Background: Cytokines contribute to insulin resistance in pregnancy, but the role of distinct cytokines is not fully understood. Objectives: To study whether cytokines produced by tissues other than skeletal muscle are associated with glucose and insulin metabolism activity in overweight and obese women and to study whether these associations can be modified by physical activity. Methods: A longitudinal study with 44 overweight and obese pregnant women was conducted. Changes in cytokines levels (IFN-gamma, IP-10, IL1-alpha MIP1-alpha, adiponectin and leptin) and ICAM1 from early (15 wk) to late (32 wk) pregnancy were determined. Physical activity was measured objectively with accelerometers. In linear regression models, the associations between (changes in) cytokine levels and fasting glucose, fasting insulin and HOMA-IR were studied. Results: Both IFN-gamma and IP-10 levels increased from early to late pregnancy, and adiponectin levels decreased. IFN-gamma and IP-10 were positively associated with fasting glucose, whereas IL-1 alpha , ICAM1 and adiponectin were inversely associated with insulin and insulin resistance. The association of IL-l alpha with insulin and insulin resistance was only found in women with low levels of physical activity. Conclusions: IFN-gamma, IP-10, IL1-alpha, ICAM1, and adiponectin may play a role in glucose and insulin metabolism in pregnancy. The relationship of IL-1 alpha with insulin and insulin resistance might be moderated by levels of physical activity. Further studies are required to confirm the role of these cytokines in glucose and insulin metabolism in obese pregnant women. (C) 2015 Elsevier Ltd. All rights reserved.