Project description:Lyme borreliosis (LB) is a common infection of domestic dogs in areas where there is enzootic transmission of the agent Borrelia burgdorferi. Immunodiagnostic assays for canine antibodies to B. burgdorferi are usually based on whole-cell preparations as substrate and, consequently, interpretation of results is confounded by antibody cross-reactivity between borrelial antigens and those of other bacterial species. To more broadly characterize the antibody responses to B. burgdorferi infection and to assess the diversity in those responses between individual dogs, we examined sera from 32 adult, colony-bred beagle dogs that were experimentally infected with B. burgdorferi through tick bites and compared those on a protein microarray with sera from uninfected dogs in their antibody reactivities to various recombinant chromosome- and plasmid-encoded B. burgdorferi proteins, including 24 of the serotype-defining OspC proteins of North America. The profiles of immunogenic proteins for the dogs were largely similar to those for humans and natural reservoir rodents. These included the decorin-binding protein DbpB, BBA36, BBA57, BBA64, fibronectin-binding protein BBK32, VlsE, FlaB and other flagellar structural proteins, Erp proteins, Bdr proteins, and all of the OspC proteins. In addition, the canine sera bound to the presumptive lipoproteins BBB14 and BB0844, which infrequently elicited antibodies in humans or rodents. Although the beagle, like most other domestic dog breeds, has a small effective population size and features extensive linkage disequilibrium, the group of animals here demonstrated diversity in antibody responses by measures of antibody levels and specificities for conserved proteins, like DbpB, and polymorphic ones, like OspC.
Project description:Lyme borreliosis (LB) is a common infection of domestic dogs in areas where there is enzootic transmission of the agent Borrelia burgdorferi. Immunodiagnostic assays for canine antibodies to B. burgdorferi are usually based on whole-cell preparations as substrate and, consequently, interpretation of results is confounded by antibody cross-reactivity between borrelial antigens and those of other bacterial species. To more broadly characterize the antibody responses to B. burgdorferi infection and to assess the diversity in those responses between individual dogs, we examined sera from 32 adult, colony-bred beagle dogs that were experimentally infected with B. burgdorferi through tick bites and compared those on a protein microarray with sera from uninfected dogs in their antibody reactivities to various recombinant chromosome- and plasmid-encoded B. burgdorferi proteins, including 24 of the serotype-defining OspC proteins of North America. The profiles of immunogenic proteins for the dogs were largely similar to those for humans and natural reservoir rodents. These included the decorin-binding protein DbpB, BBA36, BBA57, BBA64, fibronectin-binding protein BBK32, VlsE, FlaB and other flagellar structural proteins, Erp proteins, Bdr proteins, and all of the OspC proteins. In addition, the canine sera bound to the presumptive lipoproteins BBB14 and BB0844, which infrequently elicited antibodies in humans or rodents. Although the beagle, like most other domestic dog breeds, has a small effective population size and features extensive linkage disequilibrium, the group of animals here demonstrated diversity in antibody responses by measures of antibody levels and specificities for conserved proteins, like DbpB, and polymorphic ones, like OspC.
Project description:Lyme borreliosis (LB) is a tick-borne infection caused by Borrelia burgdorferi. Dogs are at high risk of exposure to ticks and tick-borne pathogens, including B. burgdorferi. Immunodiagnostic assays are usually based on whole-cell preparations of B. burgdorferi as substrate and, consequently, interpretation of results is confounded by antibody cross-reactivity between borrelial antigens and other bacterial species, as well as the anti-LB vaccination status of the dog. For this study, we examined sera from 33 dogs that were experimentally infected with B. burgdorferi through tick bite. These sera were compared with sera from uninfected dogs in their reactivities to 72 different recombinant B. burgdorferi antigens and 24 OspC protein types on a protein microarray. Amongst antigens frequently recognized by infected dogs were several known to be immunogens for humans, such as Decorin-binding protein A (BBA25), BBA64, fibronectin-binding protein (BBK32), VlsE, Erp and Bdr, CRASP proteins, OspC proteins and some flagellar antigens. Of special interest were the novel antigens BBB14 and BB0844, both hypothetical lipoproteins about which very little is currently known, and that were frequently and strongly recognized by infected dog sera. The antibody response of B. burgdorferi-infected dogs presents both similarities and differences from human counterparts, and both can be important for improvement of canine LB diagnosis and vaccine development. Antibody profiling was performed on sera from dogs experimentally-infected with B. burgdorferi and unexposed controls against antigens of B. burgdorferi. Thirty-three serum samples from experimental infections, and 5 unexposed controls were probed on a protein microarray displaying 24 OspC proteins of B. burgdorferi .
Project description:Lyme borreliosis (LB) is a tick-borne infection caused by Borrelia burgdorferi. Dogs are at high risk of exposure to ticks and tick-borne pathogens, including B. burgdorferi. Immunodiagnostic assays are usually based on whole-cell preparations of B. burgdorferi as substrate and, consequently, interpretation of results is confounded by antibody cross-reactivity between borrelial antigens and other bacterial species, as well as the anti-LB vaccination status of the dog. For this study, we examined sera from 33 dogs that were experimentally infected with B. burgdorferi through tick bite. These sera were compared with sera from uninfected dogs in their reactivities to 72 different recombinant B. burgdorferi polypeptides on a protein microarray. Amongst antigens frequently recognized by infected dogs were several known to be immunogens for humans, such as Decorin-binding protein A (BBA25), BBA64, fibronectin-binding protein (BBK32), VlsE, Erp and Bdr, CRASP proteins, OspC proteins and some flagellar antigens. Of special interest were the novel antigens BBB14 and BB0844, both hypothetical lipoproteins about which very little is currently known, and that were frequently and strongly recognized by infected dog sera. The antibody response of B. burgdorferi-infected dogs presents both similarities and differences from human counterparts, and both can be important for improvement of canine LB diagnosis and vaccine development. Antibody profiling was performed on sera from dogs experimentally-infected with B. burgdorferi and unexposed controls against antigens of B. burgdorferi. Thirty-three serum samples from experimental infections, and 6 unexposed controls were probed on a protein microarray displaying 72 unique proteins of B. burgdorferi .
Project description:Comparative oncology is a developing research discipline that is being used to assist our understanding of human neoplastic diseases. Companion canines are a preferred animal oncology model due to spontaneous tumor development and similarity to human disease at the pathophysiological level. We use a paired RNA sequencing (RNA-Seq)/microarray analysis of a set of four normal canine lymph nodes and ten canine lymphoma fine needle aspirates to identify technical biases and variation between the technologies and convergence on biological disease pathways. Surrogate Variable Analysis (SVA) provides a formal multivariate analysis of the combined RNA-Seq/microarray data set. Applying SVA to the data allows us to decompose variation into contributions associated with transcript abundance, differences between the technology, and latent variation within each technology. A substantial and highly statistically significant component of the variation reflects transcript abundance, and RNA-Seq proved more sensitive for detection of transcripts expressed at low levels. Latent random variation among RNA-Seq samples is also distinct in character from that impacting microarray samples. In particular, we observed variation between RNA-Seq samples that reflects transcript GC content. Platform-independent variable decomposition without a priori knowledge of the sources of variation using SVA represents a generalizable method for accomplishing cross-platform data analysis. We identified genes differentially expressed between normal lymph nodes of disease free dogs and a subset of the diseased dogs diagnosed with B-cell lymphoma using each technology. There is statistically significant overlap between the RNA-Seq and microarray sets of differentially expressed genes. Analysis of overlapping genes in the context of biological systems suggests elevated expression and activity of PI3K signaling in B-cell lymphoma biopsies compared with normal biopsies, consistent with literature describing successful use of drugs targeting this pathway in lymphomas. RNA was extracted from 10 lymphoma fine needle aspirates attained from companion canines. 4 normal lymph node samples were obtained from a Beagle breeding colony at Pfizer, including two samples that were taken from the same dog but different lymph nodes. This Series represents the Affymetrix gene expression only, not RNA-Seq referenced above. RNA-Seq data have been submitted to SRA as SRA059558.