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Primary macrophages and J774 cells respond differently to infection with Mycobacterium tuberculosis - PubMed

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Primary macrophages and J774 cells respond differently to infection with Mycobacterium tuberculosis

Nuria Andreu et al. Sci Rep. 2017.

Abstract

Macrophages play an essential role in the early immune response to Mycobacterium tuberculosis and are the cell type preferentially infected in vivo. Primary macrophages and macrophage-like cell lines are commonly used as infection models, although the physiological relevance of cell lines, particularly for host-pathogen interaction studies, is debatable. Here we use high-throughput RNA-sequencing to analyse transcriptome dynamics of two macrophage models in response to M. tuberculosis infection. Specifically, we study the early response of bone marrow-derived mouse macrophages and cell line J774 to infection with live and γ-irradiated (killed) M. tuberculosis. We show that infection with live bacilli specifically alters the expression of host genes such as Rsad2, Ifit1/2/3 and Rig-I, whose potential roles in resistance to M. tuberculosis infection have not yet been investigated. In addition, the response of primary macrophages is faster and more intense than that of J774 cells in terms of number of differentially expressed genes and magnitude of induction/repression. Our results point to potentially novel processes leading to immune containment early during M. tuberculosis infection, and support the idea that important differences exist between primary macrophages and cell lines, which should be taken into account when choosing a macrophage model to study host-pathogen interactions.

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Conflict of interest statement

The authors declare no competing financial interests.

Figures

Figure 1
Figure 1. Number of differentially expressed genes (DEGs).

(a) In BMDMs, and (b) in J774 infected with live M. tuberculosis or stimulated with γ-irradiated M. tuberculosis, relative to their time-matched uninfected controls, at 4 and 24 hpi (FDR < 0.05). The colour shading indicates the fold change in gene expression. The number of DEGs is indicated.

Figure 2
Figure 2. Comparison of the response of BMDMs and J774 to infection with live M. tuberculosis.

Scatterplots comparing the Log2 fold change (FC) of all the differentially expressed genes (DEGs) in BMDMs and J774 at 4 hpi (a) and 24 hpi (d). Grey dots represent genes differentially expressed in one macrophage type only. Points in shades of red represent genes differentially expressed in both data sets in the same direction. Points in shades of green represent genes differentially expressed in opposite direction. Those showing FC < |2| in either/both condition(s) are shown in light colour and those with FC > |2| in both conditions are shown in dark. The number of genes are indicated for each quadrant. (b) and (e) Venn diagrams comparing DEGs in BMDMs and J774 at 4 hpi and 24 hpi, respectively. (c) and (f) Line graphs showing the variation of the Log2FC for each DEG in common between BMDMs and J774 at 4 hpi (c) and 24 hpi (f). Lines are coloured according to the Log2FC in BMDMs (Log2FC > 4 red, 2 < Log2FC < 4 orange, 1 < Log2FC < 2 yellow, −1 > Log2FC > −2 green, −2 > Log2FC blue).

Figure 3
Figure 3. Top enriched pathways in macrophages infected with live M. tuberculosis.

Data were analysed through the use of QIAGEN’s Ingenuity® Pathway Analysis (IPA®, QIAGEN Redwood City,

www.qiagen.com/ingenuity

). Pathways are ranked according to (a) the enrichment score (Fisher’s exact test P-value), and (b) the Z-score that predicts activation/repression.

Figure 4
Figure 4. Comparison of the response of BMDMs and J774 to infection with live bacteria and stimulation with dead M. tuberculosis.

Scatterplots comparing the Log2 fold change (FC) of all the differentially expressed genes (DEGs) in BMDMs at 4 hpi (a) and 24 hpi (b), and J774 at 4 hpi (c) and 24 hpi (d). Grey dots represent genes differentially expressed in one condition only. Points in shades of red represent genes differentially expressed in both data sets in the same direction. Points in shades of green represent genes differentially expressed in opposite direction. Those showing FC < |2| in either/both condition(s) are shown in light colour and those with FC > |2| in both conditions are shown in dark. The number of genes are indicated for each quadrant. A Venn diagram showing the number of DEGs in macrophages infected with live bacteria and/or stimulated with dead bacteria is shown for each macrophage type and time point.

Figure 5
Figure 5. Top enriched pathways in macrophages infected with live bacteria and stimulated with dead M. tuberculosis.

(a) BMDMs, and (b) J774 cells. Data were analysed through the use of QIAGEN’s Ingenuity® Pathway Analysis (IPA®, QIAGEN Redwood City,

www.qiagen.com/ingenuity

). Pathways are ranked according to the Z-score that predicts activation/repression.

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