We found out an early manifestation of Th1-derived IL-2 and IFN-that vanished rapidly. IFN-that vanished rapidly. The second option might be due to the massive increase of IL-4 [21], which inhibits Th1 cells [22]. Our objective with the present study was to examine if a strong deviation towards Th1 would aggravate the autoimmune disease manifestations in HgIA. We used a treatment which has proved to be very efficient for ameliorating progressive cutaneous leishmaniasis in mice [23], a disease condition which shares with HgIA the dominance of Th2-connected cytokines [21]. We found that Th1 deviation accelerated several aspects of the HgIA condition. MATERIALS AND METHODS Animals Female A.SW (H-2s) mice were from M&B A/S (Ry, Denmark). The mice were housed in steel-wire cages under 12-h dark/12-h light cycles and given type R36 pellets (Lactamin, Vadstena, Sweden) and drinking water in the drinking water. Treatment with rIL-12 was started 18 h prior to Hg treatment and consisted of daily intraperitoneal (i.p.) injections of 02 < 005, Hg controls d< 005, Hg + rIL-12 e< 001, Hg settings fHg + rIL-12 g< 005, Hg + rIL-12 + -IL-4. In addition, in the pilot study, groups were given 1 mg mice STAT2 were used as positive settings and from normal mice as bad settings. Serum Ig isotype concentrations For analysis of serum IgM [26] microtitre plates were coated with rat <005 was regarded as statistically significant. RESULTS Animal health The animals showed no indications of disease during the experiment. A few mice died during or immediately after the blood samplings. Among the Hg settings two mice died at the blood samplings after 1 and 2 weeks. Two mice died in the group given Hg + rIL-12 + < 005; Fisher's exact test) and the titre (< 001) (Fig. 2a). The IgG-ANoA titre also continued to be increased significantly after 2 weeks treatment. rIL-12 is important for the acceleration of ANoA development because 71% of mice given Hg + rIL-12 but none of the mice given Hg + < 005, < 001 and 0001, respectively) compared with Hg settings as determined by KruskalCWallis test followed by Dunn's post-test. Effect of rIL-12 and/or -IL-4 treatment within the isotype pattern of Hg-induced ANoA The only significant effect of treatment with < 001) reduced compared to the Hg settings (Fig. 2)< 0001 and < 005, respectively), compared to the titre in the Hg settings. The same pattern was seen after 3 weeks, even though difference was significant (< 005) only in the group treated with rIL-12 + < 005) improved after 3 weeks in mice given either rIL-12 or < 005) (data not shown). Effect of rIL-12 and/or -IL-4 treatment within the Hg-induced polyclonal B-cell activation During the 3 weeks the Hg settings showed a reduction of the mean ideals for the PBA markers (IgM-< 005 Hg settings b< 001 Hg settings c< 0001 Hg settings d< 005 Hg + rIL-12 e< 001 Hg + rIL-12 + -IL-4. Effects of rIL-12 + -IL-4 treatment on Th1 and Th2 connected Ig isotypes in HgIA As evidence of Th2 suppression, after 1 week the Hg-treated mice receiving combined treatment with rIL-12 + < 005, NNC 55-0396 Fisher's precise test) fsignificantly different compared with Hg + rIL-12 (< 005, Fisher's precise test) gsignificantly different compared with Hg + rIL-12 (< 005; KruskalCWallis and Dunn's post-test) hsignificantly different compared with Hg + -IL-4 (< 001, Fisher's precise test) isignificantly different compared with Hg + -IL-4 (< 001, KruskalCWallis and Dunn's post-test) jsignificantly different compared with Hg settings (< 005, KruskalCWallis and Dunn's post-test) ksignificantly different compared with Hg settings (< 001, Fisher's precise test). Conversation The major objective of this study was to investigate if an autoimmune disease condition (HgIA) with characteristics of a Th2 type of reaction, but dependent on the Th1 cytokine IFN-[20], is definitely aggravated by deviating the immune response pattern NNC 55-0396 strongly from Th2 to Th1. We found that the Th1 deviation, accomplished by combined treatment with rIL-12 + [34,35] or [33,36] together with IL-4. With regard to additional HgIA parameters, treatment with rIL-12 improved the renal and splenic vessel wall IC deposits. The mechanism behind the development of vessel wall deposits in HgIA is not known, but the splenic deposits are more stable and less affected by manipulating HgIA compared NNC 55-0396 with the renal deposits [37,38]. This was also observed in the present NNC 55-0396 study, because treatment with secreting cells. While we did not measure IFN-secreting cells, a correlation is present between IFN-gene manifestation and the severity of HgIA [20]. Similarly, treatment of SJL (H-2s) mice with rIL-12 aggravates experimental sensitive encephalomyelitis (EAE) after adoptive transfer of antigen-stimulated lymph node cells [42]. However, in the EAE model the effect of.