Nipaporn KanthongNuanpan KhemnuSa Nga PattanakitsakulPrida MalasitTimothy W. FlegelRajamangala University of Technology Tawan-okMahidol UniversityThailand National Center for Genetic Engineering and BiotechnologyThailand National Science and Technology Development Agency2018-09-242018-09-242010-02-22BMC Microbiology. Vol.10, (2010)14712180147121802-s2.0-76749138603https://repository.li.mahidol.ac.th/handle/20.500.14594/29270Background. It is known that insects and crustaceans can carry simultaneous, active infections of two or more viruses without showing signs of disease, but it was not clear whether co-infecting viruses occupied the same cells or different cells in common target tissues. Our previous work showed that successive challenge of mosquito cell cultures followed by serial, split-passage resulted in stabilized cultures with 100% of the cells co-infected with Dengue virus (DEN) and an insect parvovirus (densovirus) (DNV). By addition of Japanese encephalitis virus (JE), we tested our hypothesis that stable, persistent, triple-virus co-infections could be obtained by the same process. Results. Using immunocytochemistry by confocal microscopy, we found that JE super-challenge of cells dually infected with DEN and DNV resulted in stable cultures without signs of cytopathology, and with 99% of the cells producing antigens of the 3 viruses. Location of antigens for all 3 viruses in the triple co-infections was dominant in the cell nuclei. Except for DNV, this differed from the distribution in cells persistently infected with the individual viruses or co-infected with DNV and DEN. The dependence of viral antigen distribution on single infection or co-infection status suggested that host cells underwent an adaptive process to accommodate 2 or more viruses. Conclusions. Individual mosquito cells can accommodate at least 3 viruses simultaneously in an adaptive manner. The phenomenon provides an opportunity for genetic exchange between diverse viruses and it may have important medical and veterinary implications for arboviruses. © 2010 Kanthong et al; licensee BioMed Central Ltd.Mahidol UniversityImmunology and MicrobiologyMedicineArticleSCOPUS10.1186/1471-2180-10-14