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Liang, Yinjie; Guo, Junjia; Li, Zhen; Liu, Shiyuan; Zhang, Ting; Sun, Shucai; Lu, Funa; Zhai, Yuqian; Wang, Wenling; Ning, Chuanyi; Tan, Wenjie
Biosafety and health, June 2024, Volume: 6, Issue: 3Journal Article
•Scientific question: Low expression of matrix protein 2 (M2) in cells infected with the influenza virus hampers studies.•Evidence before this study: The protective role of influenza virus M2 through antibody-dependent cell-mediated cytotoxicity (ADCC) is demonstrated. Inducing ADCC can effectively clear virus-infected cells, serving as a vital bridge between humoral and cellular immunity, as previous studies have suggested. Therefore, the selection of candidate vaccines capable of eliciting both broad neutralizing antibodies and potent ADCC is crucial for controlling influenza A virus outbreaks.•New findings: A stable cell line (YAC-1-M2) capable of sustaining M2 expression was established. The optimal effector-to-target cell ratio for this target cell is 20:1, and the IgG2a subtype is more effective in inducing ADCC compared to the IgG1 subtype.•Significance of the study: This study facilitates comprehension of the immunoprotective mechanisms associated with M2 and lays the groundwork for the development of vaccines and drugs based on non-neutralizing antibody protection mechanisms. The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity (ADCC) associated with M2-based immunization in mice. We first established a stable cell line derived from mouse lymphoma cells (YAC-1) expressing M2 of H3N2. This cell line, designated as YAC-1-M2, was generated using a second-generation lentiviral tricistronic plasmid system to transduce the M2 gene into YAC-1 cells. The ADCC effect induced by polyclonal antibodies targeting matrix protein 2 ectodomain (M2e) was demonstrated by YAC-1-M2 cell lysis by natural killer cells (NK) derived from mice, in the presence of anti-M2 antibodies obtained from mice immunized with an mRNA vaccine based on M2e. This ADCC effect was found to be stronger compared to the effect induced by monoclonal antibodies (14C2) against M2. Moreover, the ADCC effect was enhanced as the effector-to-target ratio of NK to YAC-1-M2 cells increased. In conclusion, we established a novel method to detect ADCC of M2 of IAV, which paves the way for the development of an M2-based universal vaccine against IAV and an in-depth analysis of its mechanism of broad-spectrum immune protection in mice.
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