Quantitative expression of C-type lectin receptors in humans and mice - PubMed (original) (raw)

Comparative Study

. 2012;13(8):10113-10131.

doi: 10.3390/ijms130810113. Epub 2012 Aug 14.

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Comparative Study

Quantitative expression of C-type lectin receptors in humans and mice

Maciej Lech et al. Int J Mol Sci. 2012.

Abstract

C-type lectin receptors and their adaptor molecules are involved in the recognition of glycosylated self-antigens and pathogens. However, little is known about the species- and organ-specific expression profiles of these molecules. We therefore determined the mRNA expression levels of Dectin-1, MR1, MR2, DC-SIGN, Syk, Card-9, Bcl-10, Malt-1, Src, Dec-205, Galectin-1, Tim-3, Trem-1, and DAP-12 in 11 solid organs of human and mice. Mouse organs revealed lower mRNA levels of most molecules compared to spleen. However, Dec-205 and Galectin-1 in thymus, Src in brain, MR2, Card-9, Bcl-10, Src, and Dec-205 in small intestine, MR2, Bcl-10, Src, Galectin-1 in kidney, and Src and Galectin-1 in muscle were at least 2-fold higher expressed compared to spleen. Human lung, liver and heart expressed higher mRNA levels of most genes compared to spleen. Dectin-1, MR1, Syk and Trem-1 mRNA were strongly up-regulated upon ischemia-reperfusion injury in murine kidney. Tim3, DAP-12, Card-9, DC-SIGN and MR2 were further up-regulated during renal fibrosis. Murine kidney showed higher DAP-12, Syk, Card-9 and Dectin-1 mRNA expression during the progression of lupus nephritis. Thus, the organ-, and species-specific expression of C-type lectin receptors is different between mice and humans which must be considered in the interpretation of related studies.

Keywords: dendritic cells; infection; inflammation; innate immunity; macrophages; pattern recognition receptors.

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Figures

Figure 1

Figure 1

(A) C-type lectin receptors (CLR) mRNA expression in adult human tissues. Quantitative real-time PCR analysis was performed on pre-normalized cDNAs derived from poly(A)-selected DNase-treated RNAs purified from pools of healthy human tissues as described in methods; (B) CLR mRNA expression in adult mouse tissues. Real-time PCR was performed on pooled cDNAs derived from five adult six to eight week old C57BL/6 mice as described in methods. Biological replicates were quantified in triplicates and normalized to the respective GAPDH mRNA level. The results in the table are expressed relative to the respective expression level of each transcript in spleen. In the table, red shades indicate higher and green shades indicate lower mRNA levels as compared to the respective mRNA levels in spleen. The spleen mRNA levels are illustrated in the histogram on top of the table. Data in B represents means ± SEM. * indicated statistical significance p < 0.05.

Figure 1

Figure 1

(A) C-type lectin receptors (CLR) mRNA expression in adult human tissues. Quantitative real-time PCR analysis was performed on pre-normalized cDNAs derived from poly(A)-selected DNase-treated RNAs purified from pools of healthy human tissues as described in methods; (B) CLR mRNA expression in adult mouse tissues. Real-time PCR was performed on pooled cDNAs derived from five adult six to eight week old C57BL/6 mice as described in methods. Biological replicates were quantified in triplicates and normalized to the respective GAPDH mRNA level. The results in the table are expressed relative to the respective expression level of each transcript in spleen. In the table, red shades indicate higher and green shades indicate lower mRNA levels as compared to the respective mRNA levels in spleen. The spleen mRNA levels are illustrated in the histogram on top of the table. Data in B represents means ± SEM. * indicated statistical significance p < 0.05.

Figure 2

Figure 2

CLR mRNA expression in adult human and mouse tissues. The respective relative human (open bars) and murine (black bars) CLRs mRNA levels from figure 1A,B are illustrated. The _x_-axis marks a ratio of 1, hence, positive values indicate stronger expression in humans, negative values indicate stronger expression in mice. The _y_-axis marks the fold-change in each direction. Note that the scale of the _y_-axis is different for each organ.

Figure 2

Figure 2

CLR mRNA expression in adult human and mouse tissues. The respective relative human (open bars) and murine (black bars) CLRs mRNA levels from figure 1A,B are illustrated. The _x_-axis marks a ratio of 1, hence, positive values indicate stronger expression in humans, negative values indicate stronger expression in mice. The _y_-axis marks the fold-change in each direction. Note that the scale of the _y_-axis is different for each organ.

Figure 3

Figure 3

(A) CLRs mRNA expression in mouse kidney upon induction of IRI; (B) CLRs mRNA expression in mouse kidney upon induction of UUO; (C) CLRs mRNA expression in mouse kidney upon induction of lupus nephritis (LN). Real-time PCR was performed on cDNAs derived from murine kidney tissue from five to ten C57BL/6 mice that underwent ischemia reperfusion injury (IRI) (A), unilateral ureteral obstruction (UUO) (B) or progressive systemic autoimmunity with lupus nephritis in MRLlpr mice (C) as described in methods. Biological replicates were quantified and normalized to 18s rRNA level. The results in the table are expressed relative to the respective expression level of each transcript in sham operated kidney (A, B) or six week old MRLlpr mice (C). In the table, red shades indicate higher and green shades indicate lower mRNA levels as compared to the respective mRNA levels in spleen. Basal mRNA levels in control kidney are illustrated in the histogram below the table. Data in histograms represents means ± SEM. * In the tables indicates statistical significance p < 0.05.

Figure 4

Figure 4

Renal leukocyte recruitment upon renal ischemia-reperfusion injury. Kidneys were obtained from all IRI mice at various time intervals as indicated. Immunostaining for macrophages and T-cells was performed on renal sections as described in material and methods. Data represents positive stained surface in percent and are shown as mean ± SEM from 8 to 12 mice (at least 25 hpf) of each group. F4/80, DAP-12 and Syk immunostainings were performed on paraffin embedded renal sections, obtained from mice 10 days after IRI, as described in material and methods. Data shows most representative results from each group. Data in histograms represents means ± SEM. ** Indicates statistical significance p < 0.01, *** indicates statistical significance p < 0.005.

Figure 5

Figure 5

Renal leukocyte recruitment upon unilateral ureteral obstruction. Kidneys were obtained from mice after UUO at various time intervals as indicated. Immunostaining for macrophages and T-cells was performed on renal sections as described in material and methods. Data represents positive stained surface in percent and are shown as mean ± SEM from 8 to 12 mice (at least 25 hpf) of each group. F4/80, DAP-12 and Syk immunostainings were performed on paraffin embedded renal sections, obtained from mice 10 days after UUO, as described in material and methods. Data shows most representative results from each group. Data in histograms represents means ± SEM. ** Indicates statistical significance p < 0.01, *** indicates statistical significance p < 0.005.

Figure 6

Figure 6

Renal leukocyte recruitment during systemic lupus of MRLpr mice. Kidneys were obtained from MRLlpr mice at various time intervals as indicated. Immunostaining for macrophages and T-cells was performed on renal sections as described in material and methods. Data represents positive stained surface in percent and are shown as mean ± SEM from 8 to 12 mice (at least 25 hpf) of each group. F4/80, DAP-12 and Syk immunostainings were performed on paraffin embedded renal sections as described in material and methods. Kidneys were obtained from 126 days old MRLlpr mice. Glomeruli are indicated by encircling them. Data shows most representative results from each group. Data in histograms represents means ± SEM. * Indicates statistical significance p < 0.05.

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