A comparative analysis of gene expression patterns and cell phenotypes between cervical and peripheral blood mononuclear cells

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Abstract

Studies of the immunological environment in the female genital tract (FGT) are critical for the development of vaccines or microbicides to halt the spread of sexually transmitted infections. Challenges arise due to the difficulties of sampling from this site, and the majority of studies have been conducted utilising peripheral blood mononuclear cells. Identifying functional differences between immune cells of the FGT and peripheral blood would aid in our understanding of mucosal immunology. We compared the gene expression profile of mononuclear cells at these two sites. Messenger RNA expression analysis was performed using gene expression arrays on matched cervical mononuclear cells and peripheral blood mononuclear cells. Further cellular phenotyping was done by 10 colour flow cytometry. Of the 22,185 genes expressed by these samples, 5345 genes were significantly differentially expressed between the cell populations. Most differences can be explained by significantly lower levels of T and B cells and higher levels of macrophages and dendritic cells in the FGT compared with peripheral blood. Several immunologically relevant pathways such as apoptosis and innate immune signalling, and a variety of cytokines and cytokine receptors were differentially expressed. This study highlights the importance of the unique immunological environment of the FGT and identifies important differences between systemic and mucosal immune compartments. © 2009 Horton et al.

Figures

  • Figure 1. Overall gene expression in matched CMCs and PBMCs. CMCs and PBMCs cluster independently using unsupervised hierarchical clustering. Red indicates a relative increase in gene expression intensity and green represents a relative decrease in gene expression intensity in the sample. doi:10.1371/journal.pone.0008293.g001
  • Figure 2. Relative differences between the immune cells of PBMC and CMC populations. (A) Columns show mean of 5 individuals with SEM shown by vertical lines. (B) Relative gene expression values of phenotyping markers. Red indicates a relative increase in gene expression intensity and green represents a relative decrease in gene expression intensity. doi:10.1371/journal.pone.0008293.g002
  • Figure 3. Gene expression heatmaps of differentially expressed pathways. Significant pathways were identified by DAVID with EASE scores ,0.05. Red indicates a relative increase in gene expression intensity in the sample and green represents a relative decrease in gene expression intensity. Gene symbols are listed on the right of each heatmap. (A) represents all genes expressed by CMCs and PBMCs involved in the cytokinecytokine receptor interaction pathway, (B) represents genes expressed in the apoptosis pathway, and (C) represents all genes expressed by CMCs and PBMCs involved in TLR signalling. doi:10.1371/journal.pone.0008293.g003
  • Figure 4. Apoptosis pathway (modified from KEGG). CFLAR, denoted by *, is an important inhibitor of apoptosis and is significantly over expressed by CMCs by a factor of 2. doi:10.1371/journal.pone.0008293.g004
  • Figure 5. TLR signalling pathway (modified from KEGG). MYD88, denoted by *, is an important inhibitor of apoptosis and is significantly over expressed by CMCs by a factor of 2.39. doi:10.1371/journal.pone.0008293.g005
  • Figure 6. Gating scheme. Representative dot plots showing the gating scheme used to determine different mononuclear cell subtypes and via lineage negative gating obtain the Dendritic cell population of PBMCs (A) and CMCs (B). Cells were obtained from peripheral blood and cervical scrapings by ficoll separation and labelled with a cocktail of antibodies (CD3-AmCyan, CD14-Pacific Blue, CD19-PECy7, CD56-Alexa Fluor 700 and HLA DR- APCCy7) then 10 colour flow cytometry was performed on a LSR II. doi:10.1371/journal.pone.0008293.g006

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CITATION STYLE

APA

Horton, R. E., Kaefer, N., Songok, E., Guijon, F. B., Kettaf, N., Boucher, G., … Plummer, F. A. (2009). A comparative analysis of gene expression patterns and cell phenotypes between cervical and peripheral blood mononuclear cells. PLoS ONE, 4(12). https://doi.org/10.1371/journal.pone.0008293

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