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ORIGINAL RESEARCH article

Front. Immunol.

Sec. Microbial Immunology

Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1636909

This article is part of the Research TopicHost-Pathogen Protein Interactions in Tuberculosis: Key Insights and ImplicationsView all articles

Neutrophil proteins as potential biomarkers for a sputum-based tuberculosis screening test

Provisionally accepted
  • 1Africa Health Research Institute (AHRI), Durban, South Africa
  • 2University of KwaZulu-Natal, Durban, South Africa
  • 3Reproductive Health and HIV Institute, Hillbrow, South Africa
  • 4Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, United States of America, Boston, United States
  • 5SAMRC Centre for Tuberculosis Research, Tygerberg, South Africa
  • 6Division of Infectious Diseases, Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee, USA, Nashville, United States
  • 7South African Tuberculosis Vaccine Initiative, Institute of Infectious Disease & Molecular Medicine and Division of Immunology, University of Cape Town, Cape Town, South Africa, Cape Town, South Africa
  • 8University College London, London, United Kingdom

The final, formatted version of the article will be published soon.

The development of a rapid and affordable assay to screen participants for additional testing could streamline TB screening in resource-limited settings and for community-wide health screens. Sputum remains the primary testing sample, making it potentially ideal for a screening testing. Neutrophils are highly expanded in sputum from individuals with pulmonary TB with high specificity and have potential as a biomarker for TB. Three neutrophil associated proteins, neutrophil gelatinase associated-lipocalin (NGAL), the protein heterodimer S100A8/A9 and the protein death ligand-1 (PDL-1), were measured in presumptive TB cases from participants attending a primary healthcare clinic in Durban, South Africa, using commercially available ELISAs on a total of 79 participants from a 109-participant cohort. Participants with microbiologically confirmed TB were sampled after 1 month of treatment. Proteins were also measured in tongue swab samples in participants from this cohort at baseline. Baseline results were confirmed in a second TB cohort which recruited a total of 51 participants with presumptive TB from the Western Cape. Finally, we investigate sputum neutrophil protein levels in individuals with community-diagnosed asymptomatic TB. Significant increases in all proteins were detectable in sputum from clinic-diagnosed TB participants relative to symptomatic controls. Performance approached the WHO target product profile for a TB triage test, with ROC AUCs reaching 0.866 (with a 95% confidence interval of 0.7683 -0.9633) in the case of S100A8/A9. Sputum protein levels did not correlate with bacterial burden and did not consistently decrease following one month of drug therapy. Only PDL-1 was detectable in mouth swab samples. Sputum neutrophil proteins tended to be elevated in participants with asymptomatic community diagnosed TB, as compared to asymptomatic community controls within the Vukuzazi cohort using a sample size of 42 participants, although this was not significant. This study provides a proof of principle that neutrophil proteins can be easily measured in standard sputum samples and have potential as a screening test for TB. However, more work is needed to explore whether this approach, using these three neutrophil proteins, can meet the WHO target product profile for a triage test worth developing further.

Keywords: Tuberculosis, Neutrophil proteins, Screening test, PDL1, ELISA -enzyme-linked immunosorbent assay. (Min

Received: 28 May 2025; Accepted: 06 Oct 2025.

Copyright: © 2025 Chambers, Karim, Mazibuko, Mhlane, Madziwa, Moosa, Moodley, Hoque, Wong, Hiemstra, Malherbe, Kriel, Stanley, van Rensburg, Shabangu, Smith, Walzl, Du Plessis, Sterling, Hatherill and Leslie. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Alasdair Leslie, al.leslie@ahri.org

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