tetano
Editor, Senior Moderator
Vaccine. 2020 Jan 17. pii: S0264-410X(20)30012-8. doi: 10.1016/j.vaccine.2020.01.008. [Epub ahead of print] [h=1]Activity of human serum antibodies in an influenza virus hemagglutinin stalk-based ADCC reporter assay correlates with activity in a CD107a degranulation assay.[/h]
Chromikova V[SUP]1[/SUP], Tan J[SUP]2[/SUP], Aslam S[SUP]1[/SUP], Rajabhathor A[SUP]1[/SUP], Bermudez-Gonzalez M[SUP]1[/SUP], Ayllon J[SUP]1[/SUP], Simon V[SUP]1[/SUP], Garc?a-Sastre A[SUP]3[/SUP], Salaun B[SUP]4[/SUP], Nachbagauer R[SUP]1[/SUP], Krammer F[SUP]5[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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.
Copyright ? 2020. Published by Elsevier Ltd.
[h=4]KEYWORDS:[/h] ADCC; ADCP; Effector functions; Influenza hemagglutinin; Stalk antibodies
PMID: 31959425 DOI: 10.1016/j.vaccine.2020.01.008
Chromikova V[SUP]1[/SUP], Tan J[SUP]2[/SUP], Aslam S[SUP]1[/SUP], Rajabhathor A[SUP]1[/SUP], Bermudez-Gonzalez M[SUP]1[/SUP], Ayllon J[SUP]1[/SUP], Simon V[SUP]1[/SUP], Garc?a-Sastre A[SUP]3[/SUP], Salaun B[SUP]4[/SUP], Nachbagauer R[SUP]1[/SUP], Krammer F[SUP]5[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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.
Copyright ? 2020. Published by Elsevier Ltd.
[h=4]KEYWORDS:[/h] ADCC; ADCP; Effector functions; Influenza hemagglutinin; Stalk antibodies
PMID: 31959425 DOI: 10.1016/j.vaccine.2020.01.008