IL-1Β-deficient mice are resistant to induction of experimental SLE

Illustrations

Figure 1 Serum levels of 16/6Id specific antibodies in IL-1-deficient mice. Control BALB/c mice and IL-1-deficient mice were immunized with the human monoclonal anti-DNA that bears the idiotype 16/6 Id (designated as 16/6Id). Anti-16/6Id antibodies were assessed by ELISA. Anti-16/6 antibody levels, 2 months after primary immunization with the 16/6Id are shown. The average values of O.D. of sera from 8-10 mice in each experimental point ± SD are shown.

Figure 2 Serum levels of anti-dsDNA antibodies in IL-1-deficient mice. Control BALB/c mice and IL-1-deficient mice were immunized with the 16/6Id antibody, which leads to the development of experimental SLE. Serum anti-dsDNA levels, 3 months after the primary immunization with the 16/6 Id antibody, are shown. Results are expressed as mean values of O.D. of sera from 8-10 mice at each experimental point ± SD, *p = 0.0043, **p = 0.007 as compared to the control group of BALB/c mice.

Figure 3 Proteinuria levels in IL-1-deficient mice following immunization with the 16/6/Id. Proteinuria was assessed, using a semiquantitative kit, as specified in Materials and Methods, in BALB/c control and IL-1-deficient mice, 7 months after the primary immunization with the 16/6 Id. Proteinuria levels (expressed as g/L) of 8-10 mice in each experimental group ± SD. *p = 0.004, **p = 0.025 as compared to the control group of BALB/c immunized mice are shown.

Figure 4 Immunohistology of kidney sections from IL-1-deficient mice following immunization with the 16/6Id. Frozen cryostat sections (5 μm) of kidneys of mice, 7 months following immunization with the 16/6Id were air-dried, fixed in acetone and stained with FITC-conjugated to goat antimouse IgG (γ chain specific) A- Control BALB/c mice; B- IL-1α^-/- mice; C- IL-1β^-/- mice; D- IL-1α/β^-/- mice; E- Normal (non-immunized) BALB/c mice. This figure demonstrates representative kidney sections from individual mice (x 400). Means of scores (see Materials and Methods) of immune complex deposits in kidney sections from 8-10 mice per group ± SD are indicated.P values for the significance between the 16/6Id immunized BALB/c mice and the IL-1^-/- relevant groups of mice are also shown.

Figure 5 Intracellular levels of cytokines in spleens from IL-1 deficient and BALB/c mice following immunization with the 16/6Id. Mice were sacrificed at the end of the experiment, 7 months after immunization with the 16/6 Id. Intracellular levels of cytokines in spleen cells were assessed by FACS analyses, using specific kits for assaying intracellular cytokines, as specified in the Materials and Methods. Pools of spleens from 8-10 mice per group were assessed for intracellular levels of cytokines. Each cytokine was measured in 3 samples. A representative staining from one experiment is shown.

Figure 6 Cytokine levels in supernatants of 16/6Id-stimulated spleen cells. Mice were sacrificed at the end of the experiment, 7 months after immunization with the 16/6 Id. Spleen cells were stimulated for 48 h in culture with the 16/6Id, and cytokine levels in supernatants were determined using ELISA kits, as specified in Materials and Methods. Pools of spleens from 8-10 mice per group were cultured in triplicate. Shown are mean values ± SD.*p < 0.03 as compared to the control group of BALB/c mice.