Limiting dilution analysis of dsDNA binding production by PerC B-1a B cells (original) (raw)

PLOS ONE, 2015

Abstract

<p>Frequency of peritoneal cavity B-1a cells expressing dsDNA-binding BCR by genotype. PerC B-1a cells were sorted and cultured under polyclonal stimulation for IgM secretion. The frequency of B-1a cells secreting IgM anti-dsDNA <i>in vitro</i> was estimated by LDA. <b>A)</b> Histogram bars show the frequency, calculated by the Poisson distribution, of culture supernatants containing IgM binding to dsDNA. <b>B)</b> O.D. values of individual culture with variable numbers of B-1a cells. <b>C)</b> Evaluation of anti-dsDNA binding of culture supernatants containing 10,000 B-1a cells and similar amount of total IgM per culture. <b>D)</b> Binding to non-related antigen (phosphorylcholine—PC) using the same culture supernatants to control for the specificity of the assay. Significantly different from WT: *, P ≤ 0.05; **, P ≤ 0.01.</p

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