Abstract

IgE-specific reverse plaque assay for the direct comparison of the IgE and IgG antibody responses was established and the method was employed for the assessment of the activity of IgE class-specific suppressor factor (IgE-TsF). In the in vitro culture system, the addition of IgE-TsF to DNP-KLH-primed spleen cells inhibited an antigen-induced increase of IgE-producing cells but did not show any effect on the IgG or IgM responses. Absorption of IgE-TsF with IgE-producing hybridoma cells removed the suppressor activity but IgM-producing hybridoma cells did not absorb the suppressor activity. The suppressor activity of IgE-TsF was removed by murine IgE-conjugated Sepharose column but not by IgM-, IgG-, or human IgE-conjugated column. The suppressor activity was eluted from IgE-column with glycine-HCI buffer, pH 3.2, or acetate buffer, pH 4.0, and the suppressor factor eluted from IgE-column was reabsorbed by anti-H-2d conjugated column. The results showed that IgE-specific suppressor factor was composed of the binding sites for IgE molecules and the H-2 gene products.

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