Autoantibodies to Collagen XVII Are Present in Parkinson’s Disease and Localize to Tyrosine-Hydroxylase Positive Neurons Kelly A.N. Messingham, Samantha.

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Autoantibodies to Collagen XVII Are Present in Parkinson’s Disease and Localize to Tyrosine-Hydroxylase Positive Neurons Kelly A.N. Messingham, Samantha Aust, Joseph Helfenberger, Krystal L. Parker, Susan Schultz, Julie McKillip, Nandakumar S. Narayanan, Janet A. Fairley Journal of Investigative Dermatology Volume 136, Issue 3, Pages 721-723 (March 2016) DOI: 10.1016/j.jid.2015.12.005 Copyright © 2015 The Authors Terms and Conditions

Figure 1 Serum reactivity to sec180 and NC16C regions of collagen XVII. Sera from 26 dementia and 24 Parkinson’s patients were screened for reactivity to recombinant sec180 or NC16C proteins by immunoblot. Reactivity was evaluated via densitometric analysis (NIH ImageJ) and values were normalized to the average density of negative control sera (assigned a value of 1, dotted line) and a 95% confidence interval (CI; dashed lines). (a) Sera from one dementia and seven Parkinson’s patients exhibited sec180 reactivity above the 95% CI and (b) sera from five dementia and two Parkinson’s patients exhibited reactivity to NC16C above the 95% CI. A single outlier (arrow) was identified using a Grubbs test (α = 0.01); omission of this outlier led to a significant (**P = 0.0027) difference in sec180 reactivity in dementia and Parkinson’s sera (Mann-Whitney test). No significant differences were observed in panel B. Journal of Investigative Dermatology 2016 136, 721-723DOI: (10.1016/j.jid.2015.12.005) Copyright © 2015 The Authors Terms and Conditions

Figure 2 Antibodies in Parkinson’s disease (PD) sera colocalize with dopaminergic neurons. Confocal images of cryosections of rat substantia nigra stained with anti-tyrosine hydroxylase (TH) antibodies (red) and PD sera (green). Using the colocalization finder tool in NIH Image J, areas of double staining in are pseudo-colored white; colocalization of IgG dopaminergic neurons (TH+) was observed with a blot-positive PD serum (a) but not a control PD serum (no binding of collagen XVII by blot) (b). To demonstrate the specificity of the co-localization, PD sera were pre-absorbed with gel-purified sec180 (-SEC180) or gel fragments devoid of protein (mock). The mock-absorption has no affect on co-localization of the signals (c) while preabsorption with sec180 results in a complete loss of co-localization (d). Images are representative of results obtained using sera from N = 5 individuals in each group. The scale bar = 100 μM. Journal of Investigative Dermatology 2016 136, 721-723DOI: (10.1016/j.jid.2015.12.005) Copyright © 2015 The Authors Terms and Conditions