In an effort to better understand factors that may play a role in the development of Gulf War Illness, we examine the effects of 3 organic compounds and 9 metals of interest to the military population that served during the Gulf War on the function and stability of a model of the human blood brain barrier (BBB). Using a BBB model consisting of human brain endothelial cell and astrocyte cell co-cultures in a transwell membrane system, we exposed transwells to organic compounds and metals either individually or as a combination of an organic followed by a metal and assessed changes in BBB function. Assays included trans-epithelial electrical resistance (TEER), translocation of two different sized fluorescence markers or metals of interest from the luminal side to the abluminal side of the BBB, and changes in expression of tight-junction proteins and cytokines. Several co-exposures resulted in reduced TEER values, although minimal or no effects were seen on transfer of the fluorescent markers and metals across the membrane. Expression of tight junction proteins was also unaffected, but some cytokines increased synergistically with several co-exposures. Of all combinations, pyridostigmine bromide, permethrin, and depleted uranium had the most consistent effects. The cause of Gulf War Illness is not well understood, and interactions of pesticides, prophylactics, and metals may factor into symptom development more than any single exposure. Understanding these interactions may also shed light on risk factors for other populations potentially exposed to similar contaminants in their environment.