title: Multiple routes of complement activation by Mycobacterium bovis BCG.
abstract: Mycobacterium tuberculosis is the leading cause of infectious disease in humans in the world. It evades the host immune system by being phagocytosed by macrophages and residing intracellularly. Complement-dependent opsonisation of extracellular mycobacteria may assist them to enter macrophages. This work examines in detail the mechanisms of complement activation by whole mycobacteria using Mycobacterium bovis BCG as a model organism. M. bovis BCG directly activates the classical, lectin and alternative pathways, resulting in fixation of C3b onto macromolecules of the mycobacterial surface. Investigation into the classical pathway has shown direct binding of human C1q to whole mycobacteria in the absence of antibodies. Most human sera contain IgG and IgM-anti-(M. bovis BCG), and pre-incubation with human immunoglobulin enhances C1q binding to the bacteria. Therefore classical pathway activation is both antibody-independent and dependent. The bacteria also activate the alternative pathway in an antibody-independent manner, but Factor H also binds, suggesting some regulation of amplification by this pathway. For the lectin pathway we have demonstrated direct binding of both MBL and L-ficolin from human serum to whole mycobacteria and subsequent MASP2 activation. H-ficolin binding was not observed. No M. bovis BCG cell surface or secreted protease appears likely to influence complement activation. Together, these data provide a more detailed analysis of the mechanisms by which M. bovis BCG interacts with the complement system.
authors.affiliation: MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, Oxford, UK. maria.carroll@bioch.ox.ac.uk
authors.forename: Maria V
authors.initials: MV
authors.lastname: Carroll
authors.forename: Nathan
authors.initials: N
authors.lastname: Lack
authors.forename: Edith
authors.initials: E
authors.lastname: Sim
authors.forename: Anders
authors.initials: A
authors.lastname: Krarup
authors.forename: Robert B
authors.initials: RB
authors.lastname: Sim
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology*; D003167:Complement Activation / Q000276:immunology*; D015922:Complement C1q / Q000276:immunology; D003179:Complement C3b / Q000276:immunology; D017242:Complement Factor H / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007075:Immunoglobulin M / Q000276:immunology*; D037102:Lectins / Q000276:immunology; D008264:Macrophages / Q000276:immunology*; D037601:Mannose-Binding Lectin / Q000276:immunology; D050606:Mannose-Binding Protein-Associated Serine Proteases / Q000276:immunology; D009163:Mycobacterium bovis / Q000276:immunology*; D000097604:Ficolins
chemical_list: D000907:Antibodies, Bacterial; D015922:Complement C1q; D003179:Complement C3b; D017242:Complement Factor H; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D037102:Lectins; D037601:Mannose-Binding Lectin; D050606:Mannose-Binding Protein-Associated Serine Proteases; D000097604:Ficolins; C105296:MASP2 protein, human