Growth and production of the dengue virus in C6/36 cells and identification of a laminin-binding protein as a candidate serotype 3 and 4 receptor protein

Objectives: Although dengue is one of the most common mosquito-borne viral diseases, few studies have investigated the relationship between the dengue virus and mosquito cells, and this study sought to describe the binding and propagation of the dengue viruses in C6/36 cells. Methods: The internalization and production of the dengue virus was assayed by standard plaque assay methodologies, while dengue virus receptor proteins were examined by a virus overlay protein-binding assay and candidate gene analysis coupled with virus inhibition studies. Results: All four serotypes were internalized linearly, and de novo virus production occurred 14-19 h postinfection. Virus overlay protein-binding assay identified a band of 50 kDa for dengue serotypes 2, 3 and 4 which comigrated with a protein that reacts with anti-bodies directed against the human 37/67-kDa high-affinity laminin receptor. Both antibodies directed against the human 37/67-kDa high-affinity laminin receptor protein and soluble laminin inhibited the binding and internalization of serotypes 3 and 4, but not serotypes 1 and 2. Conclusions: The results suggest that multiple receptors may be used by the dengue virus to enter into insect cells, and that one of these proteins may be a laminin-binding protein.