Paratuberculosis (PTB) or Johne's disease is a contagious enteritis of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Ovine PTB is less understood than bovine PTB, especially concerning paucibacillary infection and its evolution into clinical disease. We combined shotgun proteomics, histopathology and immunohistochemistry for the characterization of ileal tissues collected from seven asymptomatic sheep negative to serum ELISA, positive to feces and tissue MAP IS900 and F57 PCR, histologically classified as paucibacillary, actively infected, together with 3 MAP-free controls (K). Following shotgun proteomics with label-free quantitation and differential analysis, 96 proteins were significantly changed in PTB vs K, and were mostly involved in immune defense processes and in the macrophage-MAP interaction. Principal component analysis (PCA) of protein abundances highlighted two PTB sample clusters, PTB1 and PTB2, indicating a dichotomy in their proteomic profiles. This was in line with the PCA of histopathology data and was related to features of type 2 (PTB1) and type 3a (PTB2) lesions, respectively. PTB2 proteomes differed more than PTB1 proteomes from K: 43 proteins changed significantly only in PTB2 and 11 only in PTB1. The differential proteins cathelicidin, haptoglobin, S100A8 and S100A9 were evaluated by immunohistochemistry. K tissues were negative to cathelicidin and haptoglobin and sparsely positive to S100A8 and S100A9. PTB tissues were positive to all four proteins, with significantly more cells in PTB2 than in PTB1. In conclusion, we described several pathways altered in paucibacillary PTB, highlighted some proteomic differences among paucibacillary PTB cases, and identified potential markers for disease understanding, staging, and detection.

Characterization of paucibacillary ileal lesions in sheep with subclinical active infection by Mycobacterium avium subsp. Paratuberculosis / Pisanu, Salvatore; Cubeddu, Tiziana; Cacciotto, Carla; Pilicchi, Ylenia; Pagnozzi, Daniela; Uzzau, Sergio; Rocca, Stefano; Addis, Maria Filippa. - In: VETERINARY RESEARCH. - ISSN 0928-4249. - 49:1(2018). [10.1186/s13567-018-0612-0]

Characterization of paucibacillary ileal lesions in sheep with subclinical active infection by Mycobacterium avium subsp. Paratuberculosis

Pisanu, Salvatore;Cubeddu, Tiziana;Cacciotto, Carla;PILICCHI, YLENIA;Uzzau, Sergio;Rocca, Stefano;Addis, Maria Filippa
2018

Abstract

Paratuberculosis (PTB) or Johne's disease is a contagious enteritis of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Ovine PTB is less understood than bovine PTB, especially concerning paucibacillary infection and its evolution into clinical disease. We combined shotgun proteomics, histopathology and immunohistochemistry for the characterization of ileal tissues collected from seven asymptomatic sheep negative to serum ELISA, positive to feces and tissue MAP IS900 and F57 PCR, histologically classified as paucibacillary, actively infected, together with 3 MAP-free controls (K). Following shotgun proteomics with label-free quantitation and differential analysis, 96 proteins were significantly changed in PTB vs K, and were mostly involved in immune defense processes and in the macrophage-MAP interaction. Principal component analysis (PCA) of protein abundances highlighted two PTB sample clusters, PTB1 and PTB2, indicating a dichotomy in their proteomic profiles. This was in line with the PCA of histopathology data and was related to features of type 2 (PTB1) and type 3a (PTB2) lesions, respectively. PTB2 proteomes differed more than PTB1 proteomes from K: 43 proteins changed significantly only in PTB2 and 11 only in PTB1. The differential proteins cathelicidin, haptoglobin, S100A8 and S100A9 were evaluated by immunohistochemistry. K tissues were negative to cathelicidin and haptoglobin and sparsely positive to S100A8 and S100A9. PTB tissues were positive to all four proteins, with significantly more cells in PTB2 than in PTB1. In conclusion, we described several pathways altered in paucibacillary PTB, highlighted some proteomic differences among paucibacillary PTB cases, and identified potential markers for disease understanding, staging, and detection.
Characterization of paucibacillary ileal lesions in sheep with subclinical active infection by Mycobacterium avium subsp. Paratuberculosis / Pisanu, Salvatore; Cubeddu, Tiziana; Cacciotto, Carla; Pilicchi, Ylenia; Pagnozzi, Daniela; Uzzau, Sergio; Rocca, Stefano; Addis, Maria Filippa. - In: VETERINARY RESEARCH. - ISSN 0928-4249. - 49:1(2018). [10.1186/s13567-018-0612-0]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11388/219966
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