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The aim of this study was to examine the effect of the level-1 Yo-Yo intermittent recovery test (YYIRT1) on polarization of macrophages in young football players.Fourteen male football players (19.9 ± 1.4 years old) were enrolled in this study. YYIRT1 was performed with 20-meter shuttle runs at increasing speeds and 10-second active recovery in a 5-meter distance between runs till exhaustion. Fasting blood samples were collected before and immediately after YYIRT1. Analysis for macrophage polarization by flow cytometry, reactive oxygen species (ROS) by flow cytometry, biochemical parameters by chemical reactions, and serum cytokines by ELISA were performed. The rating of perceived exertion (RPE) and cardiovascular parameters were recorded.The time to exhaustion was 714.1 ± 114.4 seconds. The oxygen uptake () was 48.7 ± 5.6 mL/min/kg, RPE scale was 19 ± 1, resting heart rate and maximal heart rate were 64.9 ± 8.8 beat/min and 181.9 ± 9.3 beat/min, respectively, indicating a high level of cardiopulmonary fitness. The expression of macrophage-specific CD14 and M1 marker HLA-ABC, but not M2 marker CD206, was down-regulated after YYIRT1. The intracellular ROS levels in macrophages had no significant change. In biochemical profile, the serum levels of lactic dehydrogenase (LDH), a marker of muscle damage, increased after YYIRT1 whereas no significant alteration was noted in creatine phosphokinase (CPK), blood urine nitrogen, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), and C-reactive protein. The serum levels of interleukin (IL)-6, IL-10 and tumor necrosis factor (TNF)-α had no significant change.The YYIRT1 may induce muscle damage accompanied by modulation of macrophage polarization toward suppression of M1 phenotype in young football players.