Table 1.
Time table of follow-up visits of AE patients.
Vx = follow-up visit, N = number of cases that adhered to follow-up visit, ΔtVx-tV0 = difference between baseline and follow-up visit.
Table 2.
Level of IgG IHA over time and difference of respective levels compared to baseline.
Vx = follow-up visit with V0 = baseline, N = number, M = mean, SD = standard deviation; MD = median, P25/P75 = 25th & 75th percentile, Min. = minimum, Max. = maximum, Z = Wilcoxon’s Z-Value, p-value = significance level with * considered significant.
Table 3.
Number and percentage of AE patients with detectable Em2+ antibodies at different visits and significance level in binominal test.
Vx = follow-up visit with V0 = baseline, N(%) = total number (percentage of patients) with detectable Em2+, p-value = significance level with * considered significant.
Table 4.
Reactivity (indicated as optical densities OD) of IgG4 to larval extract of E. multilocularis (EmAg) over time and difference of respective levels compared to baseline.
Vx = follow-up visit with V0 = baseline, N = number, M = mean, MD = median, SD = standard deviation; Min. = minimum, Max. = maximum, t = difference in units of standard error, df = degrees of freedom, p-value = significance level with * considered significant.
Table 5.
Levels of total and E.multilocularis-specific IgE in kU/l over time and difference of respective levels compared to baseline.
Vx = follow-up visit with V0 = baseline, N = number, M = mean, MD = median, SD = standard deviation; Min. = minimum, Max. = maximum, t = difference in units of standard error, df = degrees of freedom, p-value = significance level with * considered significant.
Table 6.
Levels of cytokines and chemokines over time.
Cytokines IL-8(CXCL8), IL-9, IL-10 and chemokines MCP-4(CCL13), TARC(CCL17), PARC(CCL18) and LARC(CCL20) were quantified in pg/ml in AE patients over time. Vx = follow-up visit with V0 = baseline, N = number, M = mean, SD = standard deviation; Min. = minimum, Max. = maximum.
Table 7.
Alveolar echinococcus (AE) patients staged into groups with progressive, with stable and cured disease.
The mean age with minimum and maximum ist indicated in brackets. The average age was 58.7 years. The two youngest patients were 19, the oldest patient 106 years old. One third of the patients were between 61 and 75 years old at the time of blood collection, and 51% percent of the patients were female. The youngest age group (0-30years) was the smallest with 8%, and the higher age groups of 31-45y, 46-60y, 61-75y and 76+y represented 16%, 24%, 36% and 16% of all AE patients, respectively.
Fig 1.
The E. multilocularis antigen-specific IgG reactivity (OD) in alveolar echinococcosis (AE) patients and infection-free controls.
The AE patients were staged into those with progressive AE, with stable (<60y), stable (≥60) and cured AE. For the determination of age dependent EmAg specific IgG4 responses, the patients were grouped in five age groups. Blood samples from AE patients were collected at three time points in 2005 (year 1) in 2010 (year 5) and in 2016 (year10). Controls were echinococcosis infection-free healthy blood donors. In (A) the EmAg specific IgG1, in (B) the EmAg specific IgG3 and in (C) the EmAg specific IgG4 responses are shown as mean optical densities (OD) with the 95% confidence intervals for the means (diamonds). The data presented in box plots show the median OD per group with the 25% and 75% quartiles and the 1,5x of the interquartile range with outlier as individual points. In (D) the IgG4 reactivity in AE patients to EmAg is shown as mean OD per age group with the 95% confidence intervals.
Table 8.
Serum concentrations of cytokines and chemokines.
Cytokines IL-8, IL-9 and IL-10 and chemokines MCP-4(CCL13), TARC(CCL17), PARC(CCL-18), LARC(CCL20) concentrations in pg/ml (mean [min, max]) were quantified by specific sandwich ELISA. The alveolar echinococcosis (AE) patients were staged in 4 groups with progressive, stable with equal or younger than 60 years (≤60y), stable older than 60 years (>60), and patients with cured AE. Control individuals were E. multilocularis infection-free. For IL-8: *Tukey-Kramer Test: p = 0.04 Cured vs. Control and vs. Stable and vs. Vital/Progressive; For LARC(CCL20) **Tukey-Kramer Test: p = 0.001 for Cured vs. Control.
Fig 2.
The E. multilocularis antigen inducible cellular production by PBMC of chemokines and cytokines.
In (A) MCP-1(CCL2), MCP-3(CCL7), MCP-4(CCL13), in (B) cytokines IL-10 and IFN-γ and in (C) chemokines CCL17(TARC) and CCL18(PARC) were investigated in patients with progressive, stable, and cured AE, and in E. multilocularis infection-free controls. Freshly isolated and in vitro cultured PBMC (2.5x106/ml) were stimulated with E. multilocularis antigen (EmAg, 12 μg/ml) or remained without stimulation (baseline) for 48 hours. Cytokines and chemokines secreted into cell culture supernatants were quantified with specific ELISA. The concentrations are indicated as mean (net) amounts in pg/ml (with the 5% upper and 95% lower confidence interval) of cytokine or chemokine released from stimulated PBMC minus the cellular production of unstimulated (baseline) PBMC. MCP-1(CCL2), MCP-3(CCL7) and MCP-4(CCL13) production by PBMC was quantified in AE patients with progressive (n = 7) stable (n = 29) and cured disease (n = 10) disease. For IL-10, IFN-γ and PARC(CCL18) and TARC(CCL17) production was investigated in AE patients with progressive (n = 19), stable (n = 27) and cured disease (n = 14), and in E. multilocularis infection-free controls (n = 6).** p<0.01 versus cured or control.
Table 9.
Paired Correlations of serum levels of cytokines and chemokines (in pg/ml) in AE patients, with age (in years), EmAg-specific IgG4 responses (OD values), stage of AE disease (4:progressive, 3:stable, 2:cured; 1:control) (study groups: n = 217 AE patients, n = 30 infection-free controls) and year of sample collection (year 1, 5, 10) are shown.
Table 10.
Pearson’s correlations of cytokine and chemokine levels observed in AE patients are shown.
r = Pearson’s r, p = significance level with * considered significant.
Table 11.
Logistic regression models with variables associated with progressive or cured AE clinical status.
OR = Odds, CI 95% = 95% confidence interval, p = significance level with * considered significant.