Vaccine · 2026
AEX-1: A Vero cell variant with enhanced permissiveness for influenza virus replication.
СтатьяНазвание и аннотация: Перевод пока не подготовлен
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Krokhin Artem, Romanova Julia, Grillari Regina, Hackl Matthias, Korabliov Pavel, Mandík Jozef, Egorov Andrej
Аннотация
BACKGROUND: Continuous cell substrates with improved permissiveness for influenza virus replication are of considerable interest for vaccine development and manufacturing. Vero cells have an excellent regulatory track record; however, their relatively low influenza virus productivity limits their broader application.
METHODS: We developed the AEX-1 cell line by co-cultivating Vero cells with chicken embryo amniotic membrane fragments. Cellular morphology, intracellular acidification, influenza virus replication, transcriptomic profile, genetic stability, and tumorigenicity were evaluated.
RESULTS: AEX-1 cells displayed distinct morphology and enhanced endosomal/lysosomal acidification compared with parental Vero cells. At 72 h post-infection, a commonly used harvest time for influenza vaccine virus production, AEX-1 cells produced higher infectious titres of WHO-recommended egg-derived A/Sydney/5/2021 (H1N1)pdm09-like and B/Austria/1359417/2021 vaccine viruses, with titres approaching those obtained in embryonated chicken eggs and exceeding Vero cell yields by 0.9-1.7 log10 EID50/ml. No replication advantage was observed for the egg-derived A/Croatia/10136RV/2023 (H3N2) vaccine virus, carrying the egg-adaptive HA G53D substitution, or for the egg-derived A/duck/Vietnam/NCVD-1584/2012 (H5N1) vaccine strain. However, the A/Darwin/9/2021-like Vero-derived primary isolate A/Austria/1516645/2022 (H3N2), lacking G53D, reached infectious titres up to 3.0 log10 TCID50/ml higher than in Vero and 2.3 log10 TCID50/ml higher than in MDCK cells. H1N1 UniFluVec-based influenza virus vectors also replicated efficiently in AEX-1 cells. Transcriptomic analysis revealed changes in genes involved in proton transport, vesicular acidification, and intracellular pH regulation. AEX-1 remained genetically stable after prolonged passaging and showed no evidence of increased tumorigenicity in vitro or in vivo.
CONCLUSIONS: Enhanced intracellular acidification may contribute to improved influenza virus replication in AEX-1 cells, suggesting that modulation of endosomal acidification may provide a rational strategy for developing improved cell substrates for influenza vaccine production.
METHODS: We developed the AEX-1 cell line by co-cultivating Vero cells with chicken embryo amniotic membrane fragments. Cellular morphology, intracellular acidification, influenza virus replication, transcriptomic profile, genetic stability, and tumorigenicity were evaluated.
RESULTS: AEX-1 cells displayed distinct morphology and enhanced endosomal/lysosomal acidification compared with parental Vero cells. At 72 h post-infection, a commonly used harvest time for influenza vaccine virus production, AEX-1 cells produced higher infectious titres of WHO-recommended egg-derived A/Sydney/5/2021 (H1N1)pdm09-like and B/Austria/1359417/2021 vaccine viruses, with titres approaching those obtained in embryonated chicken eggs and exceeding Vero cell yields by 0.9-1.7 log10 EID50/ml. No replication advantage was observed for the egg-derived A/Croatia/10136RV/2023 (H3N2) vaccine virus, carrying the egg-adaptive HA G53D substitution, or for the egg-derived A/duck/Vietnam/NCVD-1584/2012 (H5N1) vaccine strain. However, the A/Darwin/9/2021-like Vero-derived primary isolate A/Austria/1516645/2022 (H3N2), lacking G53D, reached infectious titres up to 3.0 log10 TCID50/ml higher than in Vero and 2.3 log10 TCID50/ml higher than in MDCK cells. H1N1 UniFluVec-based influenza virus vectors also replicated efficiently in AEX-1 cells. Transcriptomic analysis revealed changes in genes involved in proton transport, vesicular acidification, and intracellular pH regulation. AEX-1 remained genetically stable after prolonged passaging and showed no evidence of increased tumorigenicity in vitro or in vivo.
CONCLUSIONS: Enhanced intracellular acidification may contribute to improved influenza virus replication in AEX-1 cells, suggesting that modulation of endosomal acidification may provide a rational strategy for developing improved cell substrates for influenza vaccine production.