Different transcriptional profiles of RAW264.7 infected with Mycobacterium tuberculosis H37Rv and BCG identified via deep sequencing

PLoS One. 2012;7(12):e51988. doi: 10.1371/journal.pone.0051988. Epub 2012 Dec 19.

Abstract

Background: The Mycobacterium tuberculosis H37Rv and BCG effects on the host cell transcriptional profile consider a main research point. In the present study the transcriptome profiling analysis of RAW264.7 either infected with Mycobacterium tuberculosis H37Rv or BCG have been reported using Solexa/Illumina digital gene expression (DGE).

Results: The DGE analysis showed 1,917 different expressed genes between the BCG and H37Rv group. In addition, approximately 5% of the transcripts appeared to be predicted genes that have never been described before. KEGG Orthology (KO) annotations showed more than 71% of these transcripts are possibly involved in approximately 210 known metabolic or signaling pathways. The gene of the 28 pathways about pathogen recognition receptors and Mycobacterium tuberculosis interaction with macrophages were analyzed using the CLUSTER 3.0 available, the Tree View tool and Gene Orthology (GO). Some genes were randomly selected to confirm their altered expression levels by quantitative real-time PCR (qRT-PCR).

Conclusion: The present study used DGE from pathogen recognition receptors and Mycobacterium tuberculosis interaction with macrophages to understand the interplay between Mycobacterium tuberculosis and RAW264.7. Meanwhile find some important host protein which was affected by Mycobacterium tuberculosis to provide evidence for the further improvement of the present efficacy of existing Mycobacterium tuberculosis therapy and vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • BCG Vaccine* / immunology
  • Cell Line
  • Cluster Analysis
  • Expressed Sequence Tags
  • Gene Expression Profiling*
  • Gene Expression Regulation
  • Gene Library
  • High-Throughput Nucleotide Sequencing
  • Host-Pathogen Interactions / genetics
  • Host-Pathogen Interactions / immunology
  • Macrophages / immunology
  • Macrophages / metabolism*
  • Macrophages / microbiology*
  • Mice
  • Molecular Sequence Annotation
  • Mycobacterium tuberculosis* / immunology
  • Receptors, Pattern Recognition / genetics
  • Receptors, Pattern Recognition / immunology
  • Reproducibility of Results
  • Stress, Physiological / genetics
  • Stress, Physiological / immunology

Substances

  • BCG Vaccine
  • Receptors, Pattern Recognition

Grants and funding

This work was supported by National Science and Technology Major Project (2009ZX08009-163B, http://www.nmp.gov.cn/), National Natural Science Foundation of China (Key Program, No. 31101813, http://www.nsfc.gov.cn) and Jilin Province Science and Technology Developing Project (20076013). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.