✍🏼 Xuemin Chen, He-ying Sun, Chun Yi Lee, Christina A. Rostad, Jessica Trost, Rodrigo B. Abreu, Michael A. Carlock, Jason R. Wilson, Shane Gansebom, Ted M. Ross, David A. Steinhauer, Evan J. Anderson, Larry J. Anderson
🏠 Division of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, United States
📑 Virology (2022)
Read the Article
Abstract
Novel cell-based assays were developed to assess antibody-dependence cellular cytotoxicity (ADCC) antibodies against both vaccine and a representative circulation strain HA and NA proteins for the 2014-15 influenza season. The four assays using target cells stably expressing one of the four proteins worked well. In pre- and post-vaccine sera from 70 participants in a pre-season vaccine trial, we found ADCC antibodies and a rise in ADCC antibody titer against target cells expressing the 4 proteins but a much higher titer for the vaccine than the circulating HA in both pre-and post-vaccine sera. These differences in HA ADCC antibodies were not reflected in differences in HA binding antibodies. Our observations suggested that relatively minor changes on the subtype HA can result in large differences in ADCC activity.
How is the WOLF used in this study
The WOLF cell sorter was used to ensure the clonality of hybridoma cells producing monoclonal antibodies against influenza virus antigens. After initial hybridoma generation and screening, positive antibody-producing clones were subjected to single-cell sorting using the WOLF, which provided gentle, microfluidic fluorescence-activated cell sorting to isolate individual hybridoma cells into separate wells. This allowed the researchers to derive stable monoclonal cell lines from single founder cells, which is critical for consistent antibody production and for characterizing functional ADCC activity of the resulting antibodies.
