While cannabis use is common among people with HIV (PWH), there have been few studies examining the association of use with health outcomes among PWH. We aimed to evaluate the association between cannabis use and bothersome physical and mental health symptoms using both self-report and a direct biomarker for cannabis use. The Medications, Alcohol and Substance use in HIV Study (MASH) is a cross-cohort study focused on polypharmacy and substance use among PWH. Participants were enrolled from October 2018 to May 2022 in the Swiss HIV Cohort Study (SHCS), Kaiser Permanente Northern California (KPNC) HIV Cohort, and 7 sites (Atlanta, GA; Bronx, NY; Washington, DC; Houston, TX; Los Angeles, CA; New York, NY; and Nashville, TN) in the Veterans Aging Cohort Study - HIV (VACS-HIV). Participants submitted self-reported information on the presence/absence of symptoms in the prior 4 weeks, along with bothersome level, using the HIV Symptom Index, which is comprised of 20 symptoms commonly reported in PWH, including fatigue, diarrhea, fever, anxiety, and weight loss, among others. Concurrent self-reported substance use information was also collected, and participants provided nail clippings to be tested for cannabis use. We used logistic regression models, adjusted for age, sex, race/ethnicity, and cohort, to examine associations of self-report and biomarker-identified cannabis use with self-reported bothersome symptoms. The analytic sample included 1,226 PWH who were predominantly male (85%), non-white (64%), with an average age of 57 years; 20% tested positive for cannabis use, and 23% reported recent cannabis use. Agreement between self-report and biomarker cannabis use was substantial (agreement 89%, kappa = 0.67). Put together, 27% either reported cannabis use or tested positive. Of the 20 symptoms included in the HIV Symptom Index, 19 were more prevalent at a bothersome level among those with cannabis use compared to those without. In adjusted models, the patterns of association remained for both self-reported and biomarker-confirmed cannabis use, and they were particularly substantial for memory, depression, anxiety, and nausea. Cannabis use was common among PWH and was associated with a number of bothersome symptoms. While cannabis use may be used to treat loss of appetite, nausea/vomiting, and weight loss, the associations of cannabis use with depression, anxiety, and memory loss are concerning and deserve investigation.