Journal of Microbiology

Research article

Development and analytical evaluation of a microneutralization cytopathic effect assay for human adenovirus type 55-specific neutralizing antibodies: Dae-Im Jung, Yunjeong Park, Jung-ah Choi, Soon-Hwan Kwon, Jun Young Lee, Manki Song, Sang Hwan Seo; J. Microbiol. 2026;64(7):e2604007. Published online July 6, 2026; DOI: https://doi.org/10.71150/jm.2604007

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Abstract PDF Supplementary Material: Reliable quantification of neutralizing antibodies (nAb) against human adenovirus type 55 (HAdV-55) is critical for the evaluation of emerging vaccine candidates. While the plaque reduction neutralization test (PRNT) is currently the reference standard, its utility for large-scale studies is limited by low throughput, labor-intensive plaque counting, and prolonged assay times. In this study, we established and analytically validated a microneutralization assay based on cytopathic effect (MN-CPE) as a scalable alternative for HAdV-55-specific nAb quantification. Comparative performance analysis revealed that both assays maintain high dilution linearity, with coefficients of determination (R²) of 0.988 for MN-CPE and 0.9926 for PRNT. Relative accuracy assessments using high-, middle-, and low-titer reference sera demonstrated acceptable responses across the dynamic range. Notably, the MN-CPE assay allowed for the definition of a negative-control acceptance range, providing a distinct statistical advantage over PRNT, where negative-control values were consistently zero. Furthermore, both assays successfully detected HAdV-55-specific nAbs in immunized cynomolgus macaques, with no cross-reactivity observed against other HAdV types such as HAdV-4. These findings indicate that the MN-CPE assay is analytically comparable to PRNT and serves as a practical, relatively high-capacity alternative for HAdV-55 neutralization testing in clinical and preclinical vaccine research.

Article

Mouse strain-dependent neutralizing antibody responses to Zika virus vaccines: Sang Hwan Seo, Jung-ah Choi, Eunji Yang, Hayan Park, Dae-Im Jung, Jae-Ouk Kim, Jae Seung Yang, Manki Song; J. Microbiol. 2025;63(8):e2504005. Published online August 31, 2025; DOI: https://doi.org/10.71150/jm.2504005

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The 2015 Zika virus (ZIKV) outbreak in Brazil and its global spread underscored the urgent need for effective and broadly protective vaccines. While C57BL/6 and BALB/c mice are widely used in preclinical vaccine research, direct comparisons of their ability to elicit ZIKV-specific neutralizing antibodies (nAbs) remain limited. This study aimed to systematically evaluate and compare the immunogenic potential of these two common mouse strains across diverse vaccine platforms, focusing on their capacity to generate functional neutralizing antibody responses. We assessed nAb and IgG responses following four vaccination strategies: (1) DNA vaccine encoding prMEΔTM followed by E protein domain III boost, (2) recombinant EΔTM protein expressed using baculovirus system, (3) formalin-inactivated ZIKV, and (4) live ZIKV. Although both strains generated detectable ZIKV- and E protein-specific IgG, the magnitude and quality of responses varied by vaccine platform and strain. Notably, C57BL/6 mice consistently mounted significantly higher nAb titers than BALB/c mice across all immunization groups, including subunit- and whole-virus-based vaccines. In contrast, BALB/c mice showed lower or undetectable nAb responses, despite comparable or higher total IgG levels in some cases. These findings show that host genetic background is a critical determinant of vaccine-induced neutralization and underscore the importance of selecting appropriate animal models in ZIKV vaccine development. C57BL/6 mice, due to their robust nAb responses, represent a reliable model for evaluating vaccine immunogenicity. Conversely, the limited nAb responses in BALB/c mice position them as a potential low-responder model, offering a stringent system to test the potency and breadth of protective immunity under suboptimal conditions.

Citations

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The Pathogenesis and Virulence of the Major Enterovirus Pathogens Associated with Severe Clinical Manifestations: A Comprehensive Review
Yuwei Liu, Maiheliya Maisimu, Zhihang Ge, Suling Xiao, Haoran Wang
Cells.2025; 14(20): 1617. CrossRef
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Validation Study

Comparison of JEV Neutralization Assay Using Pseudotyped JEV with the Conventional Plaque-Reduction Neutralization Test: Hee-Jung Lee , Kyung-Il Min , Ki Hoon Park , Hyo Jung Choi , Min-Kyoung Kim , Chi-Young Ahn , Young-Jin Hong , Young Bong Kim; J. Microbiol. 2014;52(5):435-440. Published online March 7, 2014; DOI: https://doi.org/10.1007/s12275-014-3529-y

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Abstract PDF

We previously reported the development of a neutralization assay system for evaluating Japanese Encephalitis Virus (JEV) neutralizing antibody (NAb) using pseudotyped-JEV (JEV- PV). JEV-PV-based neutralization assay offers several advan-tages compared with the current standard plaque-reduc-tion neutralization test (PRNT), including simplicity, safety, and speed. To evaluate the suitability of the JEV-PV assay as new replacement neutralization assay, we compared its repeatability, reproducibility, specificity, and correlated its results with those obtained using the PRNT. These analyses showed a close correlation between the results obtained with the JEV-PV assay and the PRNT, using the 50% plaque re-duction method as a standard for measuring NAb titers to JEV. The validation results met all analytical acceptance criteria. These results suggest that the JEV-PV assay could serve as a safe and simple method for measuring NAb titer against JEV and could be used as an alternative approach for assaying the potency of JEV neutralization.

Citations

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Role of pseudotyped viruses in understanding epidemiology, pathogenesis and immunity of viral diseases affecting both horses and humans
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Frontiers in Immunology.2023;[Epub] CrossRef
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Byung Ok Kwak, Young Jin Hong, Dong Hyun Kim
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Journal of Microbiology.2020; 58(1): 46. CrossRef
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Hee-Jung Lee, Hanul Choi, Ki Hoon Park, Yuyeon Jang, Young-jin Hong, Young Bong Kim
International Journal of Infectious Diseases.2020; 100: 53. CrossRef
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Scientific Reports.2020;[Epub] CrossRef
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Byung Ok Kwak, Young Se Kwon, Young Jin Hong, Chung Hyun Nahm, Woori Jang, Young Uh, Yong Gon Cho, Jimyung Kim, Myungshin Kim, Dong Hyun Kim
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Journal of Immunology Research.2019; 2019: 1. CrossRef
MLKL Mediated Necroptosis Accelerates JEV-Induced Neuroinflammation in Mice
Peiyu Bian, Xuyang Zheng, Li Wei, Chuantao Ye, Hong Fan, Yanhui Cai, Ying Zhang, Fanglin Zhang, Zhansheng Jia, Yingfeng Lei
Frontiers in Microbiology.2017;[Epub] CrossRef
Carbon nanoparticle modified screen printed carbon electrode as a disposable electrochemical immunosensor strip for the detection of Japanese encephalitis virus
Suk Fun Chin, Lih Shan Lim, Suh Cem Pang, Magdline Sia Henry Sum, David Perera
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