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Distinct transcriptional profiles of Leptospira borgpetersenii serovar Hardjo strains JB197 and HB203 cultured at different temperatures

ABSTRACT: Leptospirosis is a global zoonotic, neglected tropical disease. Interestingly, a high level of species specificity (both bacteria and host) plays a major role in the severity of disease presentation which can vary from asymptomatic to multi-organ failure. Pathogenic Leptospira colonize the kidneys of infected individuals and are shed in urine into the environment where they can survive until they are contracted by another host. This study looks at two strains of L. borgpetersenii, HB203 and JB197 which are genetically very similar, and identical by serotyping as serovar Hardjo, yet HB203 causes a chronic infection in the hamster while JB197 causes organ failure and mortality. To better characterize bacterial factors causing different disease outcomes, we examined the gene expression profile of these strains in the context of temperatures that would reflect natural Leptospira life cycles (environmentally similar 29oC and 37oC which is more indicative of host environment). We found vast differences in gene expression both between the strains and within strains between temperatures. Characterization of the transcriptome of L. borgpetersenii serovar Hardjo strains JB197 and HB203 provides insights into factors that can determine acute versus chronic disease in the hamster model of infection. Additionally, these studies highlight strain to strain variability within the same species, and serovar, at different growth temperatures, which needs to be considered when serovars are selected and propagated for use as bacterin vaccines used to immunize domestic animal species.

ORGANISM(S): Leptospira borgpetersenii serovar Hardjo

PROVIDER: GSE168507 | GEO | 2021/04/28

REPOSITORIES: GEO

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Proteome comparison of Leptospira borgpetersenii serovar Hardjo strains cultured at different temperatures

Project description:Leptospirosis, caused by bacteria of the genus Leptospira, is a zoonotic disease affecting humans, companion animals, and all major livestock species. Typical propagation of the highly fastidious Leptospira borgepetesenii serovar Hardjo is limited to 29°C. However, newer culture media formulations now facilitate isolation and propagation at 37°C, a temperature that more closely emulates in vivo conditions and is hypothesized to regulate the expression of virulence factors during host infection. Since protein expression by leptospires is temperature dependent, and therefore the proteome of bacterin vaccines can differ whether grown at 37°C compared to 29°C, we compared the proteome of strains of Leptospira borgpetersenii serovar Hardjo at each temperature; two well-established strains that causes acute (strain JB197) or chronic asymptomatic disease (strain HB203) in the hamster challenge model of leptospirosis and two more recently isolated strains designated TC129 and TC273 (both of which cause chronic asymptomatic disease in the hamster). We found proteomic expression differences within strains propagated at the routine temperature of 29°C, and compared to the newly achieved culture temperature of 37°C. Results highlight significant differential protein expression, including virulence factors, amongst identical serovars of L. borgpetersenii when propagated at 29oC, the collective variation of which can be diminished when propagated at 37oC. Collectively, there is increasingly more evidence available to suggest bacterin vaccine design would benefit from consideration of strains employed, and potential effects of growth temperature related to specific behavior of pathogens in vaccine composition.

2022-05-20 | PXD032831 | Pride

Analysis of the human endothelial cell response to pathogenic Leptospira interrogans vs. non-pathogenic Leptospira biflexa (HMEC data)

2010-07-27 | GSE23173 | GEO

Analysis of the human endothelial cell response to pathogenic Leptospira interrogans vs. non-pathogenic Leptospira biflexa (Eahy1 data)

2010-07-27 | GSE23172 | GEO

Adaptation of pathogenic Leptospira to oxidative stress

Project description:Pathogenic Leptospira spp. are the causative agents of the zoonotic disease leptospirosis. During infection, Leptospira are confronted with deadly reactive oxygen species (ROS). Withstanding ROS produced by the host innate immunity is an important strategy evolved by pathogenic Leptospira for persisting in and colonizing hosts. The peroxide stress regulator, PerR, represses genes involved in ROS defenses in L. interrogans. We have performed RNA sequencing in WT and perR mutant strains to characterize the L. interrogans adaptive response to hydrogen peroxide. We showed that Leptospira solicit three main peroxidase machineries (catalase, cytochrome C peroxidase and peroxiredoxin) and heme to adapt to peroxide stress as well as canonical chaperones of the heat shock response, and DNA repair. Determining the PerR regulon allowed to identify the PerR-dependent mechanisms of the peroxide adaptive response and has revealed a regulatory network involving other transcriptional regulators, two-component systems and sigma factors as well as non-coding RNAs that putatively orchestrate, in concert with PerR, this adaptive response. Our findings provide comprehensive insight into the mechanisms required by pathogenic Leptospira to overcome infection-related oxidants. This will participate in framing future hypothesis-driven studies to identify and decipher novel virulence mechanisms.

2020-09-22 | GSE140019 | GEO

Interplay between two peroxide stress regulators in the oxidative stress response and virulence of pathogenic Leptospira

Project description:Pathogenic Leptospira spp. are the causative agents of the zoonotic disease leptospirosis. During infection, Leptospira are confronted with deadly reactive oxygen species (ROS). Withstanding ROS produced by the host innate immunity is an important strategy evolved by pathogenic Leptospira for persisting in and colonizing hosts. The peroxide stress regulator, PerRA, represses genes involved in ROS defenses in L. interrogans. We have identified an ORF encoding a putative second PerR in pathogenic Leptospira that we named PerRB. We have determined the transcriptomic profil of a single perRB and a double perRAperRB mutants. The concomitant inactivation of perRA and perRB has a pleiotropic effect on the transcriptomic profil of L. interrogans. The lack of both PerRA and PerRB regulators led to the differential expression of several virulence-associated genes and a loss of virulence. Our findings provide new insights into a new regulatory network that controls virulence and host colonization.

2021-10-07 | GSE158051 | GEO

Transcriptional response of Leptospira interrogans to iron limitation and characterization of putative Fur-like protein

2010-03-22 | GSE20422 | GEO

Transcriptional response of Leptospira interrogans to iron limitation and characterization of putative Fur-like protein

Project description:Leptospirosis is a globally significant zoonotic disease caused by Leptospira spp. Iron is essential for growth of most bacterial species. Since availability of iron is low in the host, pathogens have evolved complex iron acquisition mechanisms to survive and establish infection. Virulence genes in some bacteria have been shown to be iron-regulated. In many bacteria, expression of iron-uptake and storage proteins is regulated by Fur. L. interrogans encodes four predicted Fur homologs; we have constructed a mutant in one of these, la1857. We conducted microarray analysis to identify differentially expressed genes in L. interrogans serovar Manilae grown under low iron compared with normal iron conditions found in EMJH medium. We also compared the transcriptional profile of the la1857 mutant versus wild-type Manilae under normal and low iron conditions. Analysis used RNA derived from L. interrogans serovar Manilae grown under low iron (EMJH medium + 40 ÂµM 2,2-dipyridyl) and normal iron as experimental and control samples, respectively. We have a serovar Manilae mutant in la1857, one of four predicted fur homologs. The mutant was also grown under low iron and normal iron conditions and compared with wild-type Manilae grown under low and normal iron conditions respectively. Three independent RNA samples (biological replicates) from parent and mutant strains grown with or without 2,2-dipyridyl were compared in a loop design resulting in 12 arrays. The orientation of Cy3 to Cy5 labeling was the same for replicates 1 and 3, while replicate 2 was a dye swap. Direct comparisons were made between arrays of experimental and control samples using raw data pulled from two different channels for data analysis. Data from wild-type Manilae grown under low iron versus normal iron and mutant versus wild-type under normal iron conditions is reported in the manuscript.

2010-05-05 | E-GEOD-20422 | biostudies-arrayexpress

A novel hybrid two-component system Lvr mediates virulence and global regulation in pathogenic Leptospira

Project description:Transmission of leptospirosis requires that the spirochete pathogen adapts rapidly to the mammalian host milieu during infection and the external environment upon shedding from the renal tubules of animal reservoirs. The pathogenic Leptospira genomes encode a notably large number (≥76) of putative two-component system (TCS) proteins, which presumably play a key role in switching the ecological niches. Yet to date, the regulatory networks that govern virulence and environmental adaptation have not been elucidated in Leptospira. We identified seven ORFs encoding putative TCS proteins that are exclusively conserved in all the pathogenic Leptospira species. Two of these ORFs (LMANv2_670020, LMANv2_670019), juxtaposed in an operon, are predicted to encode a cytoplasmic hybrid histidine kinase and response regulator. Corresponding transposon mutants ΔlvrA/B and ΔlvrB demonstrated a loss-of-virulence in a hamster model of leptospirosis and aberrant motility phenotype. RNA sequencing revealed the differential expression of transcripts on a global scale in ΔlvrA/B (8.13%), ΔlvrB (5.06%) and ΔlvrA/B∩ΔlvrB (8.24%). In vitro transcriptomes provided insights into the role of Lvr dyad in regulating virulence, motility, signal transduction and metabolism related genes. Phosphotransfer assays indicated that LvrA interacts with LvrB in a branched signaling pathway. Phylogenetic analyses indicated that Our findings suggest that a novel, hybrid two-component system Lvr plays a key role in governing virulence and mediating global regulation in pathogenic Leptospira.

2018-04-04 | GSE79107 | GEO

Analysis of the human endothelial cell response to pathogenic Leptospira interrogans vs. non-pathogenic Leptospira biflexa (HMEC data)

Project description:The overall goal of these experiments was to determine how human endothelial cells respond to pathogenic Leptospira interrogans. Leptospira interrogans causes leptospirosis, the most widespread zoonotic infection in the world. A hallmark of leptospirosis is widespread endothelial damage, which in severe cases leads to hemorrhage. In these experiments, we infected two endothelial cell lines with pathogenic Leptospira interrogans serovar Canicola strain Ca12-005, and as controls, with the non-pathogenic Leptospira biflexa serovar Patoc strain Pfra. As additional controls, uninfected cells were also included in the analyses. The cell line used fhere was a microvascular endothelial line, HMEC (Ades et al, 1992. HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol. 99:683-690); due to loss of the original analysis files, only raw data files are provided. Infection times were performed at a multiplicity of infection (# bacteria/endothelial cell) of 10 for either 1 hour or 3 hours, after which RNA was harvested and reverse transcribed. Labeled cDNAs were used to probe HEEBO arrays purchased from Microarrays Inc. (Nashville, TN). In each of three biological replicate experiments, for each time point, three comparisons were made. First, the L. interrogans-infected cells were compared to the L. biflexa-infected cells. Second, the L. Interrogans-infected cells were compared to the uninfected cells. Third, the L. biflexa-infected cells were compared to the uninfected cells. A second endothelial cell line,

2010-07-27 | E-GEOD-23173 | biostudies-arrayexpress

Analysis of the human endothelial cell response to pathogenic Leptospira interrogans vs. non-pathogenic Leptospira biflexa (Eahy1 data)

Project description:The overall goal of these experiments was to determine how human endothelial cells respond to pathogenic Leptospira interrogans. Leptospira interrogans causes leptospirosis, the most widespread zoonotic infection in the world. A hallmark of leptospirosis is widespread endothelial damage, which in severe cases leads to hemorrhage. In these experiments, we infected two endothelial cell lines with pathogenic Leptospira interrogans serovar Canicola strain Ca12-005, and as controls, with the non-pathogenic Leptospira biflexa serovar Patoc strain Pfra. As additional controls, uninfected cells were also included in the analyses. The cell line used was Ea.hy926, a macrovascular line (Edgell, C. J.,et al. 1990. In vitro Cell. & Dev. Biol. 26:1167-1172, and Edgell, C. J., et al. 1983. Proc. Natl. Acad. Sci. 80:3734-3737). Infection times were performed at a multiplicity of infection (# bacteria/endothelial cell) of 10 for either 1 hour or 3 hours, after which RNA was harvested and reverse transcribed. Labeled cDNAs were used to probe HEEBO arrays purchased from Microarrays Inc. (Nashville, TN). In each of three biological replicate experiments, for each time point, three comparisons were made. First, the L. interrogans-infected cells were compared to the L. biflexa-infected cells. Second, the L. Interrogans-infected cells were compared to the uninfected cells. Third, the L. biflexa-infected cells were compared to the uninfected cells. A second endothelial cell line, HMEC (Ades et al, 1992. HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol. 99:683-690), which is of microvascular origin, was also used; raw data files are provided separately.

2010-07-27 | E-GEOD-23172 | biostudies-arrayexpress

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