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. 2015 Feb 2;10(2):e0117935.
doi: 10.1371/journal.pone.0117935. eCollection 2015.

Nasal associated lymphoid tissue of the Syrian golden hamster expresses high levels of PrPC

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Nasal associated lymphoid tissue of the Syrian golden hamster expresses high levels of PrPC

Melissa D Clouse et al. PLoS One. .

Abstract

The key event in the pathogenesis of the transmissible spongiform encephalopathies is a template-dependent misfolding event where an infectious isoform of the prion protein (PrPSc) comes into contact with native prion protein (PrPC) and changes its conformation to PrPSc. In many extraneurally inoculated models of prion disease this PrPC misfolding event occurs in lymphoid tissues prior to neuroinvasion. The primary objective of this study was to compare levels of total PrPC in hamster lymphoid tissues involved in the early pathogenesis of prion disease. Lymphoid tissues were collected from golden Syrian hamsters and Western blot analysis was performed to quantify PrPC levels. PrPC immunohistochemistry (IHC) of paraffin embedded tissue sections was performed to identify PrPC distribution in tissues of the lymphoreticular system. Nasal associated lymphoid tissue contained the highest amount of total PrPC followed by Peyer's patches, mesenteric and submandibular lymph nodes, and spleen. The relative levels of PrPC expression in IHC processed tissue correlated strongly with the Western blot data, with high levels of PrPC corresponding with a higher percentage of PrPC positive B cell follicles. High levels of PrPC in lymphoid tissues closely associated with the nasal cavity could contribute to the relative increased efficiency of the nasal route of entry of prions, compared to other routes of infection.

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Conflict of interest statement

Competing Interests: Anthony E. Kincaid and Jason C. Bartz serve as Academic Editors for PLOS ONE, and thus are members of the PLOS ONE Editorial Board. The authors do not feel that this alters their adherence to PLOS ONE editorial policies and/or criteria.

Figures

Fig 1
Fig 1. NALT contains significantly more PrPC than other lymphoid tissues.
A) Western blot analysis and B) normalized quantification of lymphatic tissue PrPC abundance. SP—spleen, SLN—submandibular lymph node, MLN—mesenteric lymph node, PP—Peyer’s patch, NALT—nasal associated lymphoid tissue.
Fig 2
Fig 2. PrPC migration patterns were distinct for different lymphoid tissues.
A) Migration banding patterns are consistent when comparing individual animal homogenates. SP, SLN and PP homogenates from individual animals migrate similarly. B) Line graph of lane intensity analysis demonstrating peak differences in intensity of PrPC migration. C) Western blot of lymphoid tissue (NALT, SP, and PP) and BR control. D) Line graph of lane analysis of PrPC intensity of lymphoid tissue indicates intensity difference and migration profile of NALT. E) Comparison of PNGase treated and untreated PrPC from lymphoid tissue (SLN, PP and NALT and BR) demonstrates variable levels of full length and truncated PrPC in samples. BR—brain, SP—spleen, PP—Peyer’s patch, SLN—submandibular lymph node, NALT—nasal associated lymphoid tissue.
Fig 3
Fig 3. Immunohistochemistry (IHC) of lymphoid tissues demonstrating presence and localization of PrPC within B cell follicles.
IHC was performed on A/B) NALT, C) MLN, D) SLN, E) PP and F) SP with the anti-PrP antibody 3F4 (B-F) or an isotype control (A). The tissue sections were processed identically using the same reagents at the same time to illustrate relative differences in PrPC expression between tissues. The scale bar represents 100 μm. NALT—nasal associated lymphoid tissue, MLN—mesenteric lymph node, SLN—submandibular lymph node, PP—Peyer’s patch, SP—spleen.

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