A meta-proteomics approach using autopsy material from the pre-antibiotic era from patients with untreated pulmonary tuberculosis to identify proteins present in early lesions of post-primary tuberculosis
Beatrice Normann, Syeda Mariam Riaz, Lisbet Sviland, Tehmina Mustafa, Even Birkeland
Abstract
Primary TB and post-primary TB (PPTB) are different disease entities. PPTB occurs only in humans, and no animal model mimics the actual pathology of PPTB. Information on the immune pathogenesis of PPTB is lacking due to the scarcity of untreated human material. In the early stages of PPTB, lesions can either progress or regress.
Introduction
Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis), with more than 80% of patients having pulmonary involvement. TB continues to remain a public health priority even after the discovery of effective and affordable antibiotics more than fifty years ago [1].
Materials and methods
Formalin-fixed paraffin-embedded (FFPE) blocks from 1931–47 were collected from the archives stored at the Department of Pathology, the Gades Institute, Haukeland University Hospital, Bergen, Norway, as a part of another study on 12-02-2019.
Results
There were 81 cases with lung paraffin blocks available. Only 5 cases with easily detectable lesions on polyethylene (PEN) membrane slides were selected for this pilot study. Table 1 shows the characteristics of each patient used in this study.
Discussion
Using the proteomics-based approach, we analyzed the proteins present in early and necrotic lesions of human PPTB from archival formalin-fixed and paraffin-embedded lung tissues collected between 1931–1947. In total, 3,531 human proteins and 110 bacterial proteins were identified, including five of mycobacterial origin, one from Mycobacterium avium and four from M. tuberculosis.
Conclusion
By using microdissection, TMT-labelling and mass spectrometry, we identified 3531 human,110 bacterial and fungal proteins from 80-year-old archival formalin-fixed and paraffin-embedded lung tissues of TB patients.
Citation: Normann B, Riaz SM, Sviland L, Mustafa T, Birkeland E (2026) A meta-proteomics approach using autopsy material from the pre-antibiotic era from patients with untreated pulmonary tuberculosis to identify proteins present in early lesions of post-primary tuberculosis. PLoS One 21(4): e0345052. https://doi.org/10.1371/journal.pone.0345052
Editor: Fumihiro Yamaguchi, Showa University Fujigaoka Hospital, JAPAN
Received: September 1, 2025; Accepted: February 27, 2026; Published: April 1, 2026
Copyright: © 2026 Normann et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: All relevant data are within the manuscript and its Supporting Information files.
Funding: Partial funding of the project was provided by the Blakstad Maarschalk og Helbings legat for bekjempelse av tuberkulose og kreft (UNIFOR). Partal funding was provided by the University of Bergen. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Abbreviations: TB, Tuberculosis; M. tuberculosis, Mycobacterium tuberculosis; FFPE, Formalin-fixed paraffin-embedded; MS/MS, Tandem mass spectrometry; m/z, Mass-to-charge ratio; H2O, Water; TCEP, Tris(2-carbocyethyl) phosphine; CAA, Chloroacetamide; BCA, Bicinchoninic acid; AmBic, Ammonium bicarbonate; CaCl2, Calcium chloride; HCL, Hydrochloric acid; NaCl, Sodium chloride; TFA, Trifluoracetic; ACN, acetonitrile; HE, Haematoxylin eosin; emPAI, Exponentially modified protein abundance index; PSM, Peptide-spectrum match
