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Isolation, Transfection, and Culture of Primary Human Monocytes
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Isolation, Transfection, and Culture of Primary Human Monocytes

Published on: December 16, 2019

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Human blood monocytes support persistence, but not replication of the intracellular pathogen C. pneumoniae

Tanja Buchacher1, Herbert Wiesinger-Mayr2, Klemens Vierlinger3

  • 1Christian Doppler Laboratory for Innovative Therapy Approaches in Sepsis, Danube University Krems, Krems, Austria. tanja.buchacher@donau-uni.ac.at.

BMC Immunology
|December 10, 2014
PubMed
Abstract

Insights

Infection with Chlamydia pneumoniae alters monocyte cytokine and chemokine profiles differently than LPS stimulation. Novel methods like DNA microarrays and Raman microspectroscopy effectively detect intracellular pathogen infection in monocytes.

Area of Science:

  • Immunology
  • Cell Biology
  • Microbiology

Background:

  • Intracellular pathogens employ immune evasion strategies.
  • Human monocytes are key immune cells susceptible to intracellular pathogens.
  • Chlamydia pneumoniae (C. pneumoniae) is an important intracellular pathogen.

Purpose of the Study:

  • To investigate the impact of C. pneumoniae infection on human monocyte cytokine and chemokine profiles.
  • To compare the monocyte response to C. pneumoniae infection versus lipopolysaccharide (LPS) stimulation.
  • To evaluate novel diagnostic tools for intracellular pathogen detection in monocytes.

Main Methods:

  • Human peripheral blood monocytes were infected with C. pneumoniae.
  • Infection was confirmed using immunofluorescence, real-time PCR, and DNA microarray.
  • Molecular changes were analyzed using Raman microspectroscopy.
  • Cytokine and chemokine profiles were assessed following stimulation.

Main Results:

  • C. pneumoniae infection was confirmed, though replication was not evident via PCR.
  • Raman microspectroscopy showed distinct molecular fingerprints in infected monocytes, primarily altered lipid content.
  • Both C. pneumoniae and LPS induced TNF-α and IL-6.
  • C. pneumoniae induced significantly higher levels of IL-1β, IL-12p40, and IL-12p70.
  • C. pneumoniae triggered the release of MCP-1, MIP-1α, MIP-1β, and CXCL-8 chemokines.

Conclusions:

  • Intracellular pathogen infection of monocytes elicits a unique cytokine and chemokine response compared to extracellular stimuli like LPS.
  • Oligonucleotide DNA microarrays and Raman microspectroscopy are valuable tools for identifying monocyte infection by intracellular pathogens.

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