Project description:Whole transcriptome RNA sequencing in brain tissue was generated to explore differences between young and old animals of two closely related species of deer mice (genus Peromyscus) that reportedly differ in their lifespans: P. leucopus that lives for up to 8 years and P. maniculatus that exhibits a lifespan of about 4 years.
Project description:Currently, most tools utilized in host-pathogen interaction studies depend on the use of human or mouse (Mus musculus) cells and tissues. While these species have led to countless breakthroughs in our understanding of infectious disease, there are undoubtably important biological processes that are missed by limiting studies to these two vertebrate species. For instance, it is well-established that the most common North American rodent, the Peromyscus leucopus deermouse, has unique interactions with microbes, which likely shape its ability to serve as a critical reservoir for at numerous zoonotic pathogens—including a Lyme disease spirochete, Borrelia burgdorferi. In this work, we expand the immunological toolkit to study P. leucopus biology by performing the first differentiation of deermouse bone marrow to macrophages using P. leucopus M-CSF producing HEK293T cells.
Project description:Epitope mapping studies aim to identify the binding sites of antibody-antigen interactions to enhance the development of vaccines, diagnostics and immunotherapeutic compounds. However, mapping is a laborious process employing time- and resource-consuming M-bM-^@M-^Xwet benchM-bM-^@M-^Y techniques or epitope prediction software that are still in their infancy. For polymorphic antigens, another challenge is characterizing cross-reactivity between epitopes, teasing out distinctions between broadly cross-reactive responses, limited cross-reactions among variants and the truly type-specific responses. A refined understanding of cross-reactive antibody binding could guide the selection of the most informative subsets of variants for diagnostics and multivalent subunit vaccines. We explored the antibody binding reactivity of sera from human patients and Peromyscus leucopus rodents infected with Borrelia burgdorferi to the polymorphic outer surface protein C (OspC), an attractive candidate antigen for vaccine and improved diagnostics for Lyme disease. We constructed a protein microarray displaying 23 natural variants of OspC and quantified the degree of cross-reactive antibody binding between all pairs of variants, using Pearson correlation calculated on the reactivity values using three independent transforms of the raw data: (1) logarithmic, (2) rank, and (3) binary indicators. We observed that the global amino acid sequence identity between OspC pairs was a poor predictor of cross-reactive antibody binding. Then we asked if specific regions of the protein would better explain the observed cross-reactive binding and performed in silico screening of the linear sequence and 3-dimensional structure of OspC. This analysis pointed to the C-terminal helix of the structure as a major determinant of type-specific cross-reactive antibody binding. We developed bioinformatics methods to systematically analyze the relationship between local sequence/structure variation and cross-reactive antibody binding patterns among variants of a polymorphic antigen, and this method can be applied to other polymorphic antigens for which immune response data is available for multiple variants. Antibody profiling was performed on sera from Borrelia burgdorferi infected and non-infected humans and Peromyscus leucopus rodents against 23 variants of the surface protein OspC . For infected human serum samples, the OspC type of the infecting B. burgdorferi strain is unknown; for experimentally-infected P. leucopus serum samples, it is known. Of human serum samples, 55 were from infected individuals and 25 from naive controls. Of P. leucopus serum samples, 23 were from infected individuals and 7 were from naive controls.
Project description:Novel coronavirus causing Covid-19 identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused pandemic in 2020. Although the virus and disease in humans has been thoroughly researched, so far there has not been animal model comparable to humans – genetically diverse species able to get infected and sick from Covid-19. The white-footed deermouse Peromyscus leucopus is a long-lived rodent and a key reservoir in North America for agents of several zoonoses including Lyme disease, babesiosis, anaplasmosis, and viral encephalitis. While persistently infected, this deermouse avoids apparent disability or diminished fitness. Its tolerance to infection with sometimes more than one pathogen makes P. leucopus comparable to bats. This study uses P. leucopus, LL colony stock, as a genetically diverse animal model for viral infection with SARS-CoV-2. We infected P. leucopus with SARS-CoV-2, collected plasma, lungs, and brain 3 and 6 days post-infection, and compared to control animals. P. leucopus mount an immune response against viral pathogens through production of neutralizing antibodies and genome-wide transcription of type I interferon stimulated genes in lungs compared to naïve animals. Viral RNA detection correlates with gene expression of type I interferon stimulated genes in response to viral infection in the brain. We report that diversity of outbred animals, their sex and age is reflected in the range of responses. These results show that P. leucopus is a viable animal model for SARS-CoV-2, particularly in research of viral infection of the brain.
Project description:Novel coronavirus causing Covid-19 identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused pandemic in 2020. Although the virus and disease in humans has been thoroughly researched, so far there has not been animal model comparable to humans – genetically diverse species able to get infected and sick from Covid-19. The white-footed deermouse Peromyscus leucopus is a long-lived rodent and a key reservoir in North America for agents of several zoonoses including Lyme disease, babesiosis, anaplasmosis, and viral encephalitis. While persistently infected, this deermouse avoids apparent disability or diminished fitness. Its tolerance to infection with sometimes more than one pathogen makes P. leucopus comparable to bats. This study uses P. leucopus, LL colony stock, as a genetically diverse animal model for viral infection with SARS-CoV-2. We infected P. leucopus with SARS-CoV-2, collected plasma, lungs, and brain 3 and 6 days post-infection, and compared to control animals. P. leucopus mount an immune response against viral pathogens through production of neutralizing antibodies and genome-wide transcription of type I interferon stimulated genes in lungs compared to naïve animals. Viral RNA detection correlates with gene expression of type I interferon stimulated genes in response to viral infection in the brain. We report that diversity of outbred animals, their sex and age is reflected in the range of responses. These results show that P. leucopus is a viable animal model for SARS-CoV-2, particularly in research of viral infection of the brain.
Project description:Novel coronavirus causing Covid-19 identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused pandemic in 2020. Although the virus and disease in humans has been thoroughly researched, so far there has not been animal model comparable to humans – genetically diverse species able to get infected and sick from Covid-19. The white-footed deermouse Peromyscus leucopus is a long-lived rodent and a key reservoir in North America for agents of several zoonoses including Lyme disease, babesiosis, anaplasmosis, and viral encephalitis. While persistently infected, this deermouse avoids apparent disability or diminished fitness. Its tolerance to infection with sometimes more than one pathogen makes P. leucopus comparable to bats. This study uses P. leucopus, LL colony stock, as a genetically diverse animal model for viral infection with SARS-CoV-2. We infected P. leucopus with SARS-CoV-2, collected plasma, lungs, and brain 3 and 6 days post-infection, and compared to control animals. P. leucopus mount an immune response against viral pathogens through production of neutralizing antibodies and genome-wide transcription of type I interferon stimulated genes in lungs compared to naïve animals. Viral RNA detection correlates with gene expression of type I interferon stimulated genes in response to viral infection in the brain. We report that diversity of outbred animals, their sex and age is reflected in the range of responses. These results show that P. leucopus is a viable animal model for SARS-CoV-2, particularly in research of viral infection of the brain.