The main aim of the original proposal was the coordination of twelve European laboratories for the best possible use of a unique library of 150 characterized monoclonal antibodies (mABs) against the nicotinic acetylcholine receptor (AChR) in multi-disciplinary studies on the AChR and on the neurological disease myasthenia gravis (MG), which is caused by the destructive activity of anti-AChR autoantibodies.
The opportunity now arises to expand this network by the addition of three selected research groups with extensive and complementary expertise in AChR and MG research, to the:
A. Study of neuronal AChRs, using currently available and new tailor-made mAbs (M. Skok, V. Skok, Kiev; Tzartos, Athens). A small part of the original project involved studies of the neuronal AChR. We now propose to expand this work by the thorough investigation of ganglionic AChRs using currently available and novel mabs, Specifically:
Synthetic peptides will be initially used to elicit oligoclonal antibodies against the ligand-binding sites of the neuronal AChRs, against regions conserved between the muscle and neuronal AChRs and against recombinant AChR subunits. Those peptides and recombinant fragments which induce the production of antibodies binding to intact AChRs will be subsequently selected for the production of specific mabs.
The new antibodies, as well as some of those currently available which cross-react with neuronal AChR, will be used in electrophysiological and biochemical studies of AChRs in the neurons of the rat superior cervical ganglia and the guinea-pig submucous plexus to study the AChR subunit composition and the function of putative ligand-binding sites. They will also be used in the neurons of other autonomic ganglia and plexuses to study AChR diversity in the autonomic nervous system.
B. Study of AChRs on thymocytes and peripheral blood lvmphocvtes in MG Szelenyi, Budapest). This part aims to determine the expression function and degradation of AChRs on thymocytes and PLBs from healthy individuals and MG patients. The work will involve the characterization of the AChR expressed on lymphocytes at various stages of maturation. The background of the reduced AChR expression of PLBs from MG patients will also be studied. International collaboration is obviously a prerequisite for these studies.
Funding SchemeCSC - Cost-sharing contracts