Problem: Increased disease severity with sequential infections may be due to antibody dependent enhancement. Macrophages do not have the normal dengue virus receptor. However, incomplete neutralization of virus can target virions to macrophage Fc receptors.
Q: How can we find out where neutralizing antibodies target the viral envelope protein, so we can use just these areas in a vaccine? A: Characterize the binding targets (epitopes) for individual human antibodies. (subquestion 1: how in the world do you get individual human antibodies???)
Immortalize B-cells with EBV and clone by serial dilution. Screen for binding to DENV. Characterize binding, neutralization, enhancement, and epitope mapping. We’re going to need IRB approval…
Ethical questions: Will people be hurt by this procedure? How can we minimize this risk? What will the fate of the cells be?
Different collaborating institutions: FGCU Tulane University Tan Tock Seng Hospital University of the West Indies
Different procedures. Different approval process. Different regulations. Different consent forms. Who even approves first?
Two full FGCU IRB applications with one amendment. All samples blinded and coded. Samples from Florida, Jamaica, and Singapore. No adverse incidences. Project ongoing, multiple HuMAbs isolated and characterized. First description of HuMAbs against dengue virus.
Neutralizing and non-neutralizing monoclonal antibodies against dengue virus E protein derived from a naturally infected patient Virology Journal 2010, 7:28 doi:10.1186/1743-422X-7-28 John S Schieffelin (firstname.lastname@example.org) Joshua M Costin (email@example.com) Cindo O Nicholson (firstname.lastname@example.org) Nicole M Orgeron (email@example.com) Krystal A Fontaine (firstname.lastname@example.org) Sharon Isern (email@example.com) Scott F Michael (firstname.lastname@example.org) James E Robinson (email@example.com)
FGCU Sharon Isern, PhD Joshua Costin, PhD Kelli Barr, PhD Yancey Hrobowski, PhD Krystal Fontaine Cindo Nicholson Craig Rees Tom Everts Nadiya Joseph Collaborators Li Lin, MD - Tan Tock Seng John Lindo, MD - UWI Mona James Robinson, MD - Tulane John Schefflein, MD - Tulane Funding US National Institutes of Health US Department of Defense Acknowledgements