Polyclonal Antibody Secretion Induced in Human Mixed Lymphocyte Cultures

Antibody secreting B cells were measured as plaque forming cells (PFC) in a hemolysis-in-gel assay using fluorescein isothiocyanate (FITC) coupled SRBC as targets or protein A coupled SRBC as targets and developing antisera. Peak antibody secretion occurred on day 5 and the highest number of PFC was seen when mitomycin treated stimulator cells: responder cells were used in a ratio of 1:4. The number of PFC in MLC was not correlated to the DNA synthetic response. Antibody secretion in MLC was found to be of IgM, IgG and IgA classes. Significant numbers of PFC in MLC from blood lymphocytes were detected with the protein A technique, but not using FITC-SRBC targets. Compared to spleen cells, fewer PFC were stimulated in blood lymphocytes. B cells alone, enriched by rosetting of T cells, did not respond by antibody secretion or DNA synthesis in MLC. When Lipopolysaccharide (LPS) was added to MLC an additional effect was seen on the number of PFC which may indicate that distinct B cell subpopulations are activated in MLC by LPS.