To report the expression of macrophage migration inhibitory factor (MIF) in murine lacrimal glands. Six lacrimal glands from 3 female 8-week-old CBA/J mice were analyzed using a gene microarray method (Oligo GEArray mouse inflammatory cytokines and receptors, Catalog No. OMM-011, SuperArray Bioscience) and immunofluorescent staining for MIF. Nine lacrimal glands from 8-week-old CBA/J mice, 9 lacrimal glands from 8-week-old C57BL/6 mice, and 7 lacrimal glands from 8-week-old BALB/c mice were analyzed using real-time reverse transcription-polymerase chain reaction. Five spleens, 5 livers, and 5 serum samples from 5 female 8-week-old CBA/J mice were analyzed using enzyme-linked immunosorbent assay. Microarray analysis revealed that MIF is highly expressed in the murine lacrimal gland. Lacrimal acinar cells stain strongly with anti-MIF antibodies. Real-time reverse transcription-polymerase chain reaction revealed that the MIF mRNA expression level is lower in lacrimal glands of CBA/J mice compared with C57BL/6 and BALB/c mice when normalized against the expression of beta-actin mRNA. Enzyme-linked immunosorbent assay revealed that MIF level was higher in lacrimal glands and spleens compared with livers and sera. The murine lacrimal gland expresses high levels of macrophage MIF without any evidence of lacrimal gland inflammation.