2026-04-27T18:25:44Zhttps://riubu.ubu.es/oai/request

oai:riubu.ubu.es:10259/109202025-10-04T00:05:21Zcom_10259_4862com_10259_5086com_10259_2604col_10259_4863

Activity of human serum antibodies in an influenza virus hemagglutinin stalk-based ADCC reporter assay correlates with activity in a CD107a degranulation assay Chromikova, Veronika Tan, Jessica Aslam, Sadaf Rajabhathor, Arvind Bermudez-Gonzalez, Maria Ayllón Barasoain, Juan Simon, Viviana García Sastre, Adolfo Salaun, Bruno Nachbagauer, Raffael Krammer, Florian Influenza hemagglutinin ADCC ADCP Effector functions Stalk antibodies Seroterapia Serotherapy The stalk of the influenza virus hemagglutinin (HA) is an attractive target for antibody-based universal influenza virus vaccine development. While antibodies that target this part of the virus can be neutralizing, it has been shown in recent years that Fc receptor-mediated effector functions are of significant importance for the protective effect of anti-stalk antibodies. Several assays to measure Fc-Fc receptor interaction-based effector functions like antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis exist, but they suffer from limitations such as low throughput and high run-to-run variability. Reporter assays for antibody-dependent cellular cytotoxicity based on reporter cells that express luciferase upon engagement of human FcγRIIIa with the Fc of antigen-bound antibodies have been developed as well. These reporter assays can be used in a higher throughput setting with limited run-to-run assay variability but since they express only one Fc receptor, their biological relevance is unclear. Here we optimized an antibody-dependent cellular cytotoxicity reporter assay to measure the activity of antibodies to the conserved stalk domain of H1 hemagglutinin. The assay was then correlated to a CD107a-based degranulation assay, and a strong and significant correlation could be observed. This data suggests that the FcγRIIIa-based reporter assay is a good substitute for functional assays, especially in settings where larger sample numbers need to be analyzed. Work in the Krammer laboratory was supported by GlaxoSmithKline as well as funding from the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS, HHSN272201400008C). We would like to thank Dr. Patrick Wilson from the University of Chicago for providing plasmids for human mAb expression, the Icahn School of Medicine Flow Cytometry Core for access to the flow cytometers and Dr. Evren Azeloglu for assistance with fluorescence microscopy. We would also like to thank Elena Hirsch for expert sample processing as well as the study participants. 2025-10-03T11:16:48Z 2025-10-03T11:16:48Z 2020-02 info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion 0264-410X https://hdl.handle.net/10259/10920 10.1016/j.vaccine.2020.01.008 1873-2518 eng Vaccine. 2020, V. 8, n. 38, p.1953–1961 https://doi.org/10.1016/j.vaccine.2020.01.008 Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess application/pdf Elsevier https://riubu.ubu.es/bitstream/10259/10920/5/Chromikova_Vaccine_2020.pdf.jpg Hispana TEXT http://creativecommons.org/licenses/by-nc-nd/4.0/ RIUBU. Repositorio Institucional de la Universidad de Burgos https://hdl.handle.net/10259/10920