The online version of this article (doi:10.1186/1471-2172-15-23) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
SW helped design, collect and analyze the data presented in this paper. PS and SH helped design the different experiments and also with the analysis of the data. SW, PS and SH have been involved in the preparation of this manuscript and writing the revisions. All authors approve the final version of this manuscript.
An accurate method that can diagnose and predict lupus and its neuropsychiatric manifestations is essential since currently there are no reliable methods. Autoantibodies to a varied panel of antigens in the body are characteristic of lupus. In this study we investigated whether serum autoantibody binding patterns on random-sequence peptide microarrays (immunosignaturing) can be used for diagnosing and predicting the onset of lupus and its central nervous system (CNS) manifestations. We also tested the techniques for identifying potentially pathogenic autoantibodies in CNS-Lupus. We used the well-characterized MRL/lpr lupus animal model in two studies as a first step to develop and evaluate future studies in humans.
In study one we identified possible diagnostic peptides for both lupus and altered behavior in the forced swim test. When comparing the results of study one to that of study two (carried out in a similar manner), we further identified potential peptides that may be diagnostic and predictive of both lupus and altered behavior in the forced swim test. We also characterized five potentially pathogenic brain-reactive autoantibodies, as well as suggested possible brain targets.
These results indicate that immunosignaturing could predict and diagnose lupus and its CNS manifestations. It can also be used to characterize pathogenic autoantibodies, which may help to better understand the underlying mechanisms of CNS-Lupus.
Additional file 1: Figure S2: Study 1- Behavioral Dysfunction (Forced Swim Test). A significant difference in float time was detected (F = 12.068, p < 0.008) and post-hoc analysis at p < 0.007 revealed that the 4 month MRL/lpr had significantly greater float times compared to the MRL/mp and C3H/HeJ. (PPT 142 KB)
Additional file 2: Figure S3: Study 1 – Group Separation within MRL/lpr by Anti-DNA Autoantibody Levels. The 4 M MRL/lpr mice were also split based on their anti-DNA antibody levels (grouping was similar to Figure 3). There was a significant difference between the groups (F = 91.176, p < 0.001). Utilizing post-hoc analysis at p < 0.004 there was a significant difference between the 4 month MRL/lpr with greater anti-DNA autoantibody levels and the 4 month MRL/lpr with lower anti-DNA autoantibody levels, the MRL/mp and the C3H/HeJ. There was also a significant difference between the MRL/lpr with lower anti-DNA autoantibody levels and the MRL/mp and the C3H/HeJ. (PPT 148 KB)
Additional file 3: Figure S4: Study 2 - Behavioral Dysfunction (Forced Swim Test). There was an overall significant difference between the groups (F = 11.057, p < 0.001) and post-hoc analysis at p < 0.05 revealed that the 4 M MRL/lpr floated significantly longer than the 1.5 M MRL/lpr, 1.5 M MRL/mp and 4 M MRL/mp. The 1.5 M MRL/mp was significantly different from the 4 M MRL/mp. (PPT 122 KB)
Additional file 4: Figure S1: Sample peptide binding intensities across pooled samples. This figure demonstrated the intensity pattern across individual mice of different strains. Each green dot is the binding of the serum to an individual peptide. (A) Secondary Only Control (only secondary and tertiary antibodies added). (B) C3H/HeJ. (C) MRL/lpr strain. (D) MRL/mp. (PPT 300 KB)
Additional file 5: Figure S5: Immunohistochemistry Control Slides. The above (orange-yellow) fluorescence is from propidium iodide binding to the cell nuclei. (A), (B), (C) and (D) showed that there was no binding in most of the brain section, the hippocampus, cortex and amygdala for the secondary only control. (E), (F), (G) and (H) showed that there was no binding in the whole brain, hippocampus, cortex and amygdala for the auto-fluorescence control. (PPT 470 KB)
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- Diagnosis and early detection of CNS-SLE in MRL/lpr mice using peptide microarrays
Steven A Hoffman
- BioMed Central
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