Stereological analysis of bacterial load and lung lesions in nonhuman primates (rhesus macaques) experimentally infected with Mycobacterium tuberculosis

Paul A. Luciw, Karen L. Oslund, Xiao Wei Yang, Lourdes Adamson, Resmi Ravindran, Don R. Canfield, Ross Tarara, Linda Hirst, Miles Christensen, Nicholas W. Lerche, Heather Offenstein, David Lewinsohn, Frank Ventimiglia, Laurie Brignolo, Erik R. Wisner, Dallas M. Hyde

Research output: Contribution to journalArticle

22 Citations (Scopus)

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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume301
Issue number5
DOIs
StatePublished - Nov 2011

Fingerprint

Bacterial Load
Macaca mulatta
Mycobacterium tuberculosis
Primates
Lung
Granuloma
Mycobacterium Infections
Tuberculosis
Vaccines
Infection
Pharmaceutical Preparations
Animal Models
Pathology

Keywords

  • Granuloma
  • Morphometry
  • Quantitative pathology

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology
  • Physiology

Cite this

Stereological analysis of bacterial load and lung lesions in nonhuman primates (rhesus macaques) experimentally infected with Mycobacterium tuberculosis. / Luciw, Paul A.; Oslund, Karen L.; Yang, Xiao Wei; Adamson, Lourdes; Ravindran, Resmi; Canfield, Don R.; Tarara, Ross; Hirst, Linda; Christensen, Miles; Lerche, Nicholas W.; Offenstein, Heather; Lewinsohn, David; Ventimiglia, Frank; Brignolo, Laurie; Wisner, Erik R.; Hyde, Dallas M.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 301, No. 5, 11.2011.

Research output: Contribution to journalArticle

Luciw, PA, Oslund, KL, Yang, XW, Adamson, L, Ravindran, R, Canfield, DR, Tarara, R, Hirst, L, Christensen, M, Lerche, NW, Offenstein, H, Lewinsohn, D, Ventimiglia, F, Brignolo, L, Wisner, ER & Hyde, DM 2011, 'Stereological analysis of bacterial load and lung lesions in nonhuman primates (rhesus macaques) experimentally infected with Mycobacterium tuberculosis', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 301, no. 5. https://doi.org/10.1152/ajplung.00120.2011
Luciw, Paul A. ; Oslund, Karen L. ; Yang, Xiao Wei ; Adamson, Lourdes ; Ravindran, Resmi ; Canfield, Don R. ; Tarara, Ross ; Hirst, Linda ; Christensen, Miles ; Lerche, Nicholas W. ; Offenstein, Heather ; Lewinsohn, David ; Ventimiglia, Frank ; Brignolo, Laurie ; Wisner, Erik R. ; Hyde, Dallas M. / Stereological analysis of bacterial load and lung lesions in nonhuman primates (rhesus macaques) experimentally infected with Mycobacterium tuberculosis. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2011 ; Vol. 301, No. 5.
@article{d30b4bced92249a99e0808ce41f5e37e,
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.",
keywords = "Granuloma, Morphometry, Quantitative pathology",
author = "Luciw, {Paul A.} and Oslund, {Karen L.} and Yang, {Xiao Wei} and Lourdes Adamson and Resmi Ravindran and Canfield, {Don R.} and Ross Tarara and Linda Hirst and Miles Christensen and Lerche, {Nicholas W.} and Heather Offenstein and David Lewinsohn and Frank Ventimiglia and Laurie Brignolo and Wisner, {Erik R.} and Hyde, {Dallas M.}",
year = "2011",
month = "11",
doi = "10.1152/ajplung.00120.2011",
language = "English (US)",
volume = "301",
journal = "American Journal of Physiology - Renal Fluid and Electrolyte Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "5",

}

TY - JOUR

T1 - Stereological analysis of bacterial load and lung lesions in nonhuman primates (rhesus macaques) experimentally infected with Mycobacterium tuberculosis

AU - Luciw, Paul A.

AU - Oslund, Karen L.

AU - Yang, Xiao Wei

AU - Adamson, Lourdes

AU - Ravindran, Resmi

AU - Canfield, Don R.

AU - Tarara, Ross

AU - Hirst, Linda

AU - Christensen, Miles

AU - Lerche, Nicholas W.

AU - Offenstein, Heather

AU - Lewinsohn, David

AU - Ventimiglia, Frank

AU - Brignolo, Laurie

AU - Wisner, Erik R.

AU - Hyde, Dallas M.

PY - 2011/11

Y1 - 2011/11

N2 - 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.

AB - 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.

KW - Granuloma

KW - Morphometry

KW - Quantitative pathology

UR - http://www.scopus.com/inward/record.url?scp=80055108676&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80055108676&partnerID=8YFLogxK

U2 - 10.1152/ajplung.00120.2011

DO - 10.1152/ajplung.00120.2011

M3 - Article

VL - 301

JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

SN - 1931-857X

IS - 5

ER -