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The switch from IgM to IgG secretion in single mitogen-stimulated B- cell clones

J. Andersson,A. Coutinho,F. Melchers

Published 1978 in Journal of Experimental Medicine

ABSTRACT

The frequency of mitogen-reactive B cells yielding an IgG plaque- forming cell (PFC) response has been determined in vitro by limiting dilution analysis under culture conditions which allow every growth- induced B cell to grow and mature into a clone of Ig-secreting cells. The frequencies of lipopolysaccharide (LPS)-and lipoprotein-reactive precursors for IgG-secreting cells in the spleen of 6--8 wk old C3H/Tif and of C57BL/67 mice were found to be between 1 in 30 and 1 in 40 B cells and, therefore, only one tenth of the frequencies of mitogen- reactive precursors of clones secreting IgM. All IgG-secreting cells developed by switching in clones which previously contained IgM- secreting cells. This was shown in two experiments where the total number of mitogen-reactive precursor yielding IgM-secreting cell clones was limited such that 82 or 90% of all responding cultures originated from one precursor. Thus, of 480 cultures in the first and 720 cultures in the second experiment, 86 and 98 cultures were found positive, yielding IgM-secreting cells at day 5. When the same cultures were assayed at day 7 for IgG-secreting cells 9 and 10 cultures were found positive. All 19 cultures with IgG-secreting cells previously had contained IgM-secreting cells. The probability that IgG-secreting cells and IgM-secreting cells would have arisen from independent precursors can be calculated using Fisher's exact test of independence. For the two experiments those probabilities are 3.4 X 10(-7) and 4.0 X 10(-9). Since we have previously shown that each cell in a mitogen-stimulated, growing B-cell clone divides, and that each dividing cell secretes Ig, we conclude from these experiments that the large majority--in our experiments all--of the IgG-secreting cells in mitogen-stimulated B- cell clones develop by switch from IgM-secreting cells. IgG-secreting cells develop either early or late during growth of a single IgM- secreting cell clone. The switch to IgG secretion, therefore, is not fixed in the time of clonal growth after mitogenic stimulation.

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