Dual role of interleukin-10 in murine Lyme disease: regulation of arthritis severity and host defense - PubMed (original) (raw)
Dual role of interleukin-10 in murine Lyme disease: regulation of arthritis severity and host defense
J P Brown et al. Infect Immun. 1999 Oct.
Abstract
In the murine model of Lyme disease, C3H/He mice exhibit severe arthritis while C57BL/6N mice exhibit mild lesions when infected with Borrelia burgdorferi. Joint tissues from these two strains of mice harbor similar concentrations of B. burgdorferi, suggesting that the difference in disease severity reflects differences in the magnitude of the inflammatory response to B. burgdorferi lipoproteins. Stimulation of bone marrow macrophages from C3H/HeN mice with the B. burgdorferi lipoprotein OspA resulted in higher-level production of the inflammatory mediators tumor necrosis factor alpha, nitric oxide, and interleukin-6 (IL-6) than that of macrophages from C57BL/6N mice. In contrast, macrophages from C57BL/6N mice consistently produced larger amounts of the anti-inflammatory cytokine IL-10 than did C3H/HeN macrophages. Addition of recombinant IL-10 suppressed the production of inflammatory mediators by macrophages from both strains. IL-10 was found to modulate B. burgdorferi-induced inflammation in vivo, since C57BL/6J mice deficient in IL-10 (IL-10-/-) developed more severe arthritis than wild-type C57BL/6J mice. The increase in arthritis severity was associated with a 10-fold decrease in the number of B. burgdorferi organisms present in ankle tissues from IL-10-/- mice. These findings suggest that in C57BL/6 mice, IL-10-dependent regulation of arthritis severity occurs at the expense of effective control of bacterial numbers.
Figures
FIG. 1
Production of inflammatory mediators by macrophages in response to OspA. Bone marrow-derived macrophages from C3H/HeN (circles) and C57BL/6N (squares) mice were stimulated with the indicated doses of OspA or LPS. Supernatants were assayed by ELISA (TNF-α and IL-6) or Griess assay (nitric oxide) at 6 h (TNF-α) or 24 h (nitric oxide and IL-6) after stimulation. Results for nitric oxide reflect supernatants in which agonists were added in the presence of 2 U of gamma interferon/ml. Data points represent duplicate samples, and results are representative of six experiments.
FIG. 2
Production of IL-10 by macrophages in response to OspA. Bone marrow-derived macrophages from C3H/HeN and C57BL/6N mice were stimulated with the indicated doses of OspA or LPS. Supernatants were assayed by ELISA at 24 h after stimulation. Data points represent duplicate samples, and results are representative of six experiments.
FIG. 3
Effects of exogenous IL-10 on production of inflammatory mediators in response to OspA. Bone marrow-derived macrophages from C3H/HeN and C57BL/6N mice were stimulated with the indicated doses of OspA in the presence of different concentrations of exogenous IL-10. Supernatants were assayed by ELISA at 6 h (TNF-α) or 24 h (IL-6) after stimulation. Data points represent duplicate samples, and results are representative of four experiments.
FIG. 4
Ankle swelling in different mouse strains infected with B. burgdorferi. The indicated mouse strains were infected by intradermal injection of 2,000 B. burgdorferi organisms, and ankles were measured weekly as described in Materials and Methods. Data points represent the averages and standard deviations of values for eight infected animals, and results are representative of two separate experiments. Mock-infected animals showed no increase in ankle swelling at any time point (data not shown).
FIG. 5
Lesion scores of joints from mice infected with B. burgdorferi. The indicated mouse strains were infected by intradermal injection of 2,000 B. burgdorferi organisms, and rear ankle joints were assessed for lesions. Each open circle represents the overall lesion score for an individual animal, and each black bar indicates the average score for the eight mice in a group. Uninfected controls exhibited normal histology (data not shown). These results are representative of two separate experiments.
FIG. 6
Detection of B. burgdorferi DNA in tissues from infected mice. Mice were infected by intradermal injection of 2,000 B. burgdorferi organisms, and the indicated tissues were harvested 4 weeks postinfection. Tissues were assessed for _B. burgdorferi_-specific DNA by continuously monitored PCR of the B. burgdorferi recA gene, and results were normalized to those of the host nidogen gene. Each data point represents the _B. burgdorferi_-specific DNA content of an individual infected tissue relative to nidogen, and each black bar indicates the average score for the eight mice in a group. Uninfected controls contained no _B. burgdorferi_-specific DNA (data not shown). These results are representative of two separate experiments.
FIG. 7
Detection of _B. burgdorferi_-specific Ig from infected mice. Serum was collected from the indicated mouse strains at 4 weeks after infection with 2,000 B. burgdorferi spirochetes. _B. burgdorferi_-specific Ig levels were determined by ELISA. Each data point represents the serum Ig content of an individual animal, and the black bars indicate the average scores. Uninfected controls contained no detectable _B. burgdorferi_-specific Ig (data not shown). These results are representative of two separate experiments.
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