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Use of ADCC Reporter Bioassays in Influenza Vaccine Development

Harmjan Kuipers did his PhD in immunology in the lab of Prof. Lambrecht in Rotterdam, The Netherlands focusing on dendritic cell based immunotherapy in murine models of asthma. He then moved for a 2-year postdoc project to the lab of Prof. Brocker in Munich, studying mircoRNAs in dendritic cell development and function. In 2008 he started to work at Crucell, now part of Janssen Vaccines, where is he is currently involved in the development of a universal influenza vaccine.
  • Dr. Harmjan Kuipers, Janssen Pharmaceutical, Infectious Diseases and Vaccines

  • Sr. Scientist Vaccine Discovery and Early Development

  • Original Webinar Date: Tuesday, May 10, 2016

This webinar will discuss the application of Promega's ADCC reporter bioassays in influenza vaccine development with special emphasis on the setup and use and of a murine FcγRIV version of this assay.

The identification of human broadly neutralizing antibodies (bnAbs) targeting the hemagglutinin (HA) stem revitalized hopes of developing a universal influenza vaccine. Using a rational design and library approach, we engineered and produced stable HA stem antigens (‘mini-HAs’). Our most advanced candidate exhibited structural and bnAb binding properties comparable to full-length HA.  We observed complete protection in stringent heterologous and heterosubtypic lethal influenza challenge mouse models and also demonstrated that protection is serum-mediated. In Cynomolgus monkeys, this mini-HA reduced fever following sublethal H1N1 challenge. Antibodies elicited by this mini-HA in mice and monkeys bound a wide range of HAs, competed with human bnAbs for HA stem binding and neutralized H5N1 viruses. It has become increasingly clear that an important in vivo effector function of bnAbs targeting the HA stem consists of antibody-dependent effector mechanism such antibody-dependent cellular cytotoxicity (ADCC). Using ADCC reporter bioassays we showed that sera from mini-HA immunized animals have indeed ADCC activity. To measure antibody-dependent effector mechanisms in murine samples, we used a recently developed variant ADCC reporter bioassay of the human FcγRIIIa assay, based on murine FcγRIV. The second part of the webinar will focus on setup of this assay, analyzing the effect of varying key assay parameters, specificity of the assay and considerations when using serum samples instead of monoclonal antibodies.