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In vivo Host-Pathogen Interaction as Revealed by Global Proteomic Profiling of Zebrafish Larvae.

ABSTRACT: The outcome of a host-pathogen interaction is determined by the conditions of the host, the pathogen, and the environment. Although numerous proteomic studies of in vitro-grown microbial pathogens have been performed, in vivo proteomic approaches are still rare. In addition, increasing evidence supports that in vitro studies inadequately reflect in vivo conditions. Choosing the proper host is essential to detect the expression of proteins from the pathogen in vivo. Numerous studies have demonstrated the suitability of zebrafish (Danio rerio) embryos as a model to in vivo studies of Pseudomonas aeruginosa infection. In most zebrafish-pathogen studies, infection is achieved by microinjection of bacteria into the larvae. However, few reports using static immersion of bacterial pathogens have been published. In this study we infected 3 days post-fertilization (DPF) zebrafish larvae with P. aeruginosa PAO1 by immersion and injection and tracked the in vivo immune response by the zebrafish. Additionally, by using non-isotopic (Q-exactive) metaproteomics we simultaneously evaluated the proteomic response of the pathogen (P. aeruginosa PAO1) and the host (zebrafish). We found some zebrafish metabolic pathways, such as hypoxia response via HIF activation pathway, were exclusively enriched in the larvae exposed by static immersion. In contrast, we found that inflammation mediated by chemokine and cytokine signaling pathways was exclusively enriched in the larvae exposed by injection, while the integrin signaling pathway and angiogenesis were solely enriched in the larvae exposed by immersion. We also found important virulence factors from P. aeruginosa that were enriched only after exposure by injection, such as the Type-III secretion system and flagella-associated proteins. On the other hand, P. aeruginosa proteins involved in processes like biofilm formation, and cellular responses to antibiotic and starvation were enriched exclusively after exposure by immersion. We demonstrated the suitability of zebrafish embryos as a model for in vivo host-pathogen based proteomic studies in P. aeruginosa. Our global proteomic profiling identifies novel molecular signatures that give systematic insight into zebrafish-Pseudomonas interaction.

SUBMITTER: Diaz-Pascual F

PROVIDER: S-EPMC5524664 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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In vivo Host-Pathogen Interaction as Revealed by Global Proteomic Profiling of Zebrafish Larvae.

Díaz-Pascual Francisco F Ortíz-Severín Javiera J Varas Macarena A MA Allende Miguel L ML Chávez Francisco P FP

Frontiers in cellular and infection microbiology 20170725

The outcome of a host-pathogen interaction is determined by the conditions of the host, the pathogen, and the environment. Although numerous proteomic studies of in vitro-grown microbial pathogens have been performed, in vivo proteomic approaches are still rare. In addition, increasing evidence supports that in vitro studies inadequately reflect in vivo conditions. Choosing the proper host is essential to detect the expression of proteins from the pathogen in vivo< ...[more]

PMID: 28791256

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Project description:The interaction between fungal pathogen and host during infection is a complex and dynamic process. To resolve this, we chose the zebrafish model organism as the host to study C. albicans infection via systems biology approach. Transcriptome microarray data and histological analysis of surviving fish were sampled at different post-infection time points. The dynamic variations of significant genes expression profiles in C. albicans and zebrafish were concurrently analyzed by principal component analysis (PCA). The PCA results clearly indicated that the infection of C. albicans can be divided into three phases that include adhesion, invasion and damage phases. The results were highly consistent with subsequent histological analysis. Furthermore, we found the primary ontology function of genes with significant variations in both C. albicans and zebrafish is iron related. Most of the iron related genes in C. albicans were over-expressed in late stage, while most of the iron related genes in zebrafish were suppressed at the same phase of infection. It suggested that the iron homeostasis function of the host was shut-down when massive hemorrhage in zebrafish occurred during later stages of infection. At the same time, the iron scavenging function of C. albicans was activated. This implied the competition for iron is an important issue between the host and the fungal pathogen during infection. When we administered excess iron into the microenvironment of infection site, the infection process was significantly delayed. That indicated the virulence of C. albicans is correlated with its protein-bond iron scavenging strategies. Our finings not only provided dynamic mechanistic views of the iron competition but also highlighted the potential regulatory schemes in fungal pathogenesis. Each fish was intraperitoneally injected with C. albicans cells and these infected fish were collected at 0.5, 1, 2, 4, 6, 8, 12, 16, 18 hpi. 0.625M-NM-<g of Cy3 cRNA for C. albicans array and 1.65 M-NM-<g of Cy3 cRNA for zebrafish array was fragmented to an average size of about 50-100 nucleotides by incubation with fragmentation buffer at 60M-BM-0C for 30 minutes. Each time points contain three biological repeat. For C. albicans array, each biological repeat has two technical replicate.

Dataset Information

In vivo Host-Pathogen Interaction as Revealed by Global Proteomic Profiling of Zebrafish Larvae.

Publications

<i>In vivo</i> Host-Pathogen Interaction as Revealed by Global Proteomic Profiling of Zebrafish Larvae.

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