The detection limits for each hormone measured FDA-approved Drug Library were the following: cortisol, 0.4 μg/dL; DHEA-S, 2 μg/dL; estradiol, 2 pg/mL; prolactin, 0.25 ng/mL and testosterone, 0.1 ng/dL. IFN-γ, IL-10 and TNF-α (Pharmingen, San Diego, CA) levels were measured in cell culture supernatants using commercially available ELISA kits. Culture supernatants were harvested at 96 h for IFN-γ, 48 h for IL-10 and 24 h for TNF-α. Assays were performed according to the manufacturer’s instructions. The detection limits for each cytokine were as follows: IFN-γ, 55 pg/mL; IL-10, 3 pg/mL and TNF-α, 3 pg/mL. Hormone concentrations in controls
and patients were compared using the Mann–Whitney test. Correlations between the levels of cytokines and hormones were evaluated with the
Spearman test. All statistical tests were performed using GraphPad software version 5.0 (GraphPad Software Inc., San Diego, CA, USA). To determine whether hormone levels were associated with LCL, we assessed plasma levels of cortisol, estradiol, DHEA-S, prolactin and testosterone in NV and LCL patients with an active cutaneous lesion. The clinical profile of the patients analyzed (n = 57) is shown in Table 1. LCL patients (n = 32) had lower plasma levels of DHEA-S, prolactin and testosterone than NV (n = 32; Fig. 1C, D and F). No difference between patients and controls was AZD8055 cell line observed
in levels of cortisol or estradiol ( Fig. 1A and B). Possible correlations between hormone levels and clinical or immunological parameters, such as lesion size, healing time, Glucantime dosage and SLA-stimulated IFN-γ, IL-10 and TNF-α levels were analyzed using the Spearman test. We tested the correlation between each hormone and each clinical parameter or cytokine. Cortisol showed a positive correlation with healing time and dose of Glucantime used in the treatment and a negative correlation with in vitro SLA-stimulated IFN-γ levels (Fig. 2A–C). For estradiol, males were analyzed separately from females because of the considerable difference in the concentration Selleckchem Osimertinib of this hormone between the two groups. Plasma levels of estradiol in males correlated positively with lesion size, whereas in females, a correlation was observed with total dose of Glucantime used in treatment (Fig. 3A and B). Prolactin correlated positively with lesion size and negatively with in vitro IFN-γ levels (Fig. 4A and B). Other correlations tested did not reach statistical significance. To evaluate whether hormone level changes were similar in males and females with LCL, each group was tested separately. Male patients (n = 17) showed a reduction in levels of DHEA-S, prolactin and testosterone compared with controls ( Fig. 5 and Fig. 6C and E).