Dataset Information

Mass spectrometry analysis of the blood plasma and serum samples from healthy donors

ABSTRACT: Previously, we published a dataset of human blood plasma and serum samples of 10 healthy males and 10 healthy females, fractionated on a set of sorbents (cation exchange Toyopearl CM-650M, CM Bio-Gel A, SP Sephadex C-25 and anion exchange QAE Sephadex A-25) and analyzed by LC-MS/MS individually and pooled in equal amounts (Supplementary Table S1, Sheet 1) [33]. We focused on those peptides that could be found in the blood plasma and serum using a standard search against the database of human proteins (UniProt Knowledgebase, taxon human). The combination of search engines MASCOT and X! Tandem, based on Scaffold 4 software with FDR less than 1%, allowed us to identify 15,530 unique peptides that belong to 2,127 protein groups (Table 1, Supplementary Table S1, Sheets 2 and 3). The Venn diagram shows that slightly less than 40% of the peptides are found in all four analyzed samples (Figures 1A and 1B). About 70% of the peptides are found in at least two samples. The accumulation diagram (Figure 1C) shows that the number of new unique peptides does not reach a plateau after repeated analyses of the same plasma sample — even after six runs. The high variability of the plasma and serum peptidome most likely indicates its sophisticated composition. 33. Arapidi, G. et al. Peptidomics dataset: Blood plasma and serum samples of healthy donors fractionated on a set of chromatography sorbents. Data Brief 18, 1204–1211 (2018).

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma, Blood Serum

DISEASE(S): Disease Free

SUBMITTER: Georgij Arapidi

LAB HEAD: Georgij Pavlovich Arapidi

PROVIDER: PXD027552 | Pride | 2024-06-14

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Description_of_samples.xlsx	Xlsx
	HealthyDonors_Serum-Plasma_UniProtKB-HS_72_2_Condensed.sf3	Other
	I305_20140905_SWP100_CMBiogel20_B1_5-40_120min_7uLinj_1.wiff	Wiff
	I305_20140905_SWP100_CMBiogel20_B1_5-40_120min_7uLinj_1.wiff.scan	Wiff
	I305_MGFPeaklist.dat	Other

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Project description:Human type 1 diabetes (T1D) arises through autoimmunity towards the insulin-producing pancreatic β cells and is modeled by the BioBreeding (BB) rat. Factors associated with islet autoimmunity are dilute and difficult to directly measure in the periphery. Therefore, we previously utilized microarray-based bioassay where human T1D sera were used to induce a disease-specific gene expression signature in unrelated, healthy peripheral blood mononuclear cells (PBMC). Here we report that sera of both spontaneously diabetic BB DRlyp/lyp and diabetes inducible BB DR+/+ rats induce inflammatory signatures in healthy PBMC of non-diabetic Brown Norway (BN) rats, while allogeneic BN sera does not. Consistent with their common T1D susceptibility, transcriptional signatures of both BB sub-strains share identity, including cytokines, immune receptors and signaling molecules. However, like the human T1D signature, the DRlyp/lyp signature, which is associated with progression to diabetes, is differentiated from that of the DR+/+ by induction of many IL-1 regulated genes. Addition of IL-1RA to cultures significantly ablates the DRlyp/lyp signature. Daily treatment of DRlyp/lyp rats with human recombinant IL-1RA significantly delays onset and sera of treated animals do not induce the characteristic IL-1 signature. Consistent with the presence of immune regulation in DR+/+ rats is induction of a signature showing negative regulation of the NFκB pathway. This study supports prior human investigations of serum that reflect disease processes associated with progression to T1D and add to a growing body of evidence implicating the balance of IL-1 and IL-1RA as key factors influencing the severity and outcome of inflammatory responses. Experiment Overall Design: The induction of gene expression was accomplished by culturing the PBMCs of healthy day180 BN donor rats for 6 h at 37°C in 5% CO2 with a) 20% autologous BN plasma(10 replicates), b) allogeneic BN plasma (15 replicates), c) allogeneic DRlyp/lyp plasma (day 60, 4 replicates), or d) DR+/+ plasma (day 60, 4 replicates). Healthy BN PBMCs were also cultured with d60 DRlyp/lyp serum supplemented with IL-1RA (4 replicates) as well as autologous BN serum supplemented with IL-1B (4 replicates), to respectively block or induce IL-1 mediated gene expression. To determine if IL-1RA-treatment of DRlyp/lyp rats would result in modulation of the DRlyp/lyp signature, additional DRlyp/lyp rats were treated with hIL-1RA (n=6) or PBS (n=6). Rats were treated for 10 days, beginning at d30, then serum was collected at d40 and used to induce gene expression in PBMCs isolated from healthy d180 BN rats. Thus, there are a total of 53 chips being the addition of all the sample conditions mentioned above.

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Mass spectrometry analysis of the blood plasma and serum samples from healthy donors

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