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Proteomics of human sputum from TB patients with high and low calprotectin

ABSTRACT: Sputum supernatant samples were collected from patients with and without active tuberculosis. Calprotectin was quantified and a total of twelve samples were selected for LC-MS/MS. Six samples were from patients with active TB and six samples were from TB-negative patients. Of the six samples from TB and non-TB sets, three samples had low concentrations of calprotectin and three samples had high concentrations of calprotectin. Overall, there were four groups with three samples each: non-TB with low CP, non-TB with high CP, TB with low CP, and TB with high CP. Each sample was prepared in technical triplicate, thus there were a total of 36 samples prepared for LC-MS/MS.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Sputum

DISEASE(S): Pulmonary Tuberculosis

SUBMITTER: Endrei Marcantonio

LAB HEAD: Sladjana Prisic

PROVIDER: PXD065067 | Pride | 2025-07-23

REPOSITORIES: Pride

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Project description:Novel biomarkers to identify infectious patients transmitting Mycobacterium tuberculosis are urgently needed to control the global tuberculosis (TB) pandemic. We hypothesized that proteins released into the plasma in active pulmonary TB are clinically useful biomarkers to distinguish TB cases from healthy individuals and patients with other respiratory infections. We applied a highly sensitive non-depletion tandem mass spectrometry discovery approach to investigate plasma protein expression in pulmonary TB cases compared to healthy controls in South African and Peruvian cohorts. . Plasma samples were analysed from 11 untreated male patients with active pulmonary TB and 10 male healthy control samples. Each plasma sample was initially separated into four segments by size exclusion chromatography, and each segment was processed individually. Analyses of plasma segments were performed in twelve iTRAQ (isobaric tags for relative and absolute quantification) 8-plex experimental sets in a block randomised design comprising three experimental sets. Each iTRAQ experiment contained a bridging master-pool plasma sample run in every experiment. Healthy controls were matched to TB samples by age, ethnicity, and smoking status within each iTRAQ set. Protein abundances from the plasma segments and multi-consensus reports were combined and adjusted for experimental batch effects. Protein abundances from the remaining combined plasma segment proteomes between experimental sets and the combined multi-consensus proteomes were analysed by complementary bioinformatic approaches to identify candidate diagnostic protein biomarkers. In total, 4,696 protein identifications were made across all iTRAQ experiments, at 5% FDR (false discovery rate). This comprised 2,332 unique host-derived proteins and 22 Mtb-derived proteins. Of these, 594 host proteins had a quantification result for every sample analysed and therefore comprised the complete quantified proteome. Subsequent bioinformatic analysis identified 118 differentially expressed proteins by three complementary bioinformatic pipelines and were taken forward in subsequent validation work as strong candidate biomarkers of pulmonary TB.

			Action	DRS
	NEG_HIGH_CP_Rep1_1.mzML	Mzml
	NEG_HIGH_CP_Rep1_2.mzML	Mzml
	NEG_HIGH_CP_Rep1_3.mzML	Mzml
	NEG_HIGH_CP_Rep2_1.mzML	Mzml
	NEG_HIGH_CP_Rep2_2.mzML	Mzml

Dataset Information

Proteomics of human sputum from TB patients with high and low calprotectin

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