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Background: To evaluate the influence of previous physical activity (PA) during childhood, adolescence, and current PA practice on the production of antibodies and inflammatory response between the first and second doses of the COVID-19 vaccine. Methods: Fifty-nine men and 56 women were evaluated before the first vaccine, and 12 weeks later, blood samples were taken to quantify production of anti-severe acute respiratory syndrome coronavirus-2 immunoglobulin G antibodies and cytokines. Previous PA during childhood and adolescence was self-referred, and current PA was assessed using the International Physical Activity Questionnaire. Results: A positive and significant association was observed only between PA practice during adolescence and an increase in antibody production in adulthood (β = 2012.077, 95% confidence interval, 257.7953–3766.358, P =.025). Individuals who practiced PA during adolescence showed higher production of antibodies between the first and second vaccine dose compared to nonpractitioners (P =.025) and those that accumulated ≥150 minutes per week of current moderate–vigorous PA (MVPA), and presented higher antibody production in relation to who did <150 minutes per week of MVPA (P =.046). Individuals that were practitioners during childhood produced higher G-CSF (P =.047), and those that accumulated ≥150 minutes per week of current MVPA demonstrated lower IP-10 levels (P =.033). However, PA practitioners during adolescence presented higher G-CSF (P =.025), IL-17 (P =.038), IL-1RA (P =.005), IL-1β (P =.020), and IL-2 (P =.026) levels. Conclusion: Our results suggest that adults that accumulated at least 150 minutes of MVPA per week or practiced PA during adolescence developed an improved immune and inflammatory response against COVID-19 vaccination. [ABSTRACT FROM AUTHOR]