In primates, prenatal transfer of IgG from mother to offspring occurs predominantly across the placenta. Although a number of Fcgamma-receptors and IgG binding proteins have been detected in human placental tissue, an involvement of any of these receptors in IgG transport across the syncytiotrophoblast remains to be demonstrated. Therefore, we investigated the mechanism of IgG transcytosis in trophoblast-derived BeWo cells. BeWo cells were not only found to express the MHC class I-related IgG Fc receptor, human FcRn, but also specifically bound fluorescein isothiocyanate (FITC)-labeled human IgG (FITC-hIgG) at the apical surface at mildly acidic pH. The cells preferentially transcytosed FITC-hIgG from the apical to the basolateral side when compared to the fluid-phase marker FITC-dextran and to FITC-hIgG transcytosis in the opposite direction. However, endocytosis of FITC-hIgG at the apical plasma membrane at physiological pH required the continuous presence of FITC-hIgG at concentrations similar to those present in the maternal circulation. These results suggest a mechanism by which IgG is internalized by BeWo cells via fluid-phase endocytosis. Tight binding of IgG to hFcRn may then occur in acidic endosomes, followed by selective sorting into the transcytotic pathway. Thus, the main function of this receptor is to prevent entry of IgG into the degradative pathway in lysosomes.

In primates, prenatal transfer of IgG from mother to offspring occurs predominantly across the placenta. Although a number of Fcgamma-receptors and IgG binding proteins have been detected in human placental tissue, an involvement of any of these receptors in IgG transport across the syncytiotrophoblast remains to be demonstrated. Therefore, we investigated the mechanism of IgG transcytosis in trophoblast-derived BeWo cells. BeWo cells were not only found to express the MHC class I-related IgG Fc receptor, human FcRn, but also specifically bound fluorescein isothiocyanate (FITC)-labeled human IgG (FITC-hIgG) at the apical surface at mildly acidic pH. The cells preferentially transcytosed FITC-hIgG from the apical to the basolateral side when compared to the fluid-phase marker FITC-dextran and to FITC-hIgG transcytosis in the opposite direction. However, endocytosis of FITC-hIgG at the apical plasma membrane at physiological pH required the continuous presence of FITC-hIgG at concentrations similar to those present in the maternal circulation. These results suggest a mechanism by which IgG is internalized by BeWo cells via fluid-phase endocytosis. Tight binding of IgG to hFcRn may then occur in acidic endosomes, followed by selective sorting into the transcytotic pathway. Thus, the main function of this receptor is to prevent entry of IgG into the degradative pathway in lysosomes.

The human homologue of FcRn, an IgG Fc receptor expressed in rat villous syncytiotrophoblasts, might be involved in IgG transfer from the maternal to the fetal circulation. However, because the receptor does not bind IgG at the physiological pH of the maternal blood (pH 7.4), FcRn is probably not involved in the initial uptake of IgG. A role in IgG internalization has been suggested for placental alkaline phosphatase (PLAP), which is highly expressed on the apical surface of syncytiotrophoblasts. To determine whether PLAP does indeed have a role in IgG uptake, we analysed the ability of PLAP to bind, internalize and transcytose IgG in BeWo choriocarcinoma cells endogenously expressing the protein, or in Madin-Darby canine kidney (MDCK) cells transfected with the PLAP cDNA. Although PLAP expression in MDCK cells resulted in increased IgG binding to intact cells, binding was not correlated with the level of PLAP expressed in the different cell lines. Furthermore our findings do not support a role for PLAP in IgG endocytosis or transcytosis.

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Small signal Surface Photo-voltage (SPV) measurement techniques have been applied to monitor ion implants typical of those used for layer-transfer SOI processes. This SPV wafer mapping technique was investigated for sensitivity to dose, implant uniformity, and repeatability for hydrogen and helium implants into (100) silicon wafers through a 1450Aå surface oxide. A normalized sensitivity of approximately 1.2 for the investigated cases was observed. A demonstrated repeatability of less than 0.5% 1-sigma standard deviation was measured for a multi-day period on hydrogen implanted wafers. The small signal SPV metrology technique presented allows in-line SPC control of the critical ion implant step within the SOI wafer manufacturing process.