This study aimed to document the vitamin D status of HIV-infected individuals across a wide latitude range in one country and to examine associated risk factors for low vitamin D. Using data from patients attending four HIV specialist clinics across a wide latitude range in Australia, we constructed logistic regression models to investigate risk factors associated with 25(OH)D<75 nmol/L. 1788 patients were included; 87% were male, 76% Caucasian and 72% on antiretroviral therapy. The proportion with 25(OH)D<50 nmol/L was 27%, and <75 nmol/L was 54%. Living in Melbourne compared to Cairns (adjusted odds ratio (aOR) 3.30; 95%CI 2.18, 4.99, p<0.001) and non-Caucasian origin (aOR 2.82, 95%CI 2.12, 3.75, p<0.001) was associated with an increased risk, while extreme UV index compared to low UV index was associated with a reduced risk (aOR 0.33; 95%CI 0.20, 0.55, p<0.001) of 25(OH)D <75 nmol/L. In those with biochemistry available (n=1117), antiretroviral therapy was associated with 25(OH)D <75 nmol/L, however, this association was modified by serum cholesterol status. Location and UV index were the strongest factors associated with 25(OH)D <75 nmol/L. Cholesterol, the product of an alternative steroid pathway with a common precursor steroid, modified the effect of antiretroviral therapy on serum 25(OH)D. This article is protected by copyright. All rights reserved.