8 results on '"Nolan, Lisa K."'
Search Results
2. Avian pathogenic Escherichia coli (APEC) infection alters bone marrow transcriptome in chickens.
- Author
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Hongyan Sun, Peng Liu, Nolan, Lisa K., and Lamont, Susan J.
- Subjects
CHICKEN diseases ,ESCHERICHIA coli ,BIRDS ,GENE expression ,CELL differentiation ,IMMUNE response - Abstract
Background: Avian pathogenic Escherichia coli (APEC) is a major cause of disease impacting animal health. The bone marrow is the reservoir of immature immune cells; however, it has not been examined to date for gene expression related to developmental changes (cell differentiation, maturation, programming) after APEC infection. Here, we study gene expression in the bone marrow between infected and non-infected animals, and between infected animals with mild (resistant) versus severe (susceptible) pathology, at two times post-infection. Results: We sequenced 24 bone marrow RNA libraries generated from the six different treatment groups with four replicates each, and obtained an average of 22 million single-end, 100-bp reads per library. Genes were detected as differentially expressed (DE) between APEC treatments (mild pathology, severe pathology, and mock-challenged) at a given time point, or DE between 1 and 5 days post-infection (dpi) within the same treatment group. Results demonstrate that many immune cells, genes and related pathways are key contributors to the different responses to APEC infection between susceptible and resistant birds and between susceptible and non-challenged birds, at both times post-infection. In susceptible birds, lymphocyte differentiation, proliferation, and maturation were greatly impaired, while the innate and adaptive immune responses, including dendritic cells, monocytes and killer cell activity, TLR- and NOD-like receptor signaling, as well as T helper cells and many cytokine activities, were markedly enhanced. The resistant birds' immune system, however, was similar to that of non-challenged birds. Conclusion: The DE genes in the immune cells and identified signaling models are representative of activation and resolution of infection in susceptible birds at both post-infection days. These novel results characterizing transcriptomic response to APEC infection reveal that there is combinatorial activity of multiple genes controlling myeloid cells, and B and T cell lymphopoiesis, as well as immune responses occurring in the bone marrow in these early stages of response to infection. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
3. Mutational and transcriptional analyses of an avian pathogenic Escherichia coli ColV plasmid.
- Author
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Skyberg, Jerod A., Johnson, Timothy J., and Nolan, Lisa K.
- Subjects
MICROBIAL mutation ,GENETIC transcription ,ESCHERICHIA coli ,PLASMIDS ,MICROBIAL virulence ,VIRUS diseases in poultry ,MICROBIAL genetics - Abstract
Background: Previously we described a 184-kb ColV plasmid, pAPEC-O2-ColV, that contributed to the ability of an E. coli to kill avian embryos, grow in human urine, and colonize the murine kidney. Here, the roles of several genes encoded by this plasmid in virulence were assessed using mutational and transcriptional analyses. Methods: Genes chosen for deletion were iss, tsh, iutA, iroN, sitA, and cvaB. In addition, a 35-kb region of the plasmid, containing iss, tsh, and the ColV and iro operons, along with a 15-kb region containing both the aerobactin and sit operons, were deleted. Mutants were compared to the wild-type (APEC O2) for lethality to chick embryos and growth in human urine. Expression of the targeted genes was also assessed under these same conditions using RT-PCR Results: No significant differences between the mutants and the wild-type in these phenotypic traits were detected. However, genes encoding known or predicted iron transport systems were up-regulated during growth in human urine, as compared to growth in LB broth, while iss, hlyF, and iroN were strongly up-regulated in chick embryos. Conclusion: While no difference was observed between the mutant strains and their wild-type parent in the phenotypic traits assayed, we reasoned that some compensatory virulence mechanism, insensitivity of the virulence assays, or other factor could have obscured changes in the virulence of the mutants. Indeed we found several of these genes to be up-regulated in human urine and/or in the chick embryo, suggesting that certain genes linked to ColV plasmids are involved in the establishment of avian extraintestinal infection. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
4. Avian pathogenic Escherichia coli (APEC) infection alters bone marrow transcriptome in chickens.
- Author
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Sun H, Liu P, Nolan LK, and Lamont SJ
- Subjects
- Animals, Chickens, Cluster Analysis, Gene Expression Profiling, Gene Expression Regulation, High-Throughput Nucleotide Sequencing, Organ Specificity genetics, Poultry Diseases metabolism, Reproducibility of Results, Signal Transduction, Bone Marrow Cells metabolism, Escherichia coli pathogenicity, Escherichia coli Infections veterinary, Poultry Diseases genetics, Poultry Diseases microbiology, Transcriptome
- Abstract
Background: Avian pathogenic Escherichia coli (APEC) is a major cause of disease impacting animal health. The bone marrow is the reservoir of immature immune cells; however, it has not been examined to date for gene expression related to developmental changes (cell differentiation, maturation, programming) after APEC infection. Here, we study gene expression in the bone marrow between infected and non-infected animals, and between infected animals with mild (resistant) versus severe (susceptible) pathology, at two times post-infection., Results: We sequenced 24 bone marrow RNA libraries generated from the six different treatment groups with four replicates each, and obtained an average of 22 million single-end, 100-bp reads per library. Genes were detected as differentially expressed (DE) between APEC treatments (mild pathology, severe pathology, and mock-challenged) at a given time point, or DE between 1 and 5 days post-infection (dpi) within the same treatment group. Results demonstrate that many immune cells, genes and related pathways are key contributors to the different responses to APEC infection between susceptible and resistant birds and between susceptible and non-challenged birds, at both times post-infection. In susceptible birds, lymphocyte differentiation, proliferation, and maturation were greatly impaired, while the innate and adaptive immune responses, including dendritic cells, monocytes and killer cell activity, TLR- and NOD-like receptor signaling, as well as T helper cells and many cytokine activities, were markedly enhanced. The resistant birds' immune system, however, was similar to that of non-challenged birds., Conclusion: The DE genes in the immune cells and identified signaling models are representative of activation and resolution of infection in susceptible birds at both post-infection days. These novel results characterizing transcriptomic response to APEC infection reveal that there is combinatorial activity of multiple genes controlling myeloid cells, and B and T cell lymphopoiesis, as well as immune responses occurring in the bone marrow in these early stages of response to infection.
- Published
- 2015
- Full Text
- View/download PDF
5. tkt1, located on a novel pathogenicity island, is prevalent in avian and human extraintestinal pathogenic Escherichia coli.
- Author
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Li G, Kariyawasam S, Tivendale KA, Wannemuehler Y, Ewers C, Wieler LH, Logue CM, and Nolan LK
- Subjects
- Animals, Bacterial Typing Techniques, Chickens microbiology, DNA, Bacterial genetics, Escherichia coli metabolism, Escherichia coli pathogenicity, Humans, Multilocus Sequence Typing, Peptides metabolism, Phylogeny, Plasmids, Sequence Analysis, DNA, Escherichia coli genetics, Escherichia coli Proteins genetics, Genomic Islands, Transketolase genetics
- Abstract
Background: Extraintestinal pathogenic Escherichia coli are important pathogens of human and animal hosts. Some human and avian extraintestinal pathogenic E. coli are indistinguishable on the basis of diseases caused, multilocus sequence and phylogenetic typing, carriage of large virulence plasmids and traits known to be associated with extraintestinal pathogenic E. coli virulence., Results: The gene tkt1 identified by a previous signature-tagged transposon mutagenesis study, was found on a 16-kb genomic island of avian pathogenic Escherichia coli (APEC) O1, the first pathogenic Escherichia coli strain whose genome has been completely sequenced. tkt1 was present in 39.6% (38/96) of pathogenic Escherichia coli strains, while only 6.25% (3/48) of E. coli from the feces of apparently healthy chickens was positive. Further, tkt1 was predominantly present in extraintestinal pathogenic E. coli belonging to the B2 phylogenetic group, as compared to extraintestinal pathogenic E. coli of other phylogenetic groups. The tkt1-containing genomic island is inserted between the metE and ysgA genes of the E. coli K12 genome. Among different extraintestinal pathogenic E. coli of the B2 phylogenetic group, 61.7% of pathogenic Escherichia coli, 80.6% of human uropathogenic E.coli and 94.1% of human neonatal meningitis-causing E. coli, respectively, harbor a complete copy of this island; whereas, only a few avian fecal E. coli strains contained the complete island. Functional analysis showed that Tkt1 confers very little transketolase activity but is involved in peptide nitrogen metabolism., Conclusion: These results suggest tkt1 and its corresponding genomic island are frequently associated with avian and human ExPEC and are involved in bipeptide metabolism.
- Published
- 2012
- Full Text
- View/download PDF
6. Spleen transcriptome response to infection with avian pathogenic Escherichia coli in broiler chickens.
- Author
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Sandford EE, Orr M, Balfanz E, Bowerman N, Li X, Zhou H, Johnson TJ, Kariyawasam S, Liu P, Nolan LK, and Lamont SJ
- Subjects
- Animals, Chickens microbiology, Escherichia coli Infections genetics, Host-Pathogen Interactions, Male, Oligonucleotide Array Sequence Analysis, Poultry Diseases microbiology, Spleen microbiology, Chickens genetics, Escherichia coli Infections veterinary, Poultry Diseases genetics, Spleen metabolism, Transcriptome
- Abstract
Background: Avian pathogenic Escherichia coli (APEC) is detrimental to poultry health and its zoonotic potential is a food safety concern. Regulation of antimicrobials in food-production animals has put greater focus on enhancing host resistance to bacterial infections through genetics. To better define effective mechanism of host resistance, global gene expression in the spleen of chickens, harvested at two times post-infection (PI) with APEC, was measured using microarray technology, in a design that will enable investigation of effects of vaccination, challenge, and pathology level., Results: There were 1,101 genes significantly differentially expressed between severely infected and non-infected groups on day 1 PI and 1,723 on day 5 PI. Very little difference was seen between mildly infected and non-infected groups on either time point. Between birds exhibiting mild and severe pathology, there were 2 significantly differentially expressed genes on day 1 PI and 799 on day 5 PI. Groups with greater pathology had more genes with increased expression than decreased expression levels. Several predominate immune pathways, Toll-like receptor, Jak-STAT, and cytokine signaling, were represented between challenged and non-challenged groups. Vaccination had, surprisingly, no detectible effect on gene expression, although it significantly protected the birds from observable gross lesions. Functional characterization of significantly expressed genes revealed unique gene ontology classifications during each time point, with many unique to a particular treatment or class contrast., Conclusions: More severe pathology caused by APEC infection was associated with a high level of gene expression differences and increase in gene expression levels. Many of the significantly differentially expressed genes were unique to a particular treatment, pathology level or time point. The present study not only investigates the transcriptomic regulations of APEC infection, but also the degree of pathology associated with that infection. This study will allow for greater discovery into host mechanisms for disease resistance, providing targets for marker assisted selection and advanced drug development.
- Published
- 2011
- Full Text
- View/download PDF
7. Common and specific genomic sequences of avian and human extraintestinal pathogenic Escherichia coli as determined by genomic subtractive hybridization.
- Author
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Kariyawasam S, Scaccianoce JA, and Nolan LK
- Subjects
- Animals, Chickens microbiology, DNA, Bacterial chemistry, Escherichia coli classification, Escherichia coli isolation & purification, Humans, Hybridization, Genetic, Phylogeny, Sequence Analysis, DNA, Species Specificity, Turkeys microbiology, Escherichia coli genetics, Genome, Bacterial
- Abstract
Background: Suppression subtractive hybridization (SSH) strategy was used with extraintestinal pathogenic Escherichia coli (EXPEC) that cause avian colibacillosis (avian pathogenic E. coli or APEC) and human urinary tract infections (uropathogenic E. coli or UPEC) to determine if they possessed genes that were host and/or niche specific. Both APEC and UPEC isolates were used as tester and driver strains in 4 different SSHs in order to obtain APEC- and UPEC-specific subtraction fragments (SFs)., Results: These procedures yielded a total of 136 tester-specific SFs of which 85 were APEC-derived and 51 were UPEC-derived. Most of the APEC-derived SFs were associated with plasmids; whereas, the majority of UPEC-derived sequences matched to the bacterial chromosome. We further determined the distribution of these tester-derived sequences in a collection of UPEC and APEC isolates using polymerase chain reaction techniques. Plasmid-borne, APEC-derived sequences (tsh, cvaB, traR, traC and sopB) were predominantly present in APEC, as compared to UPEC. Of the UPEC-derived SFs, those encoding hemolysin D and F1C major and minor fimbrial subunits were present only in UPEC. However, two UPEC-derived SFs that showed strong similarity to the uropathgenic-specific protein gene (usp) occurred in APEC, demonstrating that usp is not specific to UPEC., Conclusion: This study provides evidence of the genetic variability of ExPEC as well as genomic similarities between UPEC and APEC; it did not identify any single marker that would dictate host and/or niche specificity in APEC or UPEC. However, further studies on the genes that encode putative or hypothetical proteins might offer important insight into the pathogenesis of disease, as caused by these two ExPEC.
- Published
- 2007
- Full Text
- View/download PDF
8. Characterizing the APEC pathotype.
- Author
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Rodriguez-Siek KE, Giddings CW, Doetkott C, Johnson TJ, and Nolan LK
- Subjects
- Animals, Bacterial Proteins classification, Bacterial Proteins genetics, Birds microbiology, Escherichia coli classification, Feces microbiology, Genes, Bacterial, Genotype, Lactose metabolism, Serotyping, Virulence genetics, Bird Diseases microbiology, Escherichia coli genetics, Escherichia coli pathogenicity
- Abstract
The purpose of this study was to compare avian pathogenic Escherichia coli (APEC) isolates to fecal isolates of apparently healthy poultry (avian fecal E. coli or AFEC) by their possession of various traits in order to ascertain whether APEC and AFEC are distinct and if the APEC strains constitute a distinct pathotype. Four hundred and fifty-one APEC and one hundred and four AFEC isolates were examined for possession of traits associated with the virulence of human extraintestinal pathogenic E. coli (ExPEC) as well as APEC. Several of the genes occurred in the majority of APEC and only infrequently in AFEC, including cvaC, iroN, iss, iutA, sitA, tsh, fyuA, irp2, and ompT. Of these genes, several have been found on large plasmids in APEC. Other genes occurred in significantly more APEC than AFEC but did not occur in the majority of APEC. Isolates were also evaluated by serogroup, lactose utilization, and hemolytic reaction. Twenty-nine and a half percent of the APEC and forty-two and three tenths percent of the AFEC were not serogrouped because they were not typeable with standard antisera, typed to multiple serogroups, were rough, autoagglutinated, or were not done. Around 65% of the typeable APEC (205 isolates) and AFEC (41 isolates) were classified into shared serogroups, and about a third of both fell into APEC- (113 isolates) or AFEC- (19 isolates) unique serogroups. Most were able to use lactose. No isolate was hemolytic. Overall, the majority of the APEC isolates surveyed shared a common set of putative virulence genes, many of which have been localized to an APEC plasmid known as pTJ100. This common set of genes may prove useful in defining an APEC pathotype.
- Published
- 2005
- Full Text
- View/download PDF
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