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*
Department of Veterinary Surgery, University of Osaka Prefecture College of Agriculture, Sakai, Osaka;
Noda Institute for Scientific Research, Noda, Noda-shi, Chiba-ken; and
Department of Veterinary Practice, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan;
§
College of Medicine, Jagiellonian University Krakow, Krakow, Poland; and
¶
Section of Allergy and Clinical Immunology, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520
Cutaneous painting with reactive haptens induces contact
sensitivity (CS) responses that are in vivo examples of T cell
immunity. In contrast, high dose i.v. administration of the hapten can
induce tolerance. We investigated the effect of IL-12 on reversal of
this tolerance and attempted to determine in vitro the mechanism of
this reversing effect by measuring proliferation and IFN-
production
by CS effector T cells stimulated with hapten-conjugated APC, and we
also measured CS ear swelling in vivo. The in vitro responses of T
cells to hapten-APC became absent in tolerized mice, paralleling
impaired in vivo CS responses. Addition of IL-12 to cultures
manifesting this fully established in vitro tolerance completely
restored impaired responses of tolerized T cells. The reversing effects
of IL-12 were not blocked by anti-IFN- mAb, but were blocked by
mAbs against B7-1, more strongly by anti-B7-2, and by both Abs
together. Additional in vivo ear-swelling response experiments
confirmed the reversing effects of IL-12 on established tolerance. To
examine whether the IL-12 effect depended on stimulation of IFN-, we
directly injected IFN- into tolerized mice. This partially mimicked
but did not fully reconstitute the effects of IL-12. In summary, IL-12
abrogation of established tolerance of CS may have been partially due
to endogenous production of IFN-, but appeared mainly due to direct
activation of the tolerized T cells by affecting signaling through
costimulatory molecules B7-1 and B7-2.
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