Objectives: High-intensity exercise induces intensity dependent leukocytosis due to increased trafficking of white blood cells in circulation and also causes red blood cell (RBC) damage and hemolysis due to osmotic and mechanical stress. The present study was aimed to investigate the high-intensity exercise-induced perturbations of hematological profile in sedentary post-pubertal boys and girls. Materials and Methods: Blood samples were collected from sedentary post-pubertal boys (n = 22, age = 16.10 ± 0.74 years) and girls (n = 22, age = 16.04 ± 0.63 years) before and immediately after exercise to assess hematological parameters such as RBC Count, hematocrit, hemoglobin concentration (Hb), total leukocyte counts, and differential count. Results: Pre-and post-exercise RBC count, Hb and hematocrit had no significant inter-group variation. Pre-and post-exercise hematocrit and Hb were significantly (P < 0.001) higher in post-pubertal boys. Leukocyte count had insignificant intergroup variation before the exercise but it increased significantly (P < 0.001) following exercise in both the groups. Monocytosis and neutrophilia were significantly (P < 0.01) higher in post-pubertal boys. Percentage change in monocyte, eosinophil and basophil did not depict significant inter-group variation while percentage increase in neutrophil was significantly (P < 0.01) higher in boys as compared to girls. Post-exercise absolute lymphocyte count as well as percentage increase in this variable was significantly (P < 0.001) higher in girls as compared to boys. Absolute eosinophil count increased significantly in both the groups but its relative count declined substantially probably due to higher rate of mobilization of lymphocyte and neutrophil. Basophil count was also perturbed following exercise. Conclusion: Gender appeared to have insignificant impact on exercise-induced perturbation in hematological profile at post-pubertal stage except for neutrophil and lymphocyte. ©2020 Published by Scientific Scholar on behalf of Indian Journal of Physiology and Pharmacology.