ALBERT

Description: Approximately one fifth of all malignancies worldwide are etiologically associated with a persistent viral or bacterial infection. Thus, there is a particular interest in therapeutic molecules which use components of a natural immune response to specifically inhibit oncogenic microbial proteins, as it is anticipated they will elicit fewer off-target effects than conventional treatments. This concept has been explored in the context of human papillomavirus 16 (HPV16)-related cancers, through the development of monoclonal antibodies and fragments thereof against the viral E6 oncoprotein. Challenges related to the biology of E6 as well as the functional properties of the antibodies themselves appear to have precluded their clinical translation. Here, we addressed these issues by exploring the utility of the variable domains of camelid heavy-chain-only antibodies (denoted as VHHs). Through construction and panning of two llama, immune VHH phage display libraries, a pool of potential VHHs was isolated. The interactions of these with recombinant E6 were further characterized using an enzyme-linked immunosorbent assay (ELISA), Western blotting under denaturing and native conditions, and surface plasmon resonance. Three VHHs were identified that bound recombinant E6 with nanomolar affinities. Our results lead the way for subsequent studies into the ability of these novel molecules to inhibit HPV16-infected cells in vitro and in vivo.

Description: High-risk strains of human papillomavirus are causative agents for cervical and other mucosal cancers, with type 16 being the most frequent. Compared to the European Prototype (EP; A1), the Asian-American (AA; D2/D3) sub-lineage seems to have increased abilities to promote carcinogenesis. Here, we studied protein–protein interactions (PPIs) between host proteins and sub-lineages of the key transforming E6 protein. We transduced human keratinocyte with EP or AA E6 genes and co-immunoprecipitated E6 proteins along with interacting cellular proteins to detect virus–host binding partners. AAE6 and EPE6 may have unique PPIs with host cellular proteins, conferring gain or loss of function and resulting in varied abilities to promote carcinogenesis. Using liquid chromatography-mass spectrometry and stringent interactor selection criteria based on the number of peptides, we identified 25 candidates: 6 unique to AAE6 and EPE6, along with 13 E6 targets common to both. A novel approach based on pathway selection discovered 171 target proteins: 90 unique AAE6 and 61 unique EPE6 along with 20 common E6 targets. Interpretations were made using databases, such as UniProt, BioGRID, and Reactome. Detected E6 targets were differentially implicated in important hallmarks of cancer: deregulating Notch signaling, energetics and hypoxia, DNA replication and repair, and immune response.

Description: Facilitating voluntary conservation on private lands is a crucial element of policies that seek to mitigate forest habitat loss and fragmentation around the world. Previous research emphasizes the role of social factors (e.g., landowner characteristics, economics) in forest management, but environmental outcomes of past management can also affect landowner decisions. Our objective was to evaluate how positive outcomes for wildlife and habitat might reinforce or amplify landowner efforts to manage forest habitats. We applied the lens of coupled human and natural systems to investigate private lands management for early successional forests, which are declining along with associated wildlife in rural areas of the eastern U.S. Efforts to restore early successional forest in this region involve active forest management to create patches of successional forest in native, mature mixed hardwood stands. By integrating field-based monitoring of wildlife with surveys of landowner perceptions, we examined how landowners observed, interpreted, and responded to property-scale ecological outcomes of forest management. We recorded presence of Golden-winged Warbler (Vermivora chrysoptera) and American Woodcock (Scolopax minor) and estimated bird species richness in spring 2015 and/or 2016 on private properties located in the Appalachians (Maryland, New Jersey, Pennsylvania) and Upper Great Lakes (Minnesota, Wisconsin). These properties were enrolled in early successional forest management programs administered through the Natural Resources Conservation Service. Bird surveys were paired with landowner responses to a telephone survey conducted from January to May 2017 (n = 102). Most (71.6–81.6%) landowners’ perceptions of avian presence on their properties matched monitoring results. These perceptions were informed by personal observations and by outreach from agency partners and field technicians. Landowners who already completed their conservation program contracts (n = 85) continued managing early successional forests. Continued management for early successional habitat was positively associated with perceived benefits to birds, forest health, and scenery. Our findings give insight into how private landowners respond to environmental effects of forest management. We conclude that positive environmental outcomes of these conservation programs are related to continued early successional forest conservation by private landowners.