Virus-Like Particles Assembled Using Respiratory Syncytial Virus Matrix Protein Elicit Protective Immunity in Mice

Abstract

Heterologous virus-like particle (VLP) assembly involving influenza or the Newcastle disease virus matrix protein (M) has been extensively used to explore the efficacies of VLP vaccines against the respiratory syncytial virus (RSV). Here, we attempted to generate homologous RSV VLPs by expressing the pre-fusion (pre-F) or the glycoprotein (G) on the RSV M protein and evaluated their protective efficacy in mice. Methods: We generated VLPs using the baculovirus expression system in Spodoptera frugiperda (Sf9) insect cells. Recombinant baculoviruses expressing the RSV pre-F, G, and M antigens were inoculated into Sf9 cells, and particles were self-assembled. Mice were immunized with either pre-F or G-expressing VLPs, and immune parameters were assessed to determine protection. Results: Our findings show that successful VLP assembly can be achieved by utilizing recombinant baculoviruses expressing the RSV pre-F or G proteins with the native matrix protein. Mice immunized with either pre-F or the G antigen-expressing VLPs elicited robust serum-mediated virus neutralization. VLP immunization evoked Th1-biased RSV-specific antibody responses in the sera of mice. Following challenge infection with the RSV A2 strain, immunized mice experienced lesser eosinophil and IL-4 accumulation in the lungs, though a substantial increase in TNF-α secretion was observed from CD4+ T cells. Interestingly, splenic antibody-secreting cell responses were substantially enhanced against RSV F antigen, but not against the RSV G antigen following immunization and challenge infection. Immunizing mice with the VLPs significantly inhibited pulmonary histopathology development, as indicated by the diminished inflammatory immune cell influx and mucin secretion. Conclusion: Combined, these vaccine-induced immune responses contributed to successfully inhibiting the RSV replication in the lungs of mice and demonstrated that RSV VLP assembly using insect cell-derived homologous RSV matrix protein is a feasible approach.

Immunogenicity Evaluation of Respiratory Syncytial Virus Prefusogenic-F Based Virus-like-Particles Consisting of G and M Proteins in Mice

“…The reasons for using preFg F protein in our study are (i) preFg does not get converted to the postF conformation, (ii) consists of the p27 protein and (iii) preFg is shown to be more immunogenic than the preF form [31]. Co-infection with two or more rBVs for simultaneous expression of three proteins is shown to give rise to VLPs that are released from cells similar to native RSV [27,36]. Similarly, we employed baculovirus encoding preFg and co-infected the cells with baculoviruses encoding G and M proteins to obtain the VLPs.…”

“…Previously, RSV-VLPs were made using F0 or preF with or without G proteins [33]. More recently, VLPs were developed using M or M2-1 together with preF and G [34][35][36]. These preF-based RSV-VLP were immunogenic and protective.…”