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Previous research has indicated that genetic and environmental factors shape physiological activity. Cortisol levels, in particular, have received significant attention, with studies indicating substantive heritability estimates across various sampling techniques. A related line of research has indicated that genetic and environmental factors that explain variability in cortisol levels may vary across context and experiences by way of gene-environment interactions (G × Es). Despite these findings, a limited number of studies have examined the extent to which interpersonal relationships may operate as a moderator. The current study focused on co-twin relationship quality as a source of moderation, as twins are more likely to have contact with one another and to form close, interpersonal relationships with their co-twin relative to singleton siblings. Using a sample of 298 adult twins from the National Survey of Midlife Development in the United States (MIDUS), we examined the extent to which genetic and environmental factors that explain variability in urinary cortisol levels varied across levels of co-twin relationship quality. The heritability of cortisol levels was greater and nonshared environmental influences were lower at greater levels of relationship quality. These findings suggest that the heritability of cortisol may vary across context, and positive relationships with others may moderate such factors., (Copyright © 2019 Elsevier Ltd. All rights reserved.)