Gene expression in the tuberculous granuloma: Analysis by laser capture microdissection and real-time PCR

Guofeng Zhu, Huifang Xiao, Vellore P. Mohan, Kathryn E. Tanaka, Sanjay Tyagi, Fred Tsen, Padmini Salgame, John Chan

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

We have assessed the kinetics of host gene expression in granulomas of mice infected with virulent Mycobacterium tuberculosis, using an approach that incorporates the laser capture microdissection (LCM) and real-time PCR technology in conjunction with a newly derived mathematical equation. The results have provided evidence indicating that conventional use of whole infected lungs to study granuloma-specific gene expression can yield data that may not genuinely reflect intralesional events. Significantly, the expression of nine host genes known to regulate the inflammatory response to M. tuberculosis, as determined by real-time PCR analysis of microdissected granuloma-derived cDNAs, was downregulated (up to 27-fold) at around the time when the rapid growth phase of the bacilli in the lungs of infected mice ends. This downregulation was masked when whole infected lungs were used for the studies. The data suggest that the host immune system can adjust and respond to, or can be modulated by specific physiological states of the tubercle bacillus in vivo. The LCM/real-time PCR-based system described in this study can be applied to safely and accurately evaluate gene expression in any lesions that can be microscopically visualized, including those contained in biohazardous tissues.

Original languageEnglish (US)
Pages (from-to)445-453
Number of pages9
JournalCellular Microbiology
Volume5
Issue number7
DOIs
StatePublished - Jul 1 2003

Fingerprint

Microdissection
Laser Capture Microdissection
Granuloma
Gene expression
Real-Time Polymerase Chain Reaction
Bacilli
Mycobacterium tuberculosis
Gene Expression
Lung
Bacillus
Lasers
Down-Regulation
Immune system
Immune System
Complementary DNA
Genes
Tissue
Technology
Kinetics
Growth

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Microbiology

Cite this

Gene expression in the tuberculous granuloma : Analysis by laser capture microdissection and real-time PCR. / Zhu, Guofeng; Xiao, Huifang; Mohan, Vellore P.; Tanaka, Kathryn E.; Tyagi, Sanjay; Tsen, Fred; Salgame, Padmini; Chan, John.

In: Cellular Microbiology, Vol. 5, No. 7, 01.07.2003, p. 445-453.

Research output: Contribution to journalArticle

Zhu, Guofeng ; Xiao, Huifang ; Mohan, Vellore P. ; Tanaka, Kathryn E. ; Tyagi, Sanjay ; Tsen, Fred ; Salgame, Padmini ; Chan, John. / Gene expression in the tuberculous granuloma : Analysis by laser capture microdissection and real-time PCR. In: Cellular Microbiology. 2003 ; Vol. 5, No. 7. pp. 445-453.
@article{2abf263002254b9e88527992626049f3,
title = "Gene expression in the tuberculous granuloma: Analysis by laser capture microdissection and real-time PCR",
abstract = "We have assessed the kinetics of host gene expression in granulomas of mice infected with virulent Mycobacterium tuberculosis, using an approach that incorporates the laser capture microdissection (LCM) and real-time PCR technology in conjunction with a newly derived mathematical equation. The results have provided evidence indicating that conventional use of whole infected lungs to study granuloma-specific gene expression can yield data that may not genuinely reflect intralesional events. Significantly, the expression of nine host genes known to regulate the inflammatory response to M. tuberculosis, as determined by real-time PCR analysis of microdissected granuloma-derived cDNAs, was downregulated (up to 27-fold) at around the time when the rapid growth phase of the bacilli in the lungs of infected mice ends. This downregulation was masked when whole infected lungs were used for the studies. The data suggest that the host immune system can adjust and respond to, or can be modulated by specific physiological states of the tubercle bacillus in vivo. The LCM/real-time PCR-based system described in this study can be applied to safely and accurately evaluate gene expression in any lesions that can be microscopically visualized, including those contained in biohazardous tissues.",
author = "Guofeng Zhu and Huifang Xiao and Mohan, {Vellore P.} and Tanaka, {Kathryn E.} and Sanjay Tyagi and Fred Tsen and Padmini Salgame and John Chan",
year = "2003",
month = "7",
day = "1",
doi = "10.1046/j.1462-5822.2003.00288.x",
language = "English (US)",
volume = "5",
pages = "445--453",
journal = "Cellular Microbiology",
issn = "1462-5814",
publisher = "Wiley-Blackwell",
number = "7",

}

TY - JOUR

T1 - Gene expression in the tuberculous granuloma

T2 - Analysis by laser capture microdissection and real-time PCR

AU - Zhu, Guofeng

AU - Xiao, Huifang

AU - Mohan, Vellore P.

AU - Tanaka, Kathryn E.

AU - Tyagi, Sanjay

AU - Tsen, Fred

AU - Salgame, Padmini

AU - Chan, John

PY - 2003/7/1

Y1 - 2003/7/1

N2 - We have assessed the kinetics of host gene expression in granulomas of mice infected with virulent Mycobacterium tuberculosis, using an approach that incorporates the laser capture microdissection (LCM) and real-time PCR technology in conjunction with a newly derived mathematical equation. The results have provided evidence indicating that conventional use of whole infected lungs to study granuloma-specific gene expression can yield data that may not genuinely reflect intralesional events. Significantly, the expression of nine host genes known to regulate the inflammatory response to M. tuberculosis, as determined by real-time PCR analysis of microdissected granuloma-derived cDNAs, was downregulated (up to 27-fold) at around the time when the rapid growth phase of the bacilli in the lungs of infected mice ends. This downregulation was masked when whole infected lungs were used for the studies. The data suggest that the host immune system can adjust and respond to, or can be modulated by specific physiological states of the tubercle bacillus in vivo. The LCM/real-time PCR-based system described in this study can be applied to safely and accurately evaluate gene expression in any lesions that can be microscopically visualized, including those contained in biohazardous tissues.

AB - We have assessed the kinetics of host gene expression in granulomas of mice infected with virulent Mycobacterium tuberculosis, using an approach that incorporates the laser capture microdissection (LCM) and real-time PCR technology in conjunction with a newly derived mathematical equation. The results have provided evidence indicating that conventional use of whole infected lungs to study granuloma-specific gene expression can yield data that may not genuinely reflect intralesional events. Significantly, the expression of nine host genes known to regulate the inflammatory response to M. tuberculosis, as determined by real-time PCR analysis of microdissected granuloma-derived cDNAs, was downregulated (up to 27-fold) at around the time when the rapid growth phase of the bacilli in the lungs of infected mice ends. This downregulation was masked when whole infected lungs were used for the studies. The data suggest that the host immune system can adjust and respond to, or can be modulated by specific physiological states of the tubercle bacillus in vivo. The LCM/real-time PCR-based system described in this study can be applied to safely and accurately evaluate gene expression in any lesions that can be microscopically visualized, including those contained in biohazardous tissues.

UR - http://www.scopus.com/inward/record.url?scp=0038049387&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038049387&partnerID=8YFLogxK

U2 - 10.1046/j.1462-5822.2003.00288.x

DO - 10.1046/j.1462-5822.2003.00288.x

M3 - Article

C2 - 12814435

AN - SCOPUS:0038049387

VL - 5

SP - 445

EP - 453

JO - Cellular Microbiology

JF - Cellular Microbiology

SN - 1462-5814

IS - 7

ER -