The clinical spectrum of cutaneous leishmaniasis (CL), an intracellular parasitic pathogen, ranges from a single sore healing to chronic crusty lesions with a manifestation of treatment resistance. The complicated interaction between Leishmania bodies and the early immune response, including innate and adaptive mechanisms, determines the evolution of nodules. This study examined the levels of the chemoattractant interleukin 8 (IL-8), pro-inflammatory nitric oxide (NO), and immunoregulatory macrophage inhibitory factor (MIF) in the serum of subjects recently diagnosed with cutaneous leishmaniasis, in parallel with patients being monitored during consecutive sodium stibogluconate (Pentostam) treatment. A total of 161 serum samples of newly diagnosed individuals and patients undergoing pentostam injections were collected form an endemic area of Diyala, east central of Iraq. Sandwich ELISA was used to measure the level of IL-8, NO and MIF in the studied groups. Results of circulatory markers levels showed a considerable difference in all groups, with IL-8 being exceptionally higher in the first two groups of pretreated and dose-1 (191.5, 273.64) pg/ml respectively, while NO was found to be lower than in control subjects, particularly in the pretreated group (12.08 μmol/L) and MIF level was significantly higher in the pretreated group, which was (7.18 pg/ml). These findings can provide insights for distinction of disease phase and monitoring treatment efficacy along consecutive dosages, particularly in populations where CL is endemic.