- Research note
- Open Access
Functional evaluation for adequacy of MDCK-lineage cells in influenza research
© The Author(s) 2019
- Received: 4 January 2019
- Accepted: 15 February 2019
- Published: 26 February 2019
Influenza is an acute respiratory disease caused by the influenza virus which circulates annually in populations of different species. Madin-Darby Canine Kidney (MDCK) is the most widely utilized cell-line for conducting influenza research. However, the infectivity of various influenza strains in MDCK cells is not equivalent and the productivity of viral propagation is also limited.
We tested the functional adequacy of two MDCK-lineage cell lines, conventional MDCK and MDCK/London, were evaluated by assessing their infectivity of different influenza viral strains with focus forming assays and the cellular toxicity caused by influenza infections by lactate dehydrogenase assay. Moreover, the sensitivity of cells in the presence of the antiviral agent ribavirin was assessed by MTT assay. Our results showed that MDCK/London cells efficiently propagate virus across all influenza viruses tested, are comparable to the utility of Mv1Lu cells, and are superior to conventional MDCK cells in replicating virus as indicated by an increase in virus of three to four logs, particularly in H3N2 infection. Also, the MDCK/London cells were more sensitive to the presence of antiviral drug than conventional MDCK cells. In conclusion, MDCK/London cell line could be a better platform for influenza studies and vaccine development.
- Influenza virus
Influenza virus causes pandemics and seasonal epidemics worldwide, leading to 5–15% of the population becoming infected and 50,000 deaths annually in United States . Frequently complicated by bacterial infection, influenza infection can lead to influenza-associated pneumonia which, in combination, is implicated in up to 8.4% of all reported deaths to the 122 Cities Mortality Reporting System in the United States . Researchers have widely utilized influenza-susceptible cell lines such as MDCK or Vero for pathogenesis investigation as well as vaccine developments, particularly MDCK culture-derived vaccine against influenza has been approved by European Medicines Agency [3, 4]. As such, it is critical to select the cell type utilized for conducting influenza research and to interpret data generated from these various cell types. Currently, several cell lines in addition to MDCK and Vero have been reported to be susceptible to influenza virus infection such as hamster lung (HmLu-1), monkey kidney (JTC-12), human colon intestinal epithelium cell line (CACO-2), and mink lung epithelial cells (Mu1Lv) [5–7].
The MDCK/London (MDCK/Ln) cell line is also a suitable substrate to grow and isolate influenza virus and is available for purchase on the website of Influenza Reagent Resource (IRR; FR-58). In 1985, MDCK/London cells were originated and developed from the Common Cold Laboratory in Salisbury, UK. MDCK/Ln cells display enhanced sensitivity to influenza infections [8, 9]. A recent study revealed that the MDCK/Ln has a faster growth rate and reflects the influenza infection more sensitively in regards to PFU, HA, and NA titers compared to MDCK/SIAT1 and conventional MDCK cells . However, a thorough, in-depth, and functional comparison of MDCK/Ln and conventional MDCK cells has yet to be demonstrated. Here, we compare the viral propagation of MDCK-lineage cells with another cell line, Mv1Lu, which has been shown to be susceptible to influenza infections , by a high throughput focus forming assay which has been described previously .
Next, we performed functional assays in order to further characterize MDCK/Ln cells as a model for influenza infection. We evaluated the cytotoxicity resulting from influenza infections by Lactate dehydrogenase (LDH) assays (CytoTox 96®, Promega). The cytotoxicity, as measured by the increase of LDH, in both MDCK and MDCK/Ln cells continually increased following infection with H1N1, H3N2, and influenza B virus (IBV; ATCC: VR1883). However, compared to the gradual increase of LDH concentration in conventional MDCK cells, the LDH concentration in MDCK/Ln cells increased robustly by Day 2 post-infection and remained at higher levels (Fig. 2c, d).
MDCK cells are widely used in influenza research and vaccine development [17, 18], but the viral yield and efficacy of MDCK-derived vaccines are limited [19, 20]. While further investigation of the underlying mechanisms is required, our functional evaluation for MDCK-lineage cells as a model for influenza infections provide an alternative aspect of MDCK/Ln cells. We believe that our research findings regarding MDCK/Ln cells can be used to further the flu vaccine development and influenza-related research.
HCT led, drafted and performed the experiments in this study. CWL and SWT collected necessary samples and interpreted the data. CCL performed quantification of images and contributed majorly in manuscript writing and CMC analyzed and evaluated the data in this study for publication in addition to acquire funding management and supervised the whole study. All authors read and approved the final manuscript.
We are grateful for the valuable opinions from Dr. Mathew Matt at Booz Allen Hamilton to improve this manuscript.
The authors declare that they have no competing interests.
Availability of data and materials
All data generated or analyzed during this study are included in this published article.
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Ethics approval and consent to participate
This study was internally funded by Taichung Hospital and National Chung Hsing University (MOE-108-S-0023) in terms of providing experimental supplies and subsidizing labor cost. The funder has no influence on experimental design, data interpretation or manuscript writing, editing, approval, or decision to publish.
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