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2. Division of Microbiology and Molecular Medicine, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, Linköping, Sweden.
3. Phagocyte Research Laboratory, Department of Rheumatology and Inflammation Research, Sahlgrenska Academy,
University of Gothenburg, Gothenburg, Sweden.
4. Division of Clinical Microbiology, Department of Clinical and Experimental Medicine, Lab1, level 10, Faculty of Health Sciences, Linköping University, Linköping, Sweden.
5. Division of Infectious Diseases, Department of Medical and Health Sciences, University of Linköping, Linköping, Sweden.
6. Division of Radiology, Department of Medical and Health Sciences, University of Linköping, Linköping, Sweden.
7. Department of Clinical Microbiology and Infectious diseases, Kalmar County Hospital, Kalmar, Sweden.
Background: The role of vitamin D supplementation as adjuvant treatment of tuberculosis (TB) has lately attracted increasing interest. Our aim was to investigate the capacity of alveolar macrophages (AMs) from patients with or without exposure to TB to control intracellular growth of virulent Mycobacterium tuberculosis (Mtb).
Methods: AMs were freshly harvested from the bronchoalveolar lavage fluid of 7 patients with a history of TB (4 patients with previous TB and 3 patients with current TB) and 4 non-TB subjects. The H37Rv strain, genetically modified to express Vibrio harveyi luciferase, was used to determine the growth of Mtb by luminometry in the AMs from study subjects. Cytokine levels in culture supernatants were determined using a flow cytometry-based bead array technique.
Results: AMs from patients with a TB history were less efficient in restricting Mtb growth. Stimulation with 100 nM1, 25-dihydroxyvitamin D (1,25D3) did not significantly influence the capacity of AMs from any study subjects to control the infection. Out of the cytokines evaluated (TNF-α, IL-1β, IL-10 and IL-12p40) only TNF-α demonstrated detectable levels in culture supernatants, but did not respond to stimulation with 1,25D3.
Conclusions: We conclude that AMs of TB-patients show reduced ability to control mycobacterial growth in vitro, and, that AMs in this pilot study do no respond to 1, 25D3-stimulation. The former observation supports the concept that innate immunity is crucial for the control of TB infection.
Keywords: Alveolar macrophages, bronchoalveolar lavage, cytokines, H37Rv, tuberculosis
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