Tolouei et al. : IL-23 and IL-27 Levels in Macrophages Collected … acts on the bound enzyme was added. The absorbance of each well was read at 450nm. The concentration of IL-23 and IL-27 in the treated and untreated supernatants was deter- mined using standard curve. Cytokine values were expressed as picogram/ml (pg/ml). Statistical analysis Statistical analysis was performed using SPSS version 16. Student's paired and inde- pendent t test were used to determine wheth- er significant difference exists between the cytokine levels. Results Among 20 patients recruited with proven CL, 18 were male and 2 were female. The basic information of the study is summarized in Table 1. Molecular analysis All samples were undertaken microscopic examinations and PCR assay. PCR experi- ment revealed a single 625bp DNA fragment was seen on the gel which is accordance with DNA fragment of L. major reference (Fig. 1). Cytokine production IL-23 levels The level of IL-23 in healing and non-healing form of CL before stimulation was 182.9±23.9 pg/ml and 141.3±13.5 pg/ml respectively which was significantly different (P < 0.001). The level of IL-23 after stimulation in super- natants of macrophages in healing and non- healing form of CL was 386 ±43.9 pg/ml vs. 235 ±43.9 pg/ml which was significantly dif- ferent (P < 0.001). The level of IL-23 before and after stimulation in supernatants of mac- rophages in healing form of CL was 182.9±23.9pg/ml and 386.9±43.9 pg/ml, re- spectively, whereas the level of IL-23 in non- healing form was 141.3±13.5pg/ml and 235.5±43.9pg/ml, respectively. The differ- ences between IL-23 level in supernatant of the macrophage culture before and after stimu- lation in patients with healing and non-healing form of CL were 203.9±38.3 vs. 94.1±36.5 respectively .The data were shown the differ- ence in healing form of CL was significantly higher than non-healing form of CL (P < 0.001). IL-27 level The concentration of IL-27 in healing and non- healing form of CL before stimulation was 36.6±4.5 pg/ml and 25.4±4.2 pg/ml respec- tively which was significantly different (P < 0.001). The level of IL-27 after stimulation in supernatants of macrophages in healing and non-healing form of CL was 73.3 ±6.8 pg/ml vs. 46.2±9.2 pg/ml which was significantly different (P < 0.001). The concentration of IL- 27 before and after stimulation in supernatants of macrophages in healing form of CL was 36.6±4.5pg/ml and 73.3±6.8pg/ml, respec- tively, whereas the level of IL-27 in non-heal- ing form was 25.4±4.2pg/ml vs. 46.2±9.2pg/ml. There was a significant difference (P < 0.001) in IL-27 level before and after stimulation in healing and non-healing form of CL. The differences between IL-27 concentration in supernatant of the macrophage culture be- fore and after stimulation in patients with healing form and non-healing form of CL was 36.7±6.48pg/ml vs. 20.8±7.78pg/ml re- spectively. The data were shown the differ- ence of IL-27 in healing form of CL was sig- nificantly higher than non- healing form of CL (P < 0.001). Available at: http://ijpa.tums.ac.ir 21