ESSENTIAL ROLE OF TUMOR-NECROSIS-FACTOR-ALPHA IN THE DIFFERENTIATION OF HUMAN TONSIL INVIVO INDUCED B-CELLS CAPABLE OF SPONTANEOUS AND HIGH-RATE IMMUNOGLOBULIN SECRETION

Human tonsils contain B cells capable of spontaneous and high-rate immunoglobulin (Ig) secretion in vitro. These cells are in vivo induced mature B cells, and, as such, they provide an adequate model for studying tonsil B cell differentiation. The present report analyzes the effect of a variety of factors on purified tonsil B cells capable of spontaneous IgG secretion in fetal calf serum (FCS)-containing and serum-free supplemented cultures. Tumor necrosis factor-(TNF)alpha was found to be important for these B cells to reach the high-rate IgG-secreting stage, as is indicated by the following findings: (a) none of the factors used modified tonsil B cell IgG secretion in FCS-containing cultures; (b) TNF-alpha (5-20 ng/ml), but not other cytokines or factors including interleukin (IL)-6, was capable of restoring missing IgG production in serum-free supplemented cultures of tonsil B cells; and (c) IgG secretion in FCS-containing cultures was inhibited by the addition of blocking anti-TNF-alpha antibodies, but not anti-IL-6 antibodies, and this inhibition could be specifically reversed by exogenous TNF-alpha. TNF-alpha was actively produced by tonsil B cells (range 120-750 pg/ml) in the presence, but not in the absence, of FCS. The TNF-alpha inductive effect occurred during the first 12 h of culture and did not require DNA synthesis. These results indicate that the early and endogenous generation of TNF-alpha seems to be essential for tonsil in vivo induced B cells to differentiate into the high-rate Ig-secreting stage.