title: Assessment of immunity to mycobacterial infection with luciferase reporter constructs.
abstract: Protective immunity to mycobacterial infection is incompletely understood but probably involves the coordinated interaction of multiple cell types and cytokines. With the aim of developing assays that might provide a surrogate measure of protective immunity, we have investigated the use of recombinant mycobacteria carrying luciferase reporter enzymes to assess the effectiveness of antimycobacterial immunity in model systems. Measurement of luminescence was shown to provide a rapid and simple alternative to the counting of CFU as a means of monitoring mycobacterial viability. We describe optimization of a luciferase reporter strain of Mycobacterium tuberculosis and demonstrate its application for the study of mycobacterial interactions with host cells in tissue culture and the rapid assessment of vaccine efficacy in a murine model.
authors.affiliation: Department of Infectious Diseases and Microbiology, Imperial College School of Medicine, St. Mary's Campus, London W2 1PG, United Kingdom. v.snewin@ic.ac.uk
authors.forename: V A
authors.initials: VA
authors.lastname: Snewin
authors.forename: M P
authors.initials: MP
authors.lastname: Gares
authors.forename: P O
authors.initials: PO
authors.lastname: Gaora
authors.forename: Z
authors.initials: Z
authors.lastname: Hasan
authors.forename: I N
authors.initials: IN
authors.lastname: Brown
authors.forename: D B
authors.initials: DB
authors.lastname: Young
mesh_terms: D000818:Animals; D001500:BCG Vaccine / Q000276:immunology; D002460:Cell Line; D004195:Disease Models, Animal; D005260:Female; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000276:immunology / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D014376:Tuberculosis / Q000276:immunology*; D014611:Vaccination
chemical_list: D001500:BCG Vaccine; D008156:Luciferases