What is clear from the RT-qPCR result is that IFNG and IL17A are expressed to a greater extent in DBA/2 compared to C57BL/6 mice. The upregulation of

ISG20 in DBA/2 mice Protein Tyrosine Kinase inhibitor originally identified by microarray analysis was also not confirmed by RT-qPCR analysis (Figure 7). The probe set on the microarray (103432_at) and the TaqMan assay (Mm00469585_m1) for ISG20 (NM_001113527) target different regions of this transcript (i.e. 2nd and 3rd versus 1st and 2nd exons, respectively) so alternative splicing could account for the discrepancy [47]. GSK2245840 C. immitis infection also resulted in the downregulation of genes in DBA/2 versus C57BL/6 mice (Figures 2 and 3), which was confirmed by RT-qPCR (Figure 7, S3A and S3B). THBS1 encodes thrombospondin, an extracellular protein that binds a large number of substrates (calcium, heparan sulfate, integrins, the CD36 macrophage scavenger receptor, and transforming growth factor beta 1 [TGF-β])

to modulate cellular attachment, migration, differentiation, and proliferation [48]. IFN-γ appears to regulate THBS1 at the post-transcriptional level in keratinocytes and downregulates THBS1 mRNA in conjunction with TNF-α [28]. THBS1-deficient mice have spontaneous pneumonia that leads to pulmonary hemorrhage, macrophage infiltrations and permanent damage to the lungs, which suggests that this protein is important for maintaining normal pulmonary homeostasis by limiting the extent and/or duration of inflammation [48]. Therefore, it is possible that the downregulation of THBS1 click here at day 16 in DBA/2 mice facilitates inflammatory responses that contribute to resistance to C. immitis infection, but may also contribute to the long term damage to the lung of DBA/2 mice that eventually leads to their death [49]. Downregulation of LYVE1 in DBA/2 versus C57BL/6 mice is also consistent with a stronger inflammatory response in DBA/2 mice following C. immitis infection. Johnson et al.[50] previously demonstrated

that an inflammatory response induced in primary human dermal lymphatic endothelial cells through treatment with TNF-α led to the downregulation of LYVE1 at the transcriptional level. The LYVE1 gene codes for a type I integral membrane receptor that was thought to function in hyaluronan clearance and hyaluronan-mediated leukocyte PI-1840 adhesion, although this biological role has not been confirmed in knockout mice [50, 51]. Consistent with the role of TNF-α in modulating expression of both of these genes (THBS1 and LYVE1) we found that TNF-α was more highly expressed in DBA/2 mice at day 14 by both microarray (fold change of 3.43, data not shown) and RT-qPCR analysis (Figure 7). Protein interaction network analysis identified the transcription factor HIF1A as a network hub. HIF1A was upregulated to a greater extent at day 14 in resistant DBA/2 versus susceptible C57BL/6 mice, and this was confirmed by RT-qPCR (Figure 7).