Spatial distribution of Mycobacterium tuberculosis mRNA and secreted antigens in acid-fast negative human antemortem and resected tissueResearch in context

Kievershen Nargan; Joel N. Glasgow; Sajid Nadeem; Threnesan Naidoo; Gordon Wells; Robert L. Hunter; Anneka Hutton; Kapongo Lumamba; Mpumelelo Msimang; Paul V. Benson; Adrie J.C. Steyn

EBioMedicine (Jul 2024)

Spatial distribution of Mycobacterium tuberculosis mRNA and secreted antigens in acid-fast negative human antemortem and resected tissueResearch in context

Kievershen Nargan,
Joel N. Glasgow,
Sajid Nadeem,
Threnesan Naidoo,
Gordon Wells,
Robert L. Hunter,
Anneka Hutton,
Kapongo Lumamba,
Mpumelelo Msimang,
Paul V. Benson,
Adrie J.C. Steyn

Affiliations

Kievershen Nargan: Africa Health Research Institute, University of KwaZulu-Natal, Durban, South Africa
Joel N. Glasgow: Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, USA
Sajid Nadeem: Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, USA
Threnesan Naidoo: Africa Health Research Institute, University of KwaZulu-Natal, Durban, South Africa; Department of Forensic and Legal Medicine, Walter Sisulu University, Mthatha, South Africa
Gordon Wells: Africa Health Research Institute, University of KwaZulu-Natal, Durban, South Africa
Robert L. Hunter: Department of Pathology and Laboratory Medicine, University of Texas Health Sciences Center at Houston, Houston, TX, USA
Anneka Hutton: Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA
Kapongo Lumamba: Africa Health Research Institute, University of KwaZulu-Natal, Durban, South Africa
Mpumelelo Msimang: Department of Anatomical Pathology, National Health Laboratory Service, IALCH, Durban, South Africa
Paul V. Benson: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA
Adrie J.C. Steyn: Africa Health Research Institute, University of KwaZulu-Natal, Durban, South Africa; Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, USA; Centers for AIDS Research and Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL, USA; Corresponding author. University of Alabama at Birmingham, Birmingham, AL, USA; Africa Health Research Institute, University of KwaZulu-Natal, Durban, South Africa.

Journal volume & issue: Vol. 105
p. 105196

Abstract

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Summary: Background: The ability to detect evidence of Mycobacterium tuberculosis (Mtb) infection within human tissues is critical to the study of Mtb physiology, tropism, and spatial distribution within TB lesions. The capacity of the widely-used Ziehl-Neelsen (ZN) staining method for identifying Mtb acid-fast bacilli (AFB) in tissue is highly variable, which can limit detection of Mtb bacilli for research and diagnostic purposes. Here, we sought to circumvent these limitations via detection of Mtb mRNA and secreted antigens in human tuberculous tissue. Methods: We adapted RNAscope, an RNA in situ hybridisation (RISH) technique, to detect Mtb mRNA in ante- and postmortem human TB tissues and developed a dual ZN/immunohistochemistry staining approach to identify AFB and bacilli producing antigen 85B (Ag85B). Findings: We identified Mtb mRNA within intact and disintegrating bacilli as well as extrabacillary mRNA. Mtb mRNA was distributed zonally within necrotic and non-necrotic granulomas. We also found Mtb mRNA within, and adjacent to, necrotic granulomas in ZN-negative lung tissue and in Ag85B-positive bronchiolar epithelium. Intriguingly, we observed accumulation of Mtb mRNA and Ag85B in the cytoplasm of host cells. Notably, many AFB were negative for Ag85B staining. Mtb mRNA was observed in ZN-negative antemortem lymph node biopsies. Interpretation: RNAscope and dual ZN/immunohistochemistry staining are well-suited for identifying subsets of intact Mtb and/or bacillary remnants in human tissue. RNAscope can identify Mtb mRNA in ZN-negative tissues from patients with TB and may have diagnostic potential in complex TB cases. Funding: Wellcome Leap Delta Tissue Program, Wellcome Strategic Core Award, the National Institutes of Health (NIH, USA), the Mary Heersink Institute for Global Health at UAB, the UAB Heersink School of Medicine.

Published in EBioMedicine

ISSN: 2352-3964 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Medicine: Medicine (General)
Website: http://www.journals.elsevier.com/ebiomedicine/

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