ADMINISTRATION OF IL-12 DURING ONGOING IMMUNE-RESPONSES FAILS TO PERMANENTLY SUPPRESS AND CAN EVEN ENHANCE THE SYNTHESIS OF ANTIGEN-SPECIFIC IGE

The synthesis of antibodies of the IgE isotype in mice largely depends on IL-4, a cytokine that is released by T lymphocytes of the T(h)2 subtype. IL-12 is a cytokine considered to direct Th cell development into a T(h)1 direction and to suppress T(h)2 responses including the synthesis of IgE. Here we report about the influence of IL-12 on the IgE response of mice immunized with protein antigens adsorbed to aluminum hydroxide. To avoid problems with the detection of IgE caused by an excess of competitive IgG antibodies produced in IL-12-treated mice, serum IgE was first extracted from the serum by plate-bound anti-IgE mAb and then determined either as total IgE or as antigen-specific IgE by using biotinylated anti-IgE or biotinylated antigen. Depending on the strain of mice and the dose of IL-12 injected together with the antigen, IL-12 can either temporarily suppress or augment the synthesis of (antigen-specific) IgE antibodies. This applies for CBA/J mice immunized six times in biweekly intervals with minute (0.1 mu g/injection) or three-times with large (5 mu g/injection) amounts of the bee venom allergen phospholipase A(2) (PLA(2)). Under both conditions the antibody response is characterized by the production of predominantly IgG1 as well as IgE but very little IgG2a, IgG2b and IgG3 antibodies. Simultaneous application of low doses of IL-12 (1 or 10 ng/day) led to a 2- to 4-fold enhancement of IgE production (PLA(2)-specific IgE or total IgE). Only a high dose of 1 mu g IL-12/day resulted in a 3- to 10-fold reduction of the IgE response. This suppression was not stable, however, because the synthesis of IgE antibodies was stimulated to a high level when these mice subsequently received a second course of immunizations in the absence of IL-12. Likewise, the synthesis of IgE was only temporarily suppressed by IL-12 treatment in CBA/J mice immunized with keyhole limpet hemocyanin (KLH) as antigen. However, application of low (10 ng/day) or high (1 mu g/day) doses of IL-12 during the primary course of immunizations of CBA/J mice with KLH suppressed the IgE response slightly or strongly respectively, in striking contrast, the KLH-specific IgE response of BALB/c mice was upregulated even when high doses of IL-12 (1 mu g/day) were injected simultaneously with the immunizations. Thus, these results demonstrate a great variability regarding the influence of IL-12 treatment on ongoing IgE responses in vivo.