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The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
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Effect of growth and differentiation stimuli on the development of antigen-responsive B cells in fetal liver

J M Teale1, K S Landreth

  • 1Department of Microbiology, University of Texas Health Science Center, San Antonio 78284.

Cellular Immunology
|December 1, 1988
PubMed

The development of the B cell immune repertoire was studied using an in vitro fetal organ culture system. In order to analyze the mechanism by which B cell precursors clonally expand and diversify, fetal lymphoid tissues were incubated in the presence of several factors known to influence B cell differentiation: IL-1, IL-2, WEHI-3 culture supernatant containing IL-3, and a factor from a cyclic neutropenia patient (CNF). By analyzing the effect of exogenous factors on the frequency of antigen-responsive B cells, the ability of the factor to either inhibit or enhance clonal expansion was determined. It was found that the addition of IL-1, WEHI-3 supernatant, or CNF increased the frequency of DNP-responsive B cells suggesting an enhancement of clonal expansion. IL-2, on the other hand, did not alter the frequency of antigen-responsive B cells. The effect of added factors on the kinetics of appearance of phosphorylcholine (PC)-responsive B cells, which are known to be acquired in ontogeny about 2 weeks later than DNP-responsive B cells, was also analyzed. The data indicate that CNF, unlike IL-1, IL-2, and WEHI-3 culture supernatant, results in an earlier appearance of PC-responsive B cells. These results suggest that soluble factors may play a role in the generation of the B cell repertoire.

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