标题：Antigen-Secreting Cells: Enumeration of Immunoglobulin-Allotype-Secreting Cells in Nonimmunized Rabbits by Means of Hybrid-Antibody-Coated Erythrocytes in a Reverse Hemolytic Plaque Assay
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1974
卷号：71
期号：4
页码：1229-1233
DOI：10.1073/pnas.71.4.1229
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：A reverse hemolytic plaque-forming cell (PFC) assay for the enumeration of immunoglobulin-(Ig)secreting cells in nonimmunized rabbits was developed by using erythrocytes coated with anti-Ig antibody. These indicator cells lysed after reacting with the secreted Ig antigen and complement. The anti-Ig antibodies used were specific for the b4 or b5 allotypic specificities, which are antigenic determinants on the kappa chain of rabbit Ig and controlled by alleles at the b locus. We coated erythrocytes with anti-b4 or anti-b5 antibody, using hybrid antibody with dual specificity to sheep red blood cells and to the b4 or b5 kappa chain. The coated erythrocytes were plated with cells from various lymphoid organs of nonimmunized rabbits. These lymphoid cells formed allotype-specific plaques, i.e., the cells of b4 homozygous rabbits formed hemolytic plaques with anti-b4 but not with anti-b5-coated erythrocytes and vice versa. In eight nonimmunized rabbits, 0.06-0.3% of the spleen cells (590-2900 PFC per 106 cells) secreted the b locus Ig allotypes. In three nonimmunized rabbits, 320-590 PFC per 106 cells were counted in the lymph node or bone marrow; 26-40 PFC per 106 cells were detected in the thymus. Since the b4 and b5 specificities are present on approximately 90% of the serum Ig, our results indicate that less than 1% of the lymphoid cells in nonimmunized rabbits secrete Ig at any one time. By this reverse PFC assay, Ig-secreting cells were for the first time detected as antigen-secreting cells. This assay can be applied to the detection of cells secreting other antigens.
