IN VITRO
JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
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Protein transport across the in vitro perfused human placenta.

PROBLEM: Placental transport of various proteins present in human serum, such as immunoglobulins (IgG, IgA), specific anti-tetanus IgG (anti-TT-IgG), and tetanus toxoid-antigen (TT-AG), was investigated. In addition, the transport of IgG modified with biotin (IgG-BT) and 14C-bovine serum albumin (14C-BSA, a permeability marker for macromolecules), was assessed.

METHOD OF STUDY: During the perfusion of an isolated cotyledon from human term placenta the perfusate was recirculated on both maternal and fetal sides. After an initial stabilisation phase of 2 hr (control phase), media on both sides were exchanged and perfusion was continued comparing two different conditions (experimental phase). In the first group (control experiments [A, n = 3]), no test proteins were added during the experimental phase (4-6 hr). In the second group (B, n = 5), during the experimental phase (6 hr) the maternal perfusion medium contained IgG (Sandoglobuline, 6-10 g/L), anti-TT-IgG (21-25 mg/L), TT-AG (0.19-0.24 mg/L), and IgA (0.13-0.19 g/L). IgG-BT (2 g/L) and 14C-BSA (30-40 nCi/ml) were added to the medium on the maternal side. IgGs and TT-AG were determined by specific enzyme-linked immunosorbent assay.

RESULTS: Both groups showed stable metabolic conditions with constant rates of glucose consumption, lactate production, and hormone (human chorionic gonadotropin, human placental lactogen) release observed throughout the experiment. Washout levels of endogenous IgG and IgA observed in the maternal circuit at the end of the control period were 5 and 1000 times higher than in the fetal circuit. In the experimental phase these levels remained constant at 50-80% of control levels with no change in the last 4 hr of perfusion (group A). In group B, with addition of extra proteins, trace amounts of IgG-BT, IgA, and 14C-BSA were detectable in the fetal circuit within 1 hr, with no significant further increase in circulating levels in the following 4 hr of the perfusion. In contrast, the detection of IgGs in the fetal circuit was delayed by 2 hr; thereafter, a continuous linear increase was observed for all IgGs. TT-AG in fetal perfusate was below the detection limit. TT-AG was found on the fetal side only after ultrafiltration of samples obtained at the end of the experiment. For permeability comparison, the ratio between concentrations on the fetal and maternal side multiplied by 100 ([F:M] x 100), as detected after 6 hr of perfusion, was assessed (n = 5, mean +/- SD). Labelling of IgG with biotin (IgG-BT) reduced its placental transfer by a factor 10 (0.04 +/- 0.01) when compared with the natural IgG (0.49 +/- 0.08) or the specific antibody (anti-TT-IgG). The relative fetal-to-maternal ratio found for TT-AG (0.48 +/- 0.12) was similar to anti-TT-IgG (0.46 +/- 0.11), and approximately 4 and 50 times that of 14C-BSA (0.12 +/- 0.03) and IgA (0.01 +/- 0.01), respectively. Considering that the molecular weights of TT-AG and anti-TT-IgG were at least twice that of BSA and similar to IgA, the difference in transfer suggests a specific mechanism of transport.

CONCLUSIONS: Compared with other proteins there is a significantly increased transfer of IgGs across the in vitro perfused human placenta from the maternal to the fetal side, indicating a specific transport mechanism. The similarity in transfer of anti-TT-IgG and tetanus antigen may suggest the transport as antibody-antigen complex.

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