In 1981 and 1982, two US citizens died from Japanese encephalitis (JE) acquired in China. In 1983, the Centers for Disease Control initiated an evaluation of a purified, inactivated, mouse-brain-derived JE vaccine produced and used in Japan since 1966. Two doses of this vaccine given 1–2 weeks apart evoked neutralizing antibody titers ≥8 in only 77% of recipients. After three JE vaccine doses administered 1–2 weeks apart, 99% developed titers ≥8. When a third dose was given to 29 participants 6–12 months after the primary series, all developed titers ≥16. Reported adverse reactions included injection site tenderness (18%), erythema (6%), or swelling (3%); headache (9%); and dizziness, fatigue, sleepiness, nausea, chills, fever, or lower back pain (≤5%). On the basis of this study, three doses of BIKEN JE vaccine are recommended for US citizens who may be at risk of exposure to JE virus.