Prior studies with men who have sex with men have documented a strong association between substance use with sex and risk for acquisition of HIV. However, few studies have been conducted about gay male couples’ use of substances with sex, despite the fact that between one third and two thirds of men who have sex with men acquire HIV from their relationship partners. The present study sought to (1) describe whether one or both partners in the male couple uses substances with sex—by substance type—within and/or outside of their relationship, and (2) assess whether differences exist in those who use substances with sex within and outside the relationship by the couples’ HIV status. Dyadic data for this analysis were collected in the United States from a nation-wide cross-sectional Internet study about male couples’ relationships and behaviors. Couple-level descriptive and comparative analyses were employed with 361 male couples. Except for alcohol, most couples did not use substances with sex. Of those who did, rates of who used it with sex and substance type within the relationship varied; most couples only had one partner who used substances with sex outside the relationship. Significantly higher proportions of concordantly HIV-negative and HIV-positive couples had both partners who used substances (all types) with sex within their relationship over discordant couples. Most couples had one partner who used outside the relationship; only marijuana and erectile dysfunction medication use with sex significantly differed by couples’ HIV status. Findings indicate the need to conduct additional research for prevention development.