capable of incorporating influenza glycoproteins HEK-293T cells were transfected with plasmids encoding NiVM, influenza hemagglutinin (HA), and neuraminidase (NA). Cell lysates and supernatants were analyzed by SDS–PAGE and Western blot using anti-Myc, anti-FLAG, and anti-H5 primary antibodies with HRP-conjugated secondary detection. NiVM was consistently detected in all samples containing the M plasmid, confirming efficient particle formation. Both HA and NA were incorporated into VLP fractions when co-expressed with NiVM, whereas HA or NA expressed individually did not yield authentic VLP-associated signal. Although an NA band was observed in the NA-only condition, follow-up controls confirmed it resulted from nonspecific co-pelleted material rather than true particle formation. Cytoplasmic tail-swapping experiments using NiV and parainfluenza virus 5 (PIV5) domains did not enhance incorporation, indicating that the native influenza HA tail is compatible with NiVM-mediated assembly. These findings demonstrate that NiV M can independently mediate particle formation and recruit heterologous influenza glycoproteins, establishing proof of concept for cross-family hybrid VLP generation.