Undergraduate Researcher Cal Poly, Pomona Walnut, California, United States
Introduction/Rationale: West Nile Virus (WNV) is the leading cause of mosquito-borne neuroinvasive disease in the United States. Despite its significant public health impact, no licensed vaccine exists for human use. Dendritic cells (DCs), particularly those expressing the endocytic receptor DEC-205, play a central role in antigen uptake and cross-presentation to T cells, leading to immune activation. To exploit this pathway, a single chain variable fragment (scFv) specific for mouse DEC-205 was engineered and fused to an immunodominant peptide from the NS4b protein of WNV. The scFv–NS4b fusion construct was cloned, expressed in mammalian cells, and purified by affinity chromatography. Binding specificity was confirmed via ELISA and flow cytometry using DEC-205+ cell lines. This work establishes a targeted antigen delivery system for future immunotherapeutic testing. The scFv construct represents a novel tool to selectively direct viral antigens to DCs in vivo, setting the stage for preclinical evaluation in models of neuroinvasive WNV disease.
Methods: We will be using an scFv modeled after ones previously used to interrogate the potential of an aDEC-205 scFv as a therapeutic approach to diseases, including those used by Lakhrif et al. (2018) and Coconi-Linares et al. (2013). This engineered scFv will be genetically conjugated to the immunodominant MHC-class I restricted peptide from West Nile virus. After expressing and purifying the protein in question, we will be using ELISA and a flow cytometry competition assay to assess whether the scFv is indeed binding to DEC-205, an essential step in verifying a specific therapeutic for further downstream testing.
Results: If successful, we expect to see that DEC-205+ cells incubated with the aDEC-205 WNV scFv are able to efficiently block the binding of already-characterized aDEC-205 mAbs.
Conclusion: If our results hold, we will conclude that the scFv protein is an appropriate candidate for testing in a mouse model of neuroinvasive West Nile virus encephalitis.
