Epitope mapping of 10 monoclonal antibodies against the pig analogue of human membrane cofactor protein (MCP)

Lastra, José Manuel Pérez de la; Berg, Carmen W. van den; Bullido, R; Almazán, Fernando; Domı́nguez, Javier; Llanes, D.; Morgan, B. Paul

doi:10.1046/j.1365-2567.1999.00732.x

Cited by 14 publications

(11 citation statements)

References 27 publications

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“…Immunohistology using a well‐characterized anti‐pig MCP monoclonal antibody 12 , 22 confirmed that MCP was abundantly distributed in pig tissues, particularly on cells of epithelial and endothelial origin (Fig. 4; Table 1).…”

Section: Discussionmentioning

confidence: 74%

Distribution of membrane cofactor protein (MCP/CD46) on pig tissues. Relevance to xenotransplantation

1999

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Membrane cofactor protein (MCP; CD46) is a 50-60 000 MW glycoprotein, expressed on a wide variety of cells and tissues in man, which plays an important role in regulating complement activation. Human MCP has also been shown to be the receptor for measles virus. We have recently identified the pig analogue of MCP and demonstrated that pig MCP has cofactor activity for factor I-mediated cleavage of C3b when these components are derived either from pig or human. As a consequence, pig MCP is an efficient regulator of the classic and alternative pathways of human and pig complement. In order to define the potential importance of MCP in protecting against complement activation in the pig, we have conducted a comprehensive survey of its distribution in pig cells and organs. As in humans, MCP in the pig is broadly and abundantly distributed. Pig MCP is highly expressed on all circulating cells, including erythrocytes, in contrast to its absence on human erythrocytes. Multiple isoforms of MCP are found on cells and in tissues, probably representing products of alternative splicing analogous to those found in man. MCP is abundantly expressed throughout all tissues examined with particularly strong staining on the vascular endothelium. Connective tissue elements within liver and testis are also strongly stained by anti-pig MCP antibodies. Pig MCP is expressed only weakly on skeletal muscle cells and expression is absent from smooth muscle cells in the lung and vessel walls, sites at which human MCP is expressed. Of particular note, MCP is not expressed in B-cell areas of the germinal centres of lymph nodes.

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Section: Discussionmentioning

confidence: 74%

Distribution of membrane cofactor protein (MCP/CD46) on pig tissues. Relevance to xenotransplantation

1999

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“…Clustering of the different mAbs based on a mAb-competition assay was previously shown for other proteins using a combination of biotinylated and non-biotinylated mAbs [44,45]. An excess of unbiotinylated mAbs was added to the cells and were allowed to bind to surface-expressed Sn for 60 min at 37 °C.…”

Section: Resultsmentioning

confidence: 99%

Development and Characterization of New Species Cross-Reactive Anti-Sialoadhesin Monoclonal Antibodies

et al. 2016

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Sialoadhesin (Sn) is a surface receptor expressed on a subset of macrophages in steady state conditions. During inflammation and diseases, Sn is highly upregulated on macrophages and blood monocytes. Therefore, therapies using monoclonal antibodies (mAbs) to target Sn-positive (Sn + ) cells are a potential strategy for targeted treatment. It has been shown that Sn internalizes after binding with a mAb, though it is not clear whether this is species-specific. In this study, new Sn-specific mAbs were developed and analyzed for cross-reactivity between species. In addition, the newly developed mAbs were compared to mAbs used in previous research for their epitope recognition and other Sn-specific characteristics. Both species-specific and cross-reactive antibodies could be identified. Furthermore, sialic acid-binding of red blood cells (RBC) could be inhibited with mAbs recognizing different epitopes and all mAb showed internalization of Sn. The newly developed mAbs can be used as novel tools for Sn research and further analysis of Sn internalization in different species.

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“… 20 Since expected results were obtained in the CHO system, similar expression trials were next carried out with two swine endothelial cell lines, SEC‐E and PAE. These cell lines expressed pig MCP, 19 which was detectable only with specific mAbs 19,30 Many clones expressing the four human MCP variants were successfully established ( Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

“…What works as an alternative receptor in SEC‐E cells is an intriguing issue. Although pigs have their own MCP on most tissue/organs, 19,30 , 32 the pig MCP does not act as a MV receptor (data not shown). It has been reported that ‘scatter‐like’ transdifferentiation was observed in endothelial and epithelial cells including hepatic cells treated with hepatocellular growth factor (HGF) 33,34 .…”

Section: Discussionmentioning

confidence: 96%

Molecular remodelling of human CD46 for xenotransplantation: designing a potent complement regulator without measles virus receptor activity

et al. 2000

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SUMMARYIn pig-to-human discordant xenotransplantation, human complement (C) is a major barrier to long survival of xenografts. The current idea on how to cope with this barrier is that human complement regulatory proteins are forcibly expressed on xenografts to serve as safeguards against host C-induced hyperacute rejection of xenografts. Co-expression of decay-accelerating factor (DAF) (CD55) and membrane cofactor protein (MCP) (CD46) would be the ®rst choice for this trial, because most of the human cells are protected from C-mediated damage by two different modes with these two kinds of Cregulators. Many problems have arisen, however, for MCP expression on grafts. (i) MCP acts as a measles virus receptor, which may function to render donor pigs measles virus (MV) sensitive. (ii) MCP signals immune suppression which causes devastation of the recipient's immune responses. (iii) MCP exerts relatively low self-protective activity against C compared with other cofactors; development of more ef®cient forms is desirable. (iv) Grafts with a high expression level of MCP are dif®cult to produce. In this study, we made a number of cDNA constructs of MCP, expressed them on swine endothelial cell lines, and tested cell-protective potency and MV susceptibility. The short consensus repeat 1 (SCR1)-deleted MCP with glycosyl phosphatidylinositol (GPI)-anchored form (D1MCP-PI) of MCP was found to be most suitable for the purpose of overcoming these problems. However, it was also found that MV induces two modes of cytopathic effect (CPE) on swine endothelial cells, either MCPdependent or -independent. Here, we discuss these two points which will be raised through study of MCP-transgenic animals.

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Epitope mapping of 10 monoclonal antibodies against the pig analogue of human membrane cofactor protein (MCP)

Cited by 14 publications

References 27 publications

Distribution of membrane cofactor protein (MCP/CD46) on pig tissues. Relevance to xenotransplantation

Distribution of membrane cofactor protein (MCP/CD46) on pig tissues. Relevance to xenotransplantation

Development and Characterization of New Species Cross-Reactive Anti-Sialoadhesin Monoclonal Antibodies

Molecular remodelling of human CD46 for xenotransplantation: designing a potent complement regulator without measles virus receptor activity

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