Abstract
Cell biology differs between traditional cell culture and 3-dimensional (3-D) systems, and is modulated by the extracellular matrix. Experimentation in 3-D presents challenges, especially with virulent pathogens. Mycobacterium tuberculosis (Mtb) kills more humans than any other infection and is characterised by a spatially organised immune response and extracellular matrix remodelling. We developed a 3-D system incorporating virulent mycobacteria, primary human blood mononuclear cells and collagen-alginate matrix to dissect the host-pathogen interaction. Infection in 3-D led to greater cellular survival and permitted longitudinal analysis over 21 days. Key features of human tuberculosis develop, and extracellular matrix integrity favours the host over the pathogen. We optimised multiparameter readouts to study emerging therapeutic interventions: cytokine supplementation, host-directed therapy and immunoaugmentation. Each intervention modulates the host-pathogen interaction, but has both beneficial and harmful effects. This methodology has wide applicability to investigate infectious, inflammatory and neoplastic diseases and develop novel drug regimes and vaccination approaches.
Keywords:
Mycobacterium tuberculosis; bioengineering; extracellular matrix; human; immunology; infectious disease; microbiology; modelling.
MeSH terms
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Alginates / chemistry
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Antigens, Bacterial / pharmacology
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Bacterial Proteins / pharmacology
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Chemokine CCL2 / biosynthesis
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Chemokine CCL2 / metabolism
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Chemokine CXCL10 / biosynthesis
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Chemokine CXCL10 / metabolism
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Coculture Techniques
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Collagen / chemistry
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Dinoprostone / pharmacology
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Extracellular Matrix / chemistry
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Extracellular Matrix / drug effects
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Extracellular Matrix / immunology
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Gene Expression Regulation
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Glucuronic Acid / chemistry
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Granulocyte-Macrophage Colony-Stimulating Factor / biosynthesis
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Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
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Hexuronic Acids / chemistry
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Host-Pathogen Interactions / drug effects*
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Host-Pathogen Interactions / immunology
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Humans
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Interleukin-12 / biosynthesis
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Interleukin-12 / metabolism
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Interleukin-1beta / biosynthesis
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Interleukin-1beta / metabolism
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Leukocytes, Mononuclear / drug effects*
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Leukocytes, Mononuclear / immunology
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Leukocytes, Mononuclear / microbiology
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Microspheres
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Models, Biological*
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Mycobacterium tuberculosis / drug effects
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Mycobacterium tuberculosis / pathogenicity*
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Mycobacterium tuberculosis / physiology
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Spheroids, Cellular / drug effects*
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Spheroids, Cellular / immunology
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Spheroids, Cellular / microbiology
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Virulence
Substances
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Alginates
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Antigens, Bacterial
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Bacterial Proteins
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CCL2 protein, human
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CFP-10 protein, Mycobacterium tuberculosis
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CXCL10 protein, human
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Chemokine CCL2
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Chemokine CXCL10
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ESAT-6 protein, Mycobacterium tuberculosis
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Hexuronic Acids
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IL1B protein, human
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Interleukin-1beta
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Interleukin-12
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Granulocyte-Macrophage Colony-Stimulating Factor
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Glucuronic Acid
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Collagen
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Dinoprostone