Table 4.
Quantification of leptospires and cytokine expression in the kidneys of infected mice. We employed a digital PCR approach specifically targeting the lipl32 gene (for detecting pathogenic Leptospira spp.) and the fliG gene (for differential detection of pathogenic and nonpathogenic Leptospira spp.) to assess the number of leptospires in kidneys. To evaluate the effect of the Leptospira presence on cytokine expression in infected mice, we quantified the expression levels of cytokines, IL-10, IL-6 and TNF-α, using a real-time qPCR assay.
| After infection at day 7 |
Without infection |
Nonpathogenic L. biflexa infection |
Pathogenic L. interrogans infection |
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|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| #1 | #2 | #3 | #1 | #2 | #3 | #4 | #1 | #2 | #3 | #4 | #5 | #6 | #7 | #8 | #9 | #10 | #11 | #12 | ||
| lipl32 | Concentration (copies/ μg RNA) |
0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 6.3 | 15.2 | 0.0 | 0.0 | 0.0 | 4.2 | 6.6 | 34.4 | 0.0 | 6.7 | 9.8 | 0.0 |
| fliG | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 2.1 | 8.4 | 2.2 | 8.8 | 4.4 | 4.4 | 6.6 | 8.7 | 0.0 | 12.9 | 0.0 | 6.5 | 4.4 | |
| IL-10a | Expression level (Fold change) |
1.0 | 1.0 | 1.0 | 1.4 | 1.0 | 2.9 | 3.0 | 13.9 | 17.8 | 7.6 | 12.2 | 12.6 | 28.2 | 18.7 | 15.8 | 189.0 | 29.9 | 13.3 | 11.8 |
| IL-6a | 1.0 | 1.0 | 1.0 | 0.4 | 0.9 | 1.2 | 1.0 | 4.4 | 3.2 | 2.4 | 2.0 | 2.1 | 4.7 | 9.8 | 4.9 | 2.3 | 9.7 | 3.1 | 2.8 | |
| TNF-αa | 1.0 | 1.0 | 1.0 | 1.0 | 1.4 | 1.5 | 1.1 | 2.6 | 1.7 | 1.4 | 1.7 | 2.1 | 3.1 | 2.7 | 2.0 | 2.8 | 2.9 | 1.9 | 1.1 | |
| After infection at day 28 |
Without infection |
Nonpathogenic L. biflexa infection |
Pathogenic L. interrogans infection |
|||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| #1 | #2 | #3 | #1 | #2 | #3 | #4 | #1 | #2 | #3 | #4 | #5 | #6 | #7 | #8 | #9 | #10 | #11 | #12 | ||
| lipl32 | Concentration (copies/ μg RNA) |
0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 1680.4 | 77.8 | 135.1 | 597.2 | 49.3 | 13.3 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 285.8 |
| fliG | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 4.2 | 2.2 | 2.2 | 2.2 | 4.4 | 4.4 | 2.2 | 2.1 | 0.0 | 0.0 | 2.2 | 6.5 | |
| IL-10a | Expression level (Fold change) |
1.0 | 1.0 | 1.0 | 6.9 | 5.7 | 12.8 | 9.3 | 2.9 | 0.6 | 3.4 | 2.5 | 5.0 | 5.2 | 3.8 | 2.5 | 2.1 | 0.6 | 4.8 | 1.8 |
| IL-6a | 1.0 | 1.0 | 1.0 | 2.5 | 2.2 | 2.8 | 2.9 | 1.7 | 1.1 | 2.3 | 1.9 | 3.4 | 1.8 | 1.5 | 1.5 | 1.0 | 0.9 | 1.7 | 2.0 | |
| TNF-αa | 1.0 | 1.0 | 1.0 | 5.0 | 3.7 | 5.0 | 4.8 | 1.8 | 1.2 | 2.3 | 1.8 | 4.2 | 2.5 | 1.9 | 1.7 | 1.2 | 1.6 | 2.7 | 1.4 | |
a
TaqMan probes were used for the indicated genes: Mm00439614_m1 (interleukin (IL)−10), Mm01210733_m1 (IL-6), Mm99999068_m1 (TNF-α) and Mm99999915_g1 (GAPDH).