Immune laboratory methods
Flow cytometry was used to analyze the CD4+ Th cell population in whole
blood from the patients at randomization, and one and three years after
completed ILIT. Peripheral blood mononuclear cells obtained from the
patients at randomization and one year after ILIT were stimulatedin vitro with birch and timothy allergen. Levels of IL-5, IL-10,
IL-13, IFN-γ, CCL17 and CXCL10 were quantified using Luminex. For
detailed methods, experimental protocols, and statistical analyses, see
the Methods section in this article’s supplementary information.
Statistics
Descriptive statistics for RQLQ, RTSS and MS are presented in medians
and percentiles (p25 and p75), and in the graphs with medians and 95%
confidence interval. Paired comparisons over time were calculated with
Friedman’s test and adjusted with the Bonferroni correction for multiple
comparisons. Descriptive statistics for IgE, IgG4, SPT and CAPT are
presented in mean values and standard deviation (SD). Paired comparisons
over time were calculated with repeated measures ANOVA with Bonferroni
confidence interval adjustment. The answers to the 28 questions in RQLQ
were explored with an item analysis, rendering a Cronbach’s Alpha at
0.933; thus the changes within the different domains of RQLQ were
consistent. All analyses above were performed in SPSS version 25 (IBM
Corp. Armonk, NY, USA).
All flow cytometry, cytokine and chemokine data were analyzed using
GraphPad Prism, version 8.3.1 (GraphPad software, Inc., La Jolla, CA,
USA), and non-parametric tests were used. Comparisons at the different
time points within the treatment groups were calculated using paired
Wilcoxon signed ranks test. Unpaired Mann Whitney U test was used
to compare differences between the treatment groups at the different
time points. The significance level was set at p < 0.05.