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<h4>Background</h4>In recent years, researchers have illuminated many non-skeletal actions of vitamin D including host defense against various pathogens and vaccine immunology. The purpose of our study was to explore the potential association between serum vitamin D levels and measles antibody titers.<h4>Methods</h4>The biochemical profiles and de-identified information were accessed from the 2001 to 2004 National Health and Nutrition Examination Survey (NHANES). Participants were divided into quartiles according to their measles antibody titers.<h4>Results</h4>A total of 5,681 participants were analyzed in our study. Participants in the highest quartile of measles antibody titer had significantly lower serological levels of 25-hydroxyvitamin D [25(OH)D] than those in the lower quartiles (53.90 vs. 58.70 nmol/L, a decrease of 8.18%) (p < 0.001). After full adjustment of confounders, the adjusted ß coefficient of 25(OH)D was -0.006 (p<0.001). A decreasing tendency of 25(OH)D among quartiles of measles antibody titers was obvious (p for trend <0.001). The negative association in seropositive subjects remained statistically significant only in non-Hispanic black population before adjustment for age, gender, and other covariates (p<0.05).<h4>Conclusion</h4>Our study highlights the negative association between serum 25(OH)D levels and measles antibody titers.