A.8 Placenta (1997), Vol. 18

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EFFECT OF SERA FROM PREECLAMPTIC PATIENTS UPON THE APPEARANCE OF ADHESION MOLECULES ON CULTURED HUMAN TROPHOBLASTIC AND ENDOTHELIAL CELLS. Emiko Abe, Keiichi Matsubara, Hiroshi Ochi, Hiroyuki Kitagawa, Masaham Ito, Keizo Okall), Kenji Kamedacll, Department of Obstetrics and Gynecology, and Central Research Laboratory(l), School of Medicine, Ehime University, Ehime, Japan

[Objectives] Activation of vascular endothelial cells (E&l is thought to be one of the important pathophysiologic conditions for the occurrence of preeclampsia. It is possible that trophoblastic cells (TCs) play a role similar to ECs in the placenta and are activated like ECs in preeclampsia. We investigated the appearance of adhesion molecules on cultured cells stimulated by sera from preeclamptic patients. [Materials and Method] Villi which were obtained by artificial abortions (6-8 gestational weeks) were treatrd with trypsin and DNase. The cells were separated by Percoll and cultured in gelatin coated dishes for 5-6 days. ECs were collected from the umbilical vein of normally delivered placenta using Jaffe’s method, and were cultured in collagen coated dishes for 5-7 days. Medium containing sera preeclamptic patients and normal pregnant women were addcd 10 the cultured ceils and incubated for 0, 5, SO, 60, 120 mitt, 12. and 24 hours, Anti E-selectin, ICAM-1, and VCAM-1 antibodies conjugated to FITC were incubated with the cells following trypsin treatment The expression levels of the adhesion molcules were measured by flow cytometer. [Results] No significant change was observed in E- selection and VCAM-1 expression in each groups. The expression level of ICAM- 1 increased with time. Preeclamptic sera increased significantly ICAM- compared with normal pregnant women’s sera. [Conclusion] ICAM- appeared on both TCs and ECs. ICAM- may play a role in the development.

IMMUNOHISTORICAL LOCALIZATION OF INSULIN-LIKE GROWTH FACTOR-I (IGF-I), IGF-I REVELER AND IGF-BINDING PROTEIN- 1 ~GFBP- 1) IN THE HUMAN ANCHORING VILLI. SFunayama, T. Iwashita*, H.Funayama**, Y.Takeda*, T. Takayama**, P. Kaufmann*** OB/GYN Shiseikai-daini Hospital, Tokyo Women’s Medical College*, Tokyo Medical College, Japan**, Anatomy RWTH Aachen, Germany***

[Aim] The anchoring villus is a part of placental tissue that contains trophoblast ceils invading into the maternal decidua. We investigated the Immunohistochemical localization of IGF-I, IGF-I receptor(IGF-I R.) and IGFBP-lin anchoring villi obtained from human villous tissue to elucidate the physiological significance of these substances in the invasion process of trophoblast. [Method] Fresh frozen blocks were made from placentae at 6-17 weeks of gestation after legal abortion. The immunostaining was performed with the streptavidin-biotin method using monoconal anti-bodies against IGF-I, IGF-I R. and IGFBP- I, Anti-vimentin, anti-cytokeratin and MIB-lwere also used to identify cell types. [Results] IGF-I was stained faintly in the cell column while IGF-I R. was localized in the peripheral part of villi which was also stained by MIB-1 antibody in serial sections. IGFBP-1 was not seen in trophablast tissues, but stained in decidua cells. [Conclusion] Since IGF-I R. was localized in a highly proliferating part of the anchoring villi, it is suggested that IGF-I may be involved in the invading process of the anchoring villi.

Cord Blood Cytokines and Soluble Adhesion Molecules in Normal Vaginaly Delivered Neonates. Showa Aoki, Toshiyuki Hata, Kohji Miyazaki, Department of Obstetrics and Gynecology, Shimane Medical University, Izumo 693, Japan

Objective: We assessed cord blood cytokines and soluble adhesion molecules in normal vaginaly delivered neonates. Methods: Umbilical cord IL-lb, IL-lra, IL-4, IL-6, IL-8, IL-lo, TNF ct , sTNFR-I, sTNFR-II, ICAM-1, ELAM-1 were measured in 30 normal newborns appropriately grown delivered vaginally. Results: IL-l@ and IL-IO were undetectable in normal pregnancies. IL-4 was detected in 47.6% of cases. Each value measured by ELISA for IL-lra, IL-4, IL-6, IL-8, TNF a,, sTNFR-I, sTNFR-II, ICAM-1, and ELAM-1 was 2,117&2,387pg/m1, 2.4&2.0pg/ml, 38.157.lpg/ml, 9.4 *0.6pg/ml, 15.6f11.2pg/ml, 3,100~400pg/ml, 7,000* l,OOOpg/ml, 195”ll lng/ml, and 222* 134ng/ml. Conclusion: IL-1 ,& and IL-10 are not physiologic constitutes of term cord blood. A balance between proinflammatory, immunosuppressive, or growth promoting cytokine activities may play a major role in the establishment and maintenance of normal human pregnancy.

CD9 IS INVOLVED IN EXTRAVILLOUS TROPHOBLAST INVASION. T. Hiranoll, T. Higuchil), H. Fujiwaral), H. Katsuragawa2), T. Inouel), K. R. Parkl), T. Moril), and M. Maeda.?), 1lDepartment of Gynecology and Obstetrics, Faculty of Medicine and 3IChest Disease Research Institute, Kyoto University, Kyoto, 2lOtsu Municipal Hospital, OISU, Shiga, Japan.

Objective: Recently, the CD9 molecule, a 2427kD cell surface glycoprotein, has been reported to be related to cell migration and adhesion, The purpose of this study was to elucidate the physiological role of CD9 in trophoblast invasion at the fctomaternal in&ace. Methods: Immunohistochemical analysis of CD9 expression in human placentae was carried out. Then using an in vitro invasion assay. we examined the effect of anti-CD9 monoclonal antibody (mAbl on invasion of BeWo cells which express CD9. The effects of anti-CD9 mAb on hCG secretion and cell proliferation were also examined. Furthermore, we examined the effect of CD9 on invasion following treatment with mAbs against integrins a3, a5, and bl. Results: CD9 was expressed on extravillous trophoblasts, but was not detected in villous cytotrophoblasts nor in syncytiotrophoblasts. Anti-CD9 mAb enhanced the number of invading BeWo cells in a dose dependent manner without affecting cell proliferation or human chorionic gonadotropin (hCG) secretion. Anti-CD9 mAb also showed a stimulatory effect on BeWo cell invasion following treatment with mAb against integrin a.3. In contrast, it had no effect following treatment with mAt)s against integrrn a5 and bl, which were also highly expressed on extravillous trophoblasts. ConclusionzThese findings indicate that CD9 is strongly related to the invasive properties of trophoblasts during implantation and placentation, and suggest that CD9 regulates trophoblast invasion via integrin a5bl.