Use of a Fluorescein-labeled Sonically Disrupted Bacterial Antigen to Demonstrate Antibody-producing Cells.

Eveland, Warren C. (University of Michigan, Ann Arbor). Use of a fluorescein-labeled sonically disrupted bacterial antigen to demonstrate antibody-producing cells. J. Bacteriol. 88:1476-1481. 1964.-Cells obtained by primary tissue culture of the spleens of chickens immunized with sonically disrupted Escherichia coli O111 organisms were stained with a fluorescein-labeled homologous antigen by use of direct immunofluorescent methods. Brilliant staining of the cytoplasm in cells from immunized birds appeared to be diffuse in certain cells and rather globular in others. In contrast, cells from nonimmunized birds showed no staining at all. The cells involved in the specific reaction appeared to be those of the lymphocyte-monocyte-plasma cell types, as shown when stained by the May-Grunwald-Giemsa method. Preparations stained by the methyl green-pyronin technique revealed an increase in the pyrinophilic cells in the preparations from the immunized birds, thus demonstrating increased amounts of ribonucleic acid in these cells, which in turn is consistent with the presence of antibody globulin. Specificity of the reaction was confirmed also by (i) staining antibody-coated E. coli O111 organisms with the conjugate, (ii) precipitin reaction with specific antibody, and (iii) specific agglutination with circulating antibody from the immunized birds.