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A Chemokine, Interferon (IFN)--Inducible Protein 10 kDa, Is Stimulated by IFN- and Recruits Immune Cells in the Ovine Endometrium¹

^a Laboratory of Animal Breeding, Faculty of Agriculture, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan ^b National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan ^c Reproduction Research Unit, USDA-ARS, U.S. Meat Animal Research Center, Clay Center, Nebraska 68933-0166

Proper distribution of immune cells in the uterus is a prerequisite for successful implantation and subsequent placentation, but biochemical signals that govern such events have not been well characterized. In the present study, the cDNA of a chemokine, interferon (IFN)--inducible protein 10 kDa (IP-10), was identified from a cDNA subtraction study between uterine endometrial tissues from Day 17 pregnant and Day 15 cyclic ewes. The effect of IFN- on IP-10 expression and the involvement of IP-10 in the recruitment of immune cells were then investigated. Northern blot analysis revealed that large amounts of IP-10 mRNA were present during conceptus attachment to maternal endometrium and early placentation. IP-10 mRNA was localized to monocytes distributed in the subepithelial stroma of pregnant but not cyclic uteri. This finding was supported by the discovery of IP-10 mRNA expression in monocytes but not in lymphocytes, uterine epithelial cells, or stromal cells. Moreover, the expression of IP-10 mRNA by the monocytes was stimulated by IFN-, IFN-, and IFN- in a dose-dependent manner, but the expression of IP-10 mRNA by the endometrial explants was most stimulated by IFN-. In a chemotaxis assay, migration of peripheral blood mononuclear cells was stimulated by the addition of IFN- stimulated-endometrial culture medium, and the effect was significantly reduced by neutralization with an anti-IP-10 antibody. These results suggest that endometrial IP-10 regulated by conceptus IFN- regulates recruitment and/or distribution of immune cells seen in the early pregnant uterus.

¹ This work was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan, by the Program for Promotion of Basic Research Activities for Innovative Bioscience (BRAIN), and by a Grant-in-Aid of Recombinant Cytokines Project from the Ministry of Agriculture, Forestry and Fisheries of Japan. K.N. was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.

² Correspondence: Kazuhiko Imakawa, Laboratory of Animal Breeding, Faculty of Agriculture, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. FAX: 81 3 58418180; akaz{at}mail.ecc.u-tokyo.ac.jp

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