title: Stereological analysis of bacterial load and lung lesions in nonhuman primates (rhesus macaques) experimentally infected with Mycobacterium tuberculosis.
abstract: Infection with Mycobacterium tuberculosis primarily produces a multifocal distribution of pulmonary granulomas in which the pathogen resides. Accordingly, quantitative assessment of the bacterial load and pathology is a substantial challenge in tuberculosis. Such assessments are critical for studies of the pathogenesis and for the development of vaccines and drugs in animal models of experimental M. tuberculosis infection. Stereology enables unbiased quantitation of three-dimensional objects from two-dimensional sections and thus is suited to quantify histological lesions. We have developed a protocol for stereological analysis of the lung in rhesus macaques inoculated with a pathogenic clinical strain of M. tuberculosis (Erdman strain). These animals exhibit a pattern of infection and tuberculosis similar to that of naturally infected humans. Conditions were optimized for collecting lung samples in a nonbiased, random manner. Bacterial load in these samples was assessed by a standard plating assay, and granulomas were graded and enumerated microscopically. Stereological analysis provided quantitative data that supported a significant correlation between bacterial load and lung granulomas. Thus this stereological approach enables a quantitative, statistically valid analysis of the impact of M. tuberculosis infection in the lung and will serve as an essential tool for objectively comparing the efficacy of drugs and vaccines.
authors.affiliation: Center for Comparative Medicine, Davis, CA 95616, USA. paluciw@ucdavis.edu
authors.forename: Paul A
authors.initials: PA
authors.lastname: Luciw
authors.forename: Karen L
authors.initials: KL
authors.lastname: Oslund
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Yang
authors.forename: Lourdes
authors.initials: L
authors.lastname: Adamson
authors.forename: Resmi
authors.initials: R
authors.lastname: Ravindran
authors.forename: Don R
authors.initials: DR
authors.lastname: Canfield
authors.forename: Ross
authors.initials: R
authors.lastname: Tarara
authors.forename: Linda
authors.initials: L
authors.lastname: Hirst
authors.forename: Miles
authors.initials: M
authors.lastname: Christensen
authors.forename: Nicholas W
authors.initials: NW
authors.lastname: Lerche
authors.forename: Heather
authors.initials: H
authors.lastname: Offenstein
authors.forename: David
authors.initials: D
authors.lastname: Lewinsohn
authors.forename: Frank
authors.initials: F
authors.lastname: Ventimiglia
authors.forename: Laurie
authors.initials: L
authors.lastname: Brignolo
authors.forename: Erik R
authors.initials: ER
authors.lastname: Wisner
authors.forename: Dallas M
authors.initials: DM
authors.lastname: Hyde
mesh_terms: D000818:Animals; D058491:Bacterial Load; D001999:Bronchoscopy; D004195:Disease Models, Animal; D004801:Eosine Yellowish-(YS) / Q000032:analysis; D015769:Granuloma, Respiratory Tract / Q000150:complications / Q000382:microbiology / Q000473:pathology*; D006416:Hematoxylin / Q000032:analysis; D006801:Humans; D007442:Intubation, Intratracheal; D008168:Lung / Q000382:microbiology / Q000473:pathology*; D008253:Macaca mulatta; D008297:Male; D008853:Microscopy; D009169:Mycobacterium tuberculosis / Q000254:growth & development*; D009929:Organ Size; D012720:Severity of Illness Index; D014020:Tissue Extracts / Q000032:analysis; D014397:Tuberculosis, Pulmonary / Q000150:complications / Q000382:microbiology / Q000473:pathology*
chemical_list: D014020:Tissue Extracts; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin