An organ fragment culture model to study lymphocyte activation in human lymphoid tissue.

We have established and evaluated an organ fragment culture model for the study of human lymphocyte activation and differentiation. Small fragments of tonsillar tissue were cultured on Gelfoam for periods of up to 7 days. Monoclonal antibody in the medium was able to diffuse into the tissue, as demonstrated by subsequent detection of antibody-coated cells. Phytohaemagglutinin added to the culture medium caused activation of T and B cells, as indicated by changes in expression of a number of markers. Antibody against human IgM (added as a F(ab')2 fragment) together with IL-4 caused B cell activation, detectable by an increased expression of CD23 and other markers. Cell viability fell gradually in culture, but useful data could nevertheless be obtained from culture periods up to 7 days. The organ fragment culture provides a model for the study of T and B cell activation which maintains, at least in part, the intercellular interactions and the native microenvironment of lymphoid tissue.

Modulation of expression of the antigen identified by FMC7 upon human B-lymphocyte activation: evidence for differences between activation in vivo and in vitro.

The antigen detected by the monoclonal antibody (mAb) FMC7 is expressed by a proportion of B lymphocytes, and previous information, based on leukaemias, cell lines and some normal cell studies, suggested that FMC7 reacts with relatively differentiated B cells. In this study, it is shown that cells in the high-density 'resting' fraction of tonsil B lymphocytes have a lower expression of FMC7 than cells in the low-density 'activated' fraction. However, activation of 'resting' B cells with anti-immunoglobulin (anti-IgM) together with IL-4 resulted in a decrease in FMC7 reactivity, whilst recognized markers of activation showed up-regulation, as expected. Activated cells which were cultured for an extra 3 days to allow return to a 'resting' stage showed an increase in FMC7 expression. Culture of activated B cells with low molecular weight B-cell growth factor (LMW-BCGF), tumour necrosis factor (TNF) or interleukin-6 (IL-6), did not lead to further changes in FMC7 reactivity, but stimulation with phorbol ester had an effect similar to that of anti-IgM + IL-4. These results suggest that FMC7 is a marker of differentiation rather than activation, and identifies a relatively mature subfraction of the pool of functionally mature B cells. Consistent with this interpretation, FMC7 and surface IgD tended to be expressed by reciprocal B-cell subpopulations.