Antigen-gelonin conjugates. Preparation and application in experimental myasthenia gravis.

Abstract

The antigen-specific immune suppression by gelonin-antigen conjugates was tested in two different systems: (i) the horseradish-peroxidase-stimulated T-cell proliferation in vitro and (ii) in vivo with experimental autoimmune myasthenia gravis (EAMG) in the rat. For this, the phytotoxin gelonin, a glycoprotein from Gelonium multiflorum, was purified and linked to the respective antigens. For the in-vitro assay a lymph node cell suspension from rats immunized with horseradish peroxidase was cultured in the presence of this protein and proliferation was measured by [3H]thymidine uptake. In-vitro proliferation was significantly inhibited by adding gelonin-horseradish peroxidase conjugates. The therapeutic effects of antigen-gelonin conjugates were tested in the rat model EAMG. For these experiments rats were immunized with purified nicotinic acetylcholine receptor from electric fish in order to develop EAMG. The success of the immunization was monitored by the change in physical performance tests, the change in anti-acetylcholine receptor antibody titer, and by the change in the number of ionic endplate channels using a novel electrophysiological method. The latter method permits a very accurate assay of functional damage of acetylcholine receptor at the endplate and correlates well with the clinical severity of the disease. Rats were conventionally immunized with acetylcholine receptor from electric fish. After the onset of EAMG as measured by physical performance tests and rise in antibody titer a group of the animals was injected with an acetylcholine receptor-gelonin conjugate and this treatment was repeated seven days later. The loss in functional acetylcholine receptor was significantly smaller in the therapy group than in the untreated EAMG group.(ABSTRACT TRUNCATED AT 250 WORDS)