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Though many alternative methods to tuberculin skin test (TST) have been established and evaluated in recent years, sensitivities and specificities of most methods could not meet the requirements of golden standards.In this study, we sought to identify whether repeated TSTs could affect the immune responses in experimental monkeys.Nine natural tuberculosis (TB) monkeys receiving repeated TSTs biweekly were used to demonstrate the effect on TST responsiveness.Two healthy monkeys were administrated with repeated TSTs to analyze the immune response profiling.Intrapalpebral reactions in TB infections gradually weakened or presented intermittent positive reactions.