Pseudovirus-Based Strategies for Understanding and Controlling Viral Infections

A special issue of Pathogens (ISSN 2076-0817). This special issue belongs to the section "Viral Pathogens".

Deadline for manuscript submissions: 31 October 2026 | Viewed by 3905

Special Issue Editor


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Guest Editor
Division of HIV/AIDS and Sex-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing 102629, China
Interests: SARS-CoV-2; HIV-1; HPV; vaccine development; vaccine evaluation; immune response; neutralizing antibody; standardization of assay
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Special Issue Information

Dear Colleagues,

Pseudoviruses have become indispensable tools in biomedical research, serving as vital surrogates for authentic viruses across multiple applications. In broad terms, these recombinant constructs are defined by their incorporation of foreign genetic material, regardless of whether they display exogenous surface proteins. This versatile category has application in understanding the infectivity mechanisms, gene therapy delivery systems, vaccine vector development, and diagnostic control materials for nucleic acid assays.

Despite significant progress in pseudovirus development for emerging pathogens, substantial technical challenges persist. Current limitations include a lack of universal construction strategies, persistent technical barriers for certain virus families (e.g., flaviviruses), and inconsistent standardization across research groups.

Hence, this Special Issue, titled “Pseudovirus-Based Strategies for Understanding and Controlling Viral Infections”, promotes studies that focus on advanced pseudovirus packaging strategies; their use in studies of virus life cycles; emerging applications in vaccines, therapeutics, and diagnostics; and any other fields related to pseudovirses.

Dr. Jianhui Nie
Guest Editors

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Keywords

  • pseudovirus
  • infectivity
  • neutralization
  • antigenicity
  • immunogenicity

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Published Papers (2 papers)

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Research

21 pages, 3748 KB  
Article
Pseudovirus-Based Neutralization Assays as Customizable and Scalable Tools for Serological Surveillance and Immune Profiling
by Caio Bidueira Denani, Bruno Pimenta Setatino, Denise Pereira, Ingrid Siciliano Horbach, Adriana Souza Azevedo, Gabriela Coutinho, Clara Lucy Ferroco, Janaína Xavier, Robson Leite, Ewerton Santos, Maria de Lourdes Maia, Waleska Dias Schwarcz and Ivanildo Pedro Sousa
Pathogens 2025, 14(11), 1129; https://doi.org/10.3390/pathogens14111129 - 6 Nov 2025
Cited by 3 | Viewed by 2212 | Correction
Abstract
Neutralizing antibodies (nAbs) are key indicators of protection against SARS-CoV-2, and their measurement remains essential for monitoring vaccine responses and population immunity. While the plaque reduction neutralization test (PRNT) is the gold standard, it relies on replicative viruses and is not suited for [...] Read more.
Neutralizing antibodies (nAbs) are key indicators of protection against SARS-CoV-2, and their measurement remains essential for monitoring vaccine responses and population immunity. While the plaque reduction neutralization test (PRNT) is the gold standard, it relies on replicative viruses and is not suited for high-throughput applications. Here, both an in-house and a commercial pseudovirus-based neutralization (PBN) assay were standardized and compared with PRNT to assess performance and concordance. The in-house PBN employed a VSV-ΔG pseudovirus encoding NanoLuc and displaying the SARS-CoV-2 Spike from the Wuhan or Omicron BA.1 variants in HEK293T-hACE2 cells, whereas the commercial assay (Integral Molecular, Philadelphia, PA, USA) used a lentiviral backbone with Renilla or GFP reporters and Wuhan or Omicron XBB.1.5/XBB.1.9 Spikes in Vero E6-ACE2-TMPRSS2 cells. Both assays showed strong correlations with PRNT, the commercial assay; moreover, they offered superior reproducibility and scalability, while the in-house version provided a cost-effective alternative suitable for BSL-2 settings. A total of 600 serum samples from vaccinated individuals were analyzed by commercial PBN at collection time points, from pre-vaccination to twelve months post–second dose, enabling large-scale screening, revealing marked differences in neutralization between Wuhan and Omicron XBB.1.5/1.9, and allowing unbiased classification of low, medium, and high responders using k-means clustering. The geometric mean titers (log10 GMT) highlighted a ~1.5 log10 (eightfold) reduction in neutralizing activity against Omicron, reflecting antibody waning and antigenic drift. Altogether, this study integrates assay standardization, PRNT comparison, and large-scale immune profiling, establishing a robust framework for harmonized pseudovirus-based neutralization testing. Full article
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12 pages, 4204 KB  
Article
Establishment of a Pseudovirus-Based Golden Hamster Model for the Attachment and Entry Stages of Hendra Virus Infection and Evaluation of Protective Immunity
by Tao Li, Binfan Liao, Danfeng Li, Jie Zhang, Chunhui Zhao, Yunfei Pei, Liping Chen, Meng Wang, Yawen Liu, Xi Wu, Weijin Huang and Jianhui Nie
Pathogens 2025, 14(9), 910; https://doi.org/10.3390/pathogens14090910 - 10 Sep 2025
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Abstract
Objective: Establish an in vivo evaluation model focused on the attachment and entry stages of Hendra virus infection for protective immunity assessment. Methods: A golden hamster infection model based on recombinant Hendra-F/G pseudovirus was developed, and a luciferase luminescence assay was used to [...] Read more.
Objective: Establish an in vivo evaluation model focused on the attachment and entry stages of Hendra virus infection for protective immunity assessment. Methods: A golden hamster infection model based on recombinant Hendra-F/G pseudovirus was developed, and a luciferase luminescence assay was used to assess the optimal pseudoviral challenge in terms of route of infection, dose and detection time. The biodistribution of the pseudovirus in infected organs was evaluated using the IVIS spectral CT system. The protective effect of antibody prophylaxis was evaluated by measuring the luminescence intensity of pseudoviruses. Results: Intraperitoneal injection was identified as the optimal route of infection, and the optimal time of detection was 6 h post-challenge. Our model simulates the infection of the brain and lungs by live viruses, with the strongest infection occurring in the abdomen, especially in the intestinal organs. The dose of pseudovirus was linearly correlated with luminescence intensity. The infection model was able to differentiate the protective effect of monoclonal antibodies, with complete protection in the high-dose group. Conclusions: The recombinant Hendra-F/G pseudovirus hamster model allows the effective evaluation of prophylactic monoclonal antibodies, providing a crucial tool for studying Hendra virus infection and control strategies. Full article
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