Background <p>We aimed to investigate whether substances secreted by <i>Clonorchis sinensis</i> excretory/secretory protein (CS-ESP) have an effect on the inflammation of rheumatoid arthritis (RA) and ankylosing spondylitis (AS) and to identify specific peptides through related proteomic analysis to determine which proteins exhibit anti-inflammatory effects more specifically.</p> Methods <p>Peripheral blood mononuclear cells (PBMCs) were obtained from healthy controls (HCs), RA and AS patients. Cytotoxicity of CS-ESP at different doses was assessed by MTS and flow cytometry before performing experiments. Inflammatory cytokine producing cells were analyzed using flow cytometry. To determine the effect of CS-ESP in an arthritis mouse model, 8-week-old SKG mice were injected intraperitoneally with curdlan and treated with CS-ESP; body weight and paw swelling were checked twice a week. Inflammation was evaluated using immunohistochemistry. We conducted proteomic analysis on CS-ESP and identified specific Cs-GT and Cs-Severin proteins. In vitro effect of coculture with Cs-GT and Cs-Severin was determined by inflammatory cytokine measurements.</p> Result <p>Treatment with CS-ESP resulted in no reduced cell viability of PBMCs. In experiments culturing PBMCs, the frequencies of IL-17A and GM-CSF producing cells were significantly reduced after CS-ESP treatment. In the SKG mouse model, CS-ESP treatment significantly suppressed clinical score, arthritis and enthesitis. Treatment with Cs-GT and Cs-Severin resulted in no reduced cell viability of HC PBMCs. After Cs-GT and Cs-Severin treatment of HC PBMC, the frequencies of IL-17A and GM-CSF producing cells were significantly reduced.</p> Conclusions <p>We provide evidence showing that CS-ESP, Cs-GT and Cs-Severin can ameliorate clinical signs and cytokine derangements in AS.</p> Graphical Abstract <p></p>