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Dengue human infection studies present an opportunity to address many longstanding questions in the field of flavivirus biology. However, limited data are available on how the immunological and transcriptional response elicited by an attenuated challenge virus compares to that associated with a wild-type DENV infection. To determine the kinetic transcriptional signature associated with experimental primary DENV-1 infection and to assess how closely this profile correlates with the transcriptional signature accompanying natural primary DENV-1 infection, we utilized scRNAseq to analyze PBMC from individuals enrolled in a DENV-1 human challenge study and from individuals experiencing a natural primary DENV-1 infection. While both experimental and natural primary DENV-1 infection resulted in overlapping patterns of inflammatory gene upregulation, natural primary DENV-1 infection was accompanied with a more pronounced suppression in gene products associated with protein translation and mitochondrial function, principally in monocytes. This suggests that the immune response elicited by experimental and natural primary DENV infection are similar, but that natural primary DENV-1 infection has a more pronounced impact on basic cellular processes to induce a multi-layered anti-viral state., Author summary Dengue Human Challenge Models allow for the analysis of host/virus interactions under highly controlled experimental conditions. However, it is unclear how close the immune response generated by an attenuated challenge virus compares to that generated by a naturally acquired DENV infection. In this study, we utilized single cell RNA sequencing to assess the immune response generated by both experimental and natural primary DENV-1 infections. This analysis suggests that the immune response elicited by experiential and natural primary DENV-1 infections are similar, but that natural DENV-1 infection has a more pronounced impact on basic cellular processes to induce a multi-layered anti-viral state.

Genetic reassortment between influenza A viruses (IAVs) facilitate emergence of pandemic strains, and swine are proposed as a “mixing vessel” for generating reassortants of avian and mammalian IAVs that could be of risk to mammals, including humans. However, how a transmissible reassortant emerges in swine are not well understood. Genomic analyses of 571 isolates recovered from nasal wash samples and respiratory tract tissues of a group of co-housed pigs (influenza-seronegative, avian H1N1 IAV–infected, and swine H3N2 IAV–infected pigs) identified 30 distinct genotypes of reassortants. Viruses recovered from lower respiratory tract tissues had the largest genomic diversity, and those recovered from turbinates and nasal wash fluids had the least. Reassortants from lower respiratory tracts had the largest variations in growth kinetics in respiratory tract epithelial cells, and the cold temperature in swine nasal cells seemed to select the type of reassortant viruses shed by the pigs. One reassortant in nasal wash samples was consistently identified in upper, middle, and lower respiratory tract tissues, and it was confirmed to be transmitted efficiently between pigs. Study findings suggest that, during mixed infections of avian and swine IAVs, genetic reassortments are likely to occur in the lower respiratory track, and tissue tropism is an important factor selecting for a transmissible reassortant., Author summary Genetic reassortments between avian and swine influenza viruses are likely to occur in the swine lower respiratory track, and tissue tropism is an important factor selecting for a transmissible reassortant; determination of tissue tropisms for potential reassortants between contemporary avian and swine influenza viruses would help identify transmissible reassortants with public health risks.

Studies have demonstrated cross-reactivity of anti-dengue virus (DENV) antibodies in human sera against Zika virus (ZIKV), promoting increased ZIKV infection in vitro. However, the correlation between in vitro and in vivo findings is not well characterized. Thus, we evaluated the impact of heterotypic flavivirus immunity on ZIKV titers in biofluids of rhesus macaques. Animals previously infected (≥420 days) with DENV2, DENV4, or yellow fever virus were compared to flavivirus-naïve animals following infection with a Brazilian ZIKV strain. Sera from DENV-immune macaques demonstrated cross-reactivity with ZIKV by antibody-binding and neutralization assays prior to ZIKV infection, and promoted increased ZIKV infection in cell culture assays. Despite these findings, no significant differences between flavivirus-naïve and immune animals were observed in viral titers, neutralizing antibody levels, or immune cell kinetics following ZIKV infection. These results indicate that prior infection with heterologous flaviviruses neither conferred protection nor increased observed ZIKV titers in this non-human primate ZIKV infection model., Author summary Zika virus (ZIKV) is a mosquito-borne, emerging flavivirus. In addition to asymptomatic or mild febrile manifestations, ZIKV infection in humans is associated with comparatively rare congenital and neurological abnormalities. Previous in vitro studies have demonstrated cross-reactivity of antibodies derived from dengue virus infections with ZIKV, as well as antibody-mediated enhancement of infection in cell culture. Therefore, there is concern among vaccinologists and the public health community that the more severe disease manifestations could result from prior immunity, either from natural infection or vaccination, to antigenically-related flaviviruses (e.g., dengue, yellow fever, Japanese encephalitis). In this study, we evaluated the impact of prior immunity to dengue or yellow fever virus on ZIKV infection in a non-human primate model. After confirming that our immune cohorts were capable of recapitulating the published in vitro observations of cross-reactivity and enhancement, we looked for alteration of viral titers in peripheral blood, urine, cerebrospinal fluid, saliva, and vaginal secretions, as well as alterations in gross pathology, clinical parameters, and immune response, as compared to our flavivirus-naïve cohort. Importantly, no significant differences between flavivirus-naïve and primed animals were observed in viral titers or biofluid distribution, neutralizing antibody levels, or immune cell kinetics following ZIKV infection.