24–26 It was reported that in MRL/lpr SLE-prone mice, deficiency of MIF resulted in attenuated glomerulonephritis and prolonged survival.27 Furthermore, elevated serum levels of MIF correlated with increased incidence of organ damage in patients with lupus.28 Therefore, in the present study, we analysed the selleck chemicals llc expression of the CD74/MIF pathway in (New Zealand Black × New Zealand White) F1 (BWF1) SLE-afflicted mice and investigated how treatment with the tolerogenic peptide, hCDR1, affected the CD74/MIF pathway. We demonstrate here the elevated expression of molecules of the CD74/MIF pathway on B cells and in two target organs, namely, brain

hippocampi and kidneys of SLE-afflicted mice. Treatment with hCDR1 down-regulated the expression of these molecules in association with up-regulation of B-cell apoptosis. PD-0332991 datasheet Female BWF1 mice were obtained from the Jackson Laboratory (Bar Harbor, ME). Mice were handled under protocols approved by the Weizmann Institute Animal Care and Use Committee according to international guidelines. The hCDR1,2 with sequence GYYWSWIRQPPGKGEEWIG, based on the CDR1 of a human monoclonal autoantibody,3 was synthesized by Polypeptide Laboratories (Torrance, CA). A peptide containing the same amino acids as hCDR1, with a scrambled order (SKGIPQYGGWPWEGWRYEI), designated scrambled peptide, was used as a control and PBS was used as a vehicle. Eight-month-old

BWF1 mice with established disease were divided into three groups (containing 8 to 12 mice) and injected subcutaneously with hCDR1, the scrambled control peptide (both 50 μg per mouse) or GABA Receptor vehicle alone, once a week for 10 weeks. Mice were followed for their clinical status [anti-double-stranded (ds) DNA autoantibodies and proteinuria] and at the end of treatment were killed and kidneys were analysed for the presence of immune complex deposits.4 Anti-dsDNA antibodies were detected using λ phage dsDNA, as previously described.4 Proteinuria was measured by a standard semi-quantitative test, using an Albustix kit (Bayer Diagnostic, Newbury, UK).

Detection of glomerular immune complex deposits was performed as described earlier.4 The intensity of immune complex deposits was graded as follows: 0, no immune complex deposits; 1, low intensity; 2, moderate intensity; and 3, high intensity of immune complexes. The immune complex deposit analysis was performed by two people blinded to whether the mice belonged to control or experimental groups. CD45R/B220+ cells were isolated from spleens of experimental mice using BD IMagnet (BD Biosciences, Chicago, IL), according to the manufacturer’s instructions. Briefly, splenocytes were suspended with CD45R/B220 particles, and incubated at 4° for 30 min. The cells labelled with IMag particles were placed in the BD IMagnet and were separated from unlabelled cells by magnetic force. The process was repeated once.