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Enumeration of Trypanosoma cruzi-specific antibody-secreting cells by solid-phase immunoenzymatic technique
1Departamento de Imunologia, Universidade de São Paulo, Brasil.
A specific and sensitive enzyme-linked immunoassay is applied for enumeration of cells secreting specific antibodies to Trypanosoma cruzi antigens. Spleen cells from immunized mice are incubated in antigen-coated plastic tissue culture plates. Individual antibody-producing cells secrete antibody which binds to the antigen at close vicinity of the cells. The areas of bound antibodies are demonstrated by an enzyme-linked antibody assay as dark round spots, which can be easily enumerated. This assay allows the enumeration of specific antibody-secreting cells to parasite antigens, overcoming the failures to develop conventional plaque-forming cells assays to complex antigens.
A specific and sensitive enzyme-linked immunoassay is applied for enumeration of cells secreting specific antibodies to Trypanosoma cruzi antigens. Spleen cells from immunized mice are incubated in antigen-coated plastic tissue culture plates. Individual antibody-producing cells secrete antibody which binds to the antigen at close vicinity of the cells. The areas of bound antibodies are demonstrated by an enzyme-linked antibody assay as dark round spots, which can be easily enumerated. This assay allows the enumeration of specific antibody-secreting cells to parasite antigens, overcoming the failures to develop conventional plaque-forming cells assays to complex antigens.