Sheep erythrocytes, artificially coated with purified antibody specific for mouse serum albumin (anti-MSA Ab-E) were agglutinated and lysed by mouse serum albumin. Mouse liver cells, plated with anti-MSA Ab-E formed hemolytic plaque in a modified plaque-forming cell (PFC) assay. The same cells tested with anti-MSA Ab-E did not rosette in modified immunocyto-adhesion assay. Thus, liver cells secreted molecules (albumin) which were not detectable on the cell membrane. This PFC assay with Ab-E is probably a general method for enumerating antigen-secreting cell.

Agglutination↗

PubMed1975

Enumerating immunoglobulin-secreting cells among peripheral human lymphocytes. A hemolytic plaque assay for a B cell function.

Cells bearing immunoglobulin (Ig) can be enumerated by current methods, but cells secreting Ig (with unknown antibody specificity) can not. By using a reverse plaque-forming cell assay, we counted the Ig-secreting cells in the blood of nonimmunized individuals. Isolated human lymphocytes were plated in gels together with erythrocytes coated with purified antibody to human Ig. After treatment with anti-Ig antibody and complement, the Ig-secreting cells formed hemolytic plaques. In 10 normal individuals, 1.2 +/- 0.4% of the plated mononuclear cells formed plaques; in an individual tested on 7 different days during 3 months, 1.1 +/- 0.2% of the cells formed plaques. From the number of mononuclear cells in the blood, we calculated 22 +/- 8 Ig-secreting cells per mm3 of blood. Enumerating Ig-secreting cells is a new direct assay for a B cell function and need not correlate with quantitating serum Ig. Thus, this assay should be useful for studying Ig-secretion in normal and altered states.

Animals↗

PubMed1973

Detection of the e14 and e15 rabbit allotypic specificities by immunodiffusion in PEG agar.

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Agar↗