Identification of a Serum-Induced Transcriptional Signature Associated With Type 1 Diabetes in the BioBreeding Rat

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FIG. 4.
FIG. 4.

Treatment of DRlyp/lyp rats with hIL-1Ra. A: Longitudinal monitoring of DRlyp/lyp rats treated with 350 μg/kg/day human recombinant IL-1Ra (n = 16, dashed line) or saline (n = 23, solid line). Agents were delivered intraperitoneally in saline. Treatment was initiated by day 30 (prior to insulitis). Fasting blood glucose was measured three times per week, and type 1 diabetes onset was defined as the first of two consecutive fasting blood glucose measurements >250 mg/dl. hIL-1Ra–treated rats survived 71 ± 11 days (range 53–100), while saline-treated controls survived 61 ± 6 days (53–75) (P = 0.007, log-rank test). B: Detection of anti–hIL-1Ra antibodies in IL-1Ra–treated DRlyp/lyp rats. Indicated amounts of hIL-1Ra (17 kDa) were loaded onto polyacrylamide gels, electrophoresed, and blotted. Membranes were probed with a 1:2,000 dilution of onset sera from hIL-1Ra–treated (top left blots), saline-treated (top right blot), day 40 hIL-1Ra–treated (bottom left blot), or saline-treated (bottom right blot) DRlyp/lyp rats. C: A Venn diagram illustrating the relationship between the gene expression induced between the PBS-treated DRlyp/lyp vs. BN allogeneic and hIL-1Ra–treated DRlyp/lyp vs. BN allogeneic inductions (244 |log2 ratio| >0.5- ±1.4-fold; FDR <0.10). PBMCs of six BN rats each were cultured with a serum pool generated from six hIL-1Ra–treated DRlyp/lyp rats or a serum pool generated from six PBS-treated DRlyp/lyp rats (n = 12 cultures). For PBMCs of each donor BN rat (n = 6), a culture possessing autologous sera was prepared. Fifteen cultures possessing allogeneic BN serum were prepared. Global gene expression was measured in each culture and all data were normalized with that of the autologous induction to account for gene expression induced by placing the PBMCs into culture. D: Regulated probes were identified between the PBS-treated DRlyp/lyp vs. BN allogeneic and hIL-1Ra–treated DRlyp/lyp vs. BN allogeneic inductions, replicates were averaged, and the relatedness of the three conditions were examined by hierarchical clustering. E: Well-annotated, regulated probe sets regulated by sera of PBS-treated DRlyp/lyp rats vs. the BN allogeneic induction. *Orthologues regulated by human type 1 diabetes sera (13). The scale represents the fold of change between the serum tested relative to autologous serum (−fourfold to +fourfold). (A high-quality digital representation of this figure is available in the online issue.)

This Article

  1. Diabetes vol. 59 no. 10 2375-2385