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Comparative Study
. 2006 Dec;7(8):667-79.
doi: 10.1038/sj.gene.6364345. Epub 2006 Oct 26.

Gene expression profiling in a mouse model for African trypanosomiasis

S Kierstein  1 , H Noyes, J Naessens, Y Nakamura, C Pritchard, J Gibson, S Kemp, A Brass
Affiliations
Comparative Study

Gene expression profiling in a mouse model for African trypanosomiasis

S Kierstein et al. Genes Immun. 2006 Dec.

Abstract

This study aimed to provide the foundation for an integrative approach to the identification of the mechanisms underlying the response to infection with Trypanosoma congolense, and to identify pathways that have previously been overlooked. We undertook a large-scale gene expression analysis study comparing susceptible A/J and more tolerant C57BL/6 mice. In an initial time course experiment, we monitored the development of parasitaemia and anaemia in every individual. Based on the kinetics of disease progression, we extracted total RNA from liver at days 0, 4, 7, 10 and 17 post infection and performed a microarray analysis. We identified 64 genes that were differentially expressed in the two strains in non-infected animals, of which nine genes remained largely unaffected by the disease. Gene expression profiling at stages of low, peak, clearance and recurrence of parasitaemia suggest that susceptibility is associated with high expression of genes coding for chemokines (e.g. Ccl24, Ccl27 and Cxcl13), complement components (C1q and C3) and interferon receptor alpha (Ifnar1). Additionally, susceptible A/J mice expressed higher levels of some potassium channel genes. In contrast, messenger RNA levels of a few immune response, metabolism and protease genes (e.g. Prss7 and Mmp13) were higher in the tolerant C57BL/6 strain as compared to A/J.

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Figures

Figure 1
Figure 1
Kinetics of T. congolense infection in A/J and C57BL/6 mice. Mice of the susceptible A/J and the resistant C57BL/6 stain were infected with T. congolense by i.p. injection of 1 × 104 parasites. Tail blood was collected every other day from each individual. (a) Parasites were counted from 3 μl of blood, diluted 1:200 in Alsevers solution, under the microscope using a haemocytometer. Average parasite counts are expressed as number of parasites per ml. A/J mice had significantly higher parasite counts at day 8 (t-test, P<0.0001), but both strains were able to reduce their trypanosome load by day 10 at which stage there was no significant difference between the two strains. However, the parasite numbers were again significantly higher in A/J mice at all following time points (two-way ANOVA, P<0.0001). (b) To determine blood haemoglobin content, an additional 2 μl of blood was collected and diluted in 150 μl distilled water. After lysis of erythrocytes and removal of the cell debris, supernatants were transferred into 96-well plates, and optical density at 540 nm was determined using an enzyme-linked immunosorbent assay plate reader. Sampling and measurements were carried out in triplicate for each mouse. A/J and C57BL/6 mice had significantly different levels of haemoglobin (two-way ANOVA, P<0.0001) and maintained this difference throughout the study period. *Indicates statistically significant difference.
Figure 2
Figure 2
Scatter plots illustrating gene expression differences between A/J and C57BL/6 mice. From each strain, groups of mice (n = 5) were killed at days 0, 4, 7, 10 and 17 and liver tissue was snap frozen in liquid nitrogen. Total RNA from liver was extracted and pools of RNA consisting of five individuals/time point/strain were reverse transcribed and labelled with either Cy3 or Cy5 dyes. Differentially labelled cDNAs from each strain were hybridized onto 7000 gene oligo arrays in three replicates/time point. Hybridization signal were analysed using MaxD microarray analysis software. (a) Scatter plot of A/J vs C57BL/6 ratios on days 0 and 4 with every dot representing an individual gene. Among others, genes labelled Apcs, Pbp and abhydrolase domain containing 1 (Abhd1) were higher expressed in A/J mice (ratio >0) as compared to C57BL/6 mice. Vice versa, as examples of genes that were higher expressed in C57BL/6 mice (ratio <0) nicotinamide N-methyltransferase (Nnmt), major urinary protein 4 (Mup4), haemolytic complement (Hc) and Bcl2-interacting killer-like (Biklk) genes are labelled. (b) Scatter plot of A/J vs B57BL/6 ratios on days 4 and 7. Genes such as cytochrome P450, family member 4 (Cyp4a14), carbonic anhydrase (Car3) and catalytic glucose-6-phosphatase (G6pc) are upregulated in A/J, genes labelled adipose differentiation related protein (Adfp), presenilin 2 (Psen2), structural p-lysozyme (Lzp-s) and steaoryl-coenzyme A desaturase 1 (Scd1) are upregulated in C57BL/6 mice. (c) Scatter plot of A/J vs C57BL/6 ratios on days 7 and 10. More higher expressed genes were (among others) interferon alpha/beta receptor 1 (Ifnar1), chemokine (C-X-C motif) ligand 13 (Cxcl 13), chemokine (C-C motif) ligands 8 and 6 in A/J and chemokine (C-X-C motif) ligand 9, interferon-activated gene 203 (Ifi203) and bone marrow stromal cell antigen 1 (Bst1) in C57BL/6. (d) Scatter plot of A/J vs C57BL/6 ratios on days 10 and 17. Serum amyloid A 2 (Saa2), Apo A4, K inward rectifying channel family J 10 (Kcnj10), chemokine (C-C motif) ligand 24 and orosomucoid 1 (Orm1) appeared higher expressed in the A/J, whereas selen binding protein 2(Selenbp2), serin/threonin kinase 25 (Stk25) and glutathione S-transferase alpha 2 (Gsta2) were higher expressed in the C57Bl/6 strain.
Figure 3
Figure 3
Hierarchical cluster of mean gene expression levels. Single-linkage hierarchical cluster analysis of the genes upregulated in A/J (red) or C57BL/6 mice (green) over each of the time points was performed using the MaxD software.
Figure 4
Figure 4
Cluster analysis revealing strain-specific expression patterns. Single-linkage hierarchical cluster analysis of the genes upregulated in A/J (red) or C57BL/6 mice (green) over each of the time points was performed using the MaxD software.
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References

    1. Torr SJ, Hargrove JW, Vale GA. Towards a rational policy for dealing with tsetse. Trends Parasitol. 2005;21:537–541. - PubMed
    1. Mansfield JM, Paulnock DM. Regulation of innate and acquired immunity in African trypanosomiasis. Parasite Immunol. 2005;27:361–371. - PubMed
    1. Omamo SW, d’Ieteren GD. Managing animal trypanosomosis in Africa: issues and options. Rev Sci Tech. 2003;22:989–1002. - PubMed
    1. Nyame AK, Kawar ZS, Cummings RD. Antigenic glycans in parasitic infections: implications for vaccines and diagnostics. Arch Biochem Biophys. 2004;426:182–200. - PubMed
    1. Hill EW, O’Gorman GM, Agaba M, Gibson JP, Hanotte O, Kemp SJ, et al. Understanding bovine trypanosomiasis and trypanotolerance: the promise of functional genomics. Vet Immunol Immunopathol. 2005;105:247–258. - PubMed

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