The human host’s innate immune responses involved in tuberculosis (TB) are central to the recognition and control of the infection (1). Toll-like receptors (TLRs) are an important part of this process, although their role, especially in humans is not yet fully defined (2-3). We aim to characterise TLR expression and function in patients with TB and examine the effect of anti-tuberculous therapy.
Patients with active TB had blood samples taken during antimicrobial therapy (baseline, 1 month, 2 months and end-of-treatment). Active TB was defined by positive culture, PCR or if presumed clinically. TLR expression (TLR2,4 and 7/8) on peripheral blood mononuclear cells (PBMCs) was measured by flow cytometry. Cytokine levels (TNFα, IL-6) using ELISA were measured following stimulation of PBMCs with TLR-ligands to determine function.
Preliminary data of the first 14 patients with active TB is presented here.
TLR expression:
There was a significant decrease in TLR4 expression (p value=0.0391) and a trend to decreased TLR2 expression (p=0.0547) on CD14+ monocytes after treatment. TLR7 expression increased in these cells when comparing early and end-of treatment (p=0.0391). Interestingly, there was a trend to an increase in TLR7 expression when comparing early and end-of treatment samples in NKT cells (CD3+, CD56+) and CD56dim NK cells (p=0.0547-0.0742 respectively).
Stimulated Cytokine measurements:
TNFα levels increased significantly between baseline, early and end-of treatment following stimulation with TLR9 ligand CpG (p=0.021 – p=0.0498), and without stimulation (p=0.0156 - p=0.0313). IL-6 levels were also significantly increased between baseline and end-of treatment (p=0.0039) following stimulation with TLR9 ligand.
Conclusions:
The differences in TLR expression and cytokine levels in this pilot study highlight the potential of utilising these to form a better understanding of longitudinal changes following anti-microbial therapy in active TB. Our findings suggest that measurement of innate immune responses may provide a means of measuring treatment responses.
- (1) Gerhard W et al (2011). Nature Reviews Immunology 11(May ): 343-354.
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(2) Hiroyuki S. et al, (2011). Clinical and developmental immunology 2011(Article ID 347594).
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(3) Druszczynska M et al (2011). Tuberculosis:Immunology, Cell Biology and Novel Vaccination Strategies. Vancouver, Canada.