Complement regulatory proteins are expressed at low levels in embryonic human, wild type and transgenic porcine neural tissue

• Published in: Xenotransplantation (Københaven) 2004-01, Vol.11 (1), p.60-71
• Main Authors: Harrower, T. P., Richards, A., Cruz, G., Copeman, L., Dunnett, S. B., Barker, R. A.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified - embryology; Animals, Genetically Modified - genetics; Antigens, CD - genetics; Antigens, CD - metabolism; Brain - embryology; CD55 Antigens - genetics; CD55 Antigens - metabolism; CD59; CD59 Antigens - metabolism; Cells, Cultured; complement; Complement Inactivator Proteins - metabolism; decay accelerating factor; Embryo, Mammalian - metabolism; Humans; Immunohistochemistry; Membrane Cofactor Protein; Membrane Glycoproteins - genetics; Membrane Glycoproteins - metabolism; neural transplantation; Swine - embryology; transgenic pigs; xenotransplantation
• ISSN: 0908-665X; 1399-3089
• DOI: 10.1111/j.1399-3089.2004.00084.x

:  Allotransplantation of human foetal neural tissue for neurodegenerative disorders has been shown to provide clinical benefit but is limited by a number of issues including donor supply. The use of porcine foetal tissue as an alternative source of cells is being investigated but xenotransplants survive poorly as a result of immunological rejection, which may involve complement. In this study we investigated the expression of the membrane‐bound complement regulatory proteins – decay accelerating factor (DAF), membrane co‐factor protein (MCP) and CD59 in embryonic neural tissue. Cells were derived from human foetuses, wild‐type porcine foetuses and porcine foetuses transgenic for human complement regulatory proteins and analysed using flow cytometry and immunocytochemistry. Functional assessment of human complement regulatory protein expression in transgenic porcine tissue was assessed by C3b deposition and cell survival on exposure to human complement. Human and wild‐type porcine foetal neural tissue expressed moderate levels of MCP and CD59 but low or no levels of DAF. Neural tissue from porcine foetuses transgenic for human MCP (E174) expressed the transgene but failed to significantly inhibit human C3b deposition compared with non‐transgenic tissue. In contrast, foetal neural tissue from two different human DAF transgenic pig lines (A74 and E71) known to express high levels of human DAF on endothelial cells, failed to express significant levels of human DAF in foetal neural tissue. Complement regulatory proteins such as MCP and CD59 are expressed in the human and wild‐type embryonic brain but in contrast, DAF is expressed at very low levels. Pigs transgenic for human DAF express very low levels of human DAF on embryonic neural tissue. In pigs transgenic for human MCP, the transgene is expressed at similar levels to that in human embryonic neural tissue but at an insufficient level to prevent activation of the complement cascade. Thus alternative approaches to reducing complement activation by xenografted neural foetal tissue will be required if this process proves to be important in the rejection process.