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Progressive multifocal leukoencephalopathy (PML) is an opportunistic infection due to reactivation of John Cunningham virus (JCV). The diagnosis depends on evidence from clinical, imaging, and virologic studies. When the cerebrospinal fluid shows a negative polymerase chain reaction result, brain biopsy is required to confirm the diagnosis. PML has no standard treatment except for immune reconstitution. The anti-JCV effect of mefloquine, however, is supported by some studies, and if brain biopsy is difficult, a mefloquine trial can be considered. We describe a case of possible PML successfully treated with mefloquine.