Dataset Information

Multi-omics analysis of Histoplasma capsulatum treated with monoclonal antibodies

ABSTRACT: Multi-omics analysis of Histoplasma capsulatum treated with monoclonal antibodies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Histoplasma Capsulatum G217b

TISSUE(S): Yeast

DISEASE(S): Histoplasmosis

SUBMITTER: Ernesto Nakayasu

LAB HEAD: Ernesto Nakayasu

PROVIDER: PXD017864 | Pride | 2021-06-16

REPOSITORIES: Pride

ACCESS DATA

Publications

Remodeling of the <i>Histoplasma Capsulatum</i> Membrane Induced by Monoclonal Antibodies.

Burnet Meagan C MC Zamith-Miranda Daniel D Heyman Heino M HM Weitz Karl K KK Bredeweg Erin L EL Nosanchuk Joshua D JD Nakayasu Ernesto S ES

Vaccines 20200602 2

Antibodies play a central role in host immunity by directly inactivating or recognizing an invading pathogen to enhance different immune responses to combat the invader. However, the cellular responses of pathogens to the presence of antibodies are not well-characterized. Here, we used different mass spectrometry techniques to study the cellular responses of the pathogenic fungus <i>Histoplasma capsulatum</i> to monoclonal antibodies (mAb) against HSP60, the surface protein involved in infection ...[more]

PMID: 32498228

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Project description:Histoplasma capsulatum is a thermally dimorphic fungus with worldwide distribution, and high incidence in the Americas. It is the etiologic agent of histoplasmosis, an important life-threatening systemic mycosis. Dimorphism is an important feature for fungal survival in different environments and it has been related to the virulence of H. capsulatum, and essential to the establishment of infection. Proteomic profiles have brought important contributions to the knowledge of metabolism and pathogenicity in several biological models. However, studies of the H. capsulatum proteome have been underexplored. In the present study, we report the first proteomic comparison between the mycelium and the yeast cells of H. capsulatum. Liquid chromatography coupled to mass spectrometry was used to evaluate the proteomic profile of the two phases of H. capsulatum. In summary, 214 proteins were only detected/or preferentially abundant in mycelium, while the same occurred to 335 proteins in yeast cells. In mycelium, enzymes related to the glycolytic pathway and to the alcoholic fermentation showed greater abundance, suggesting a higher use of anaerobic pathways for energy production. In yeast cells, proteins related to the tricarboxylic acid cycle and response to temperature stress showed high abundance. Proteins related to oxidative stress response or involved with cell wall metabolism were identified with differential abundance in both conditions. Validation of proteomic data was performed by enzymatic activity determination, western blot assays, or immunofluorescence microscopy. These experiments corroborated, directly or indirectly, the abundance of isocitrate lyase, 2-methylcitrate synthase, catalase B, and mannosyl-oligosaccharide-1,2-alpha-mannosidase in the mycelium and heat shock protein (HSP) 30, HSP60, glucosamine-fructose-6-phosphate aminotransferase, glucosamine-6-phosphate deaminase, and N-acetylglucosamine-phosphate mutase in yeast-cells. The proteomic profile associated functional classification analyzes of proteins provided a better understanding of the metabolic reorganization and cell wall remodeling on the yeast form of H. capsulatum.

Project description:The fungal pathogen Histoplasma capsulatum is thought to be the most common cause of fungal respiratory infections in immunocompetent humans, yet little is known about its biology. Here we provide the first genome-wide studies to experimentally validate its genome annotation. A functional interrogation of the Histoplasma genome provides critical support for continued investigation into the biology and pathogenesis of H. capsulatum and related fungi. We employed a three-pronged approach to provide a functional annotation for the H. capsulatum G217B strain. First, we probed high-density tiling arrays with labeled cDNAs from cells grown under diverse conditions. These data defined 6,172 transcriptionally active regions (TARs), providing validation of 6,008 gene predictions. Interestingly, 22% of these predictions showed evidence of anti-sense transcription. Additionally, we detected transcription of 264 novel genes not present in the original gene predictions. To further enrich our analysis, we incorporated expression data from whole-genome oligonucleotide microarrays. These expression data included profiling under growth conditions that were not represented in the tiling experiment, and validated an additional 2,249 gene predictions. Finally, we compared the G217B gene predictions to other available fungal genomes, and observed that an additional 254 gene predictions had an ortholog in a different fungal species, suggesting that they represent genuine coding sequences. These analyses yielded a high confidence set of validated gene predictions for H. capsulatum. The transcript sets resulting from this study are a valuable resource for further experimental characterization of this ubiquitous fungal pathogen. The data is available for interactive exploration at http://histo.ucsf.edu. The non-redundant genome sequence of Histoplasma capsulatum G217B was tiled over a set of 93 CombiMatrix arrays, which were then hybridized with labeled cDNA samples from yeast-form (red channel) or mycelial-form (green channel) Histoplasma. Due to low yields from the mycelial samples, only the red channel intensities were analyzed (and the red foreground intensities are, therefore, reported in the VALUE column for each sample). Rather than normalizing intensities across arrays, each probe was evaluated as detected or undetected relative to the negative control intensities on the corresponding array, and the density of detected probes as a function of genome position was used for the remaining analysis.

Dataset Information

Multi-omics analysis of Histoplasma capsulatum treated with monoclonal antibodies

Publications

Remodeling of the <i>Histoplasma Capsulatum</i> Membrane Induced by Monoclonal Antibodies.

Similar Datasets

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