A combined staining, direct immunofluorescence horseradish peroxidase technique is presented for demonstrating both the class of immunoglobulin and the antigen specificity of antibody within single cells. First, a cryostat secretion is incubated with fluorescein labeled antibody against the heavy chain of IgG, IgM or IgA, and is photographed. Then the same section is incubated with peroxidase labeled antigen and reached with diaminobenzidine. The previously fluorescent cells are relocated and the presence or absence of peroxidase reactivity in the same cells is determined. We have used this technique to study (a) the cells in experimental allergic neuritis that spcifically bind the P2 basic protein from peripheral myelin and (b) the cells in experimental allergic encephalomyelitis that specifically bind the encephalitogenic basic protein from central myelin. In rabbits with either disease, the mononuclear cells in lymph nodes and nervous system target organs which bound peroxidase labeled basic protein antigens contained immunoglobulin of the IgG, but not of the IgM or IgA class.
|Original language||English (US)|
|Number of pages||6|
|Journal||Journal of Histochemistry and Cytochemistry|
|State||Published - Jan 1 1975|
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