Elevated Immune Mediators in Lyme Disease.

<p>Serum samples from patients with diagnosed acute Lyme disease (n = 44, red) and healthy controls (n = 23, black) were assayed for the presence of 58 soluble mediators and 7 acute phase proteins using an optimized multiplex-based assay system. Displayed are those mediators that show significant changes (q<0.1%) in Lyme patients vs. controls. (Panel A) Results are displayed as a heat map to visualize differences in mediator levels in Acute Lyme patients relative to controls. (Panel B) Unsupervised hierarchical clustering of the results was performed, and the output displayed as a heatmap. This analysis resulted in the formation of two clusters, including a “mediator high” cluster that contains samples derived from patients with acute <i>B. burgdorferi</i> infection who exhibited elevated serum inflammatory mediators. The second “mediator low” cluster includes a subset of samples from acute <i>B. burgdorferi</i> infection as well as the matched healthy controls, both of which exhibited low levels of inflammatory mediators.</p

Chemokine levels in Lyme disease before and after treatment.

<p>Displayed are the levels of the chemokines CXCL10 (Panel A), CCL19 (Panel B), CXCL9 (Panel C) and CXCL8 (Panel D) measured in the serum of Lyme patients (n = 44) pre-treatment (acute disease) and post-treatment (4 weeks following diagnosis) as compared to healthy controls (n = 23). Horizontal bars represent the medians for each sample group.</p

Serum Amyloid A levels are associated with elevated liver function tests.

<p>Lyme patients were separated into two populations based on normal (n = 24) vs. elevated liver enzyme tests (n = 20) and the levels of Serum Amyloid A (Panel A) and CRP (Panel B) were compared at multiple time points relative to healthy controls (n = 23). Serum amyloid A (p = 0.036) and CRP (p = 0.017) levels are significantly different between Lyme patients with high liver function tests at the pre-treatment visit. Both groups are significantly different from controls at (p<0.0005), but are not different from each other or controls, at both the Post-treatment and 6 Month follow-up visits. There is no significant difference in CRP levels between high and normal liver function groups, however both are significantly different from controls (p<0.0005).</p

Correlation Analysis of Key Mediators.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093243#pone-0093243-t003" target="_blank">Table 3</a> shows the relationship between acute, pre-treatment levels of key immune mediators of early Lyme disease as discussed in this paper. Pearson correlations were used for all analyses. A significant correlation can be seen between CXCL10, CXCL9 and CCL19. CRP shows a significant positive correlation with Serum Amyloid A (SAA). IL-6 does not correlate with either CRP or SAA.</p

Acute Lyme Disease patient subsets defined by circulating mediators versus clinical phenotypes.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093243#pone-0093243-t002" target="_blank">Table 2</a> shows group differences based on the heat map generated using SAM (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093243#pone-0093243-g001" target="_blank">Figure 1</a>). No demographic differences between groups were seen. Categorical variables were compared using Fisher's Exact tests, while continuous variables were compared using unpaired t-tests. Lymphocyte values given are mean (standard deviation), while other values given are a percentage of the respective subset. Number of symptoms pre-treatment is defined as the number of symptoms reported by the patient during structured interview by the principle investigator (JNA) or study staff (LAC). Illness duration is defined as the period of time between first sign or symptom of disease and presentation for treatment and enrollment in the study.</p>a<p>Acute Lyme Mediator High n = 26; Acute Lyme Mediator Low n = 17;</p>b<p>Acute Lyme Mediator High n = 27; Acute Lyme Mediator Low n = 15.</p

Serostatus versus T Cell Chemokine Levels.

<p>Cutoffs in early Lyme disease cases were created for CXCL10, CXCL9, and CCL19 based on being higher or lower than controls. The columns show differences between those who are sero-positive at either time of diagnosis or immediately following treatment and those who are negative at both time points for these three biomarkers. CXCL10 and CXCL9 show differences in the association between those who are sero-positive and sero-negative (p = 0.003 and p = 0.004, respectively). There is no statistical difference in the association between serogroups for CCL19.</p