Haemolytic Disease of the Foetus and Newborn (HDFN) and Foetal and Neonatal Alloimmune Thrombocytopenia (FNAIT) are the most clinically relevant alloimmune disorders of pregnancy caused by maternal alloimmunisation to paternally derived foetal red blood cell (RhD) and platelet antigens (HPA-1a) respectively. Recombinant Fc-modified antibodies have been designed as inert potential biotherapeutics to compete with maternal alloantibodies and reduce foetal mortality. Fc-modified anti-D (Fog1G1 Δnab) and anti-HPA-1a (B2G1Δnab & B2G1Δnac) have been evaluated for their materno-foetal transport capacity using human placental models. For future in vivo efficacy, Fc-modified antibodies should transport at similar rates to wild-type antibodies (Fog1G1 and B2G1). The placental perfusion model showed that the Δnab mutation appeared to lower the transport capability of anti-D and anti-HPA-1a across the placenta. In a Human Umbilical Endothelial Vein Cell (HUVEC-c) cell culture model, transport of HPA-1a was favoured in a basal to apical direction and was statistically significant at hours 12 and 24 (p=0.002 & p=0.010 respectively). The relative order of transport was B2G1Δnac > B2G1 > B2G1Δnab implying the Δnac mutation enhances transport across the foetal endothelium. Since approximately 40% of RhD negative women give birth to RhD negative babies, these women currently receive anti-D prophylaxis unnecessarily. Foetal DNA was successfully extracted from maternal plasma and genotyped for foetal RhD status using Real-Time PCR. Foetal genotyping results revealed 96% and 98% concordance with cord blood serology for maternal blood samples taken at booking (~16 weeks) and at 28 weeks gestation respectively. Two-dimensional Difference in Gel Electrophoresis (2-D DiGE) was used to evaluate the normal placental proteome of syncytiotrophoblast membrane particles (STBMs) generated from placental perfusion. Eleven differentially expressed protein species were identified when comparing different STBM samples. Future work aims to compare the normal placental proteome with the proteome of placentas from complicated pregnancies (e.g. PE, IUGR, PTL and Trisomies 13, 18 and 21) to discover potential biomarkers for screening.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:553835 |
| Date | January 2011 |
| Creators | Porter, Charlene |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=167960 |
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