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Immunolocalization of influenza A virus and markers of inflammation in the human Parkinson's disease brain.

Rohn TT, Catlin LW - PLoS ONE (2011)

Bottom Line: Although much is known regarding the molecular mechanisms leading to neuronal cell loss in Parkinson's disease (PD), the initiating event has not been identified.Prevailing theories including a chemical insult or infectious agent have been postulated as possible triggers, leading to neuroinflammation.Further supporting a role for neuroinflammation in PD was the identification of T-lymphocytes that colocalized with an antibody to caspase-cleaved Beclin-1 within the SNpc.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Boise State University, Boise, Idaho, United States of America. trohn@boisestate.edu

ABSTRACT
Although much is known regarding the molecular mechanisms leading to neuronal cell loss in Parkinson's disease (PD), the initiating event has not been identified. Prevailing theories including a chemical insult or infectious agent have been postulated as possible triggers, leading to neuroinflammation. We present immunohistochemical data indicating the presence of influenza A virus within the substantia nigra pars compacta (SNpc) from postmortem PD brain sections. Influenza A virus labeling was identified within neuromelanin granules as well as on tissue macrophages in the SNpc. Further supporting a role for neuroinflammation in PD was the identification of T-lymphocytes that colocalized with an antibody to caspase-cleaved Beclin-1 within the SNpc. The presence of influenza A virus together with macrophages and T-lymphocytes may contribute to the neuroinflammation associated with this disease.

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Caspase-cleaved Beclin-1 within T-lymphocytes in the PD brain.(A and B): Bright-field double labeling in the SNpc of a representative PD case (A) or control (B) illustrating co-localization of the BeclinCCP antibody (blue) with mAB CD3, a marker for T-lymphocytes (brown). Note the size and co-localization of both markers in cells (arrowheads, A), and in addition the single labeling of BeclinCCP within an apparent Lewy body (arrow, A). There was a general lack of labeling of both markers in control cases (B). Brown structures (A, B, F, and G) represent neuromelanin (asterisks), typical of neurons in the SNpc. (C–E): Representative immunofluorescence double labeling employing the BeclinCCP antibody (red, C) and mAB CD3 (green, D) in the SNpc of a representative PD case, with the overlapped image for both markers (yellow, E). (F and G): Representative staining in the SNpc of a PD case with anti-CD4 in (F) and anti-CD8 (G), markers for helper T-lymphocytes and cytotoxic T-lymphocytes, respectively (arrows). (H): Panel H depicts quantitative analysis indicating that approximately 78% of CD3-positive cells co-localized with the BeclinCCP antibody (p = 7.08×10−5). Scale bars represent 10 µm.
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pone-0020495-g004: Caspase-cleaved Beclin-1 within T-lymphocytes in the PD brain.(A and B): Bright-field double labeling in the SNpc of a representative PD case (A) or control (B) illustrating co-localization of the BeclinCCP antibody (blue) with mAB CD3, a marker for T-lymphocytes (brown). Note the size and co-localization of both markers in cells (arrowheads, A), and in addition the single labeling of BeclinCCP within an apparent Lewy body (arrow, A). There was a general lack of labeling of both markers in control cases (B). Brown structures (A, B, F, and G) represent neuromelanin (asterisks), typical of neurons in the SNpc. (C–E): Representative immunofluorescence double labeling employing the BeclinCCP antibody (red, C) and mAB CD3 (green, D) in the SNpc of a representative PD case, with the overlapped image for both markers (yellow, E). (F and G): Representative staining in the SNpc of a PD case with anti-CD4 in (F) and anti-CD8 (G), markers for helper T-lymphocytes and cytotoxic T-lymphocytes, respectively (arrows). (H): Panel H depicts quantitative analysis indicating that approximately 78% of CD3-positive cells co-localized with the BeclinCCP antibody (p = 7.08×10−5). Scale bars represent 10 µm.

Mentions: Single labeling experiments with the anti-oligodendrocyte antibody revealed little labeling of oligodendrocytes within the SNpc (Figure S2). Therefore, we hypothesized that an additional cell type was being labeled with the BeclinCCP antibody in this region. Based on the size and morphology of the cells labeled, we performed co-localization experiments with BeclinCCP and anti-CD3, a marker for T-lymphocytes. Co-localization of these two antibodies using bright field microscopy was evident within the SNpc in all PD cases examined (Fig. 4A), which was largely absent in age-matched control cases (Fig. 4B). Due to the lack of color separation using bright-field microscopy, additional co-localization experiments were undertaken using immunofluorescence (Fig. 4C–E). For immunofluorescence experiments, of those cells that were CD3-positive, 78% of these cells also were labeled with the BeclinCCP antibody (p = 7.08×10−5, ±S.E.M.) (Fig. 4H). The presence of caspase-cleaved Beclin-1 within T-lymphocytes would suggest caspase activation, a general feature of T-lymphocytes undergoing activation [12]. The presence of CD3-positive cells itself is not indicative of neuroinflammation, therefore, further experiments were performed to assess the type of T-lymphocytes using CD4 and CD8 antibodies. We detected the presence of both CD4+ and CD8+ T-lymphocytes in PD cases, and the staining profile was similar for both antibodies (Fig. 4F and G). In addition, staining of cells was often in areas of depigmentation (Fig. 4G), which may be indicative of neuroinflammation [13]. These results suggest the presence of both helper and cytotoxic T-lymphocytes in the SNpc of the PD brain, supporting previous studies [14], [15], [16].


Immunolocalization of influenza A virus and markers of inflammation in the human Parkinson's disease brain.

Rohn TT, Catlin LW - PLoS ONE (2011)

Caspase-cleaved Beclin-1 within T-lymphocytes in the PD brain.(A and B): Bright-field double labeling in the SNpc of a representative PD case (A) or control (B) illustrating co-localization of the BeclinCCP antibody (blue) with mAB CD3, a marker for T-lymphocytes (brown). Note the size and co-localization of both markers in cells (arrowheads, A), and in addition the single labeling of BeclinCCP within an apparent Lewy body (arrow, A). There was a general lack of labeling of both markers in control cases (B). Brown structures (A, B, F, and G) represent neuromelanin (asterisks), typical of neurons in the SNpc. (C–E): Representative immunofluorescence double labeling employing the BeclinCCP antibody (red, C) and mAB CD3 (green, D) in the SNpc of a representative PD case, with the overlapped image for both markers (yellow, E). (F and G): Representative staining in the SNpc of a PD case with anti-CD4 in (F) and anti-CD8 (G), markers for helper T-lymphocytes and cytotoxic T-lymphocytes, respectively (arrows). (H): Panel H depicts quantitative analysis indicating that approximately 78% of CD3-positive cells co-localized with the BeclinCCP antibody (p = 7.08×10−5). Scale bars represent 10 µm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3105060&req=5

pone-0020495-g004: Caspase-cleaved Beclin-1 within T-lymphocytes in the PD brain.(A and B): Bright-field double labeling in the SNpc of a representative PD case (A) or control (B) illustrating co-localization of the BeclinCCP antibody (blue) with mAB CD3, a marker for T-lymphocytes (brown). Note the size and co-localization of both markers in cells (arrowheads, A), and in addition the single labeling of BeclinCCP within an apparent Lewy body (arrow, A). There was a general lack of labeling of both markers in control cases (B). Brown structures (A, B, F, and G) represent neuromelanin (asterisks), typical of neurons in the SNpc. (C–E): Representative immunofluorescence double labeling employing the BeclinCCP antibody (red, C) and mAB CD3 (green, D) in the SNpc of a representative PD case, with the overlapped image for both markers (yellow, E). (F and G): Representative staining in the SNpc of a PD case with anti-CD4 in (F) and anti-CD8 (G), markers for helper T-lymphocytes and cytotoxic T-lymphocytes, respectively (arrows). (H): Panel H depicts quantitative analysis indicating that approximately 78% of CD3-positive cells co-localized with the BeclinCCP antibody (p = 7.08×10−5). Scale bars represent 10 µm.
Mentions: Single labeling experiments with the anti-oligodendrocyte antibody revealed little labeling of oligodendrocytes within the SNpc (Figure S2). Therefore, we hypothesized that an additional cell type was being labeled with the BeclinCCP antibody in this region. Based on the size and morphology of the cells labeled, we performed co-localization experiments with BeclinCCP and anti-CD3, a marker for T-lymphocytes. Co-localization of these two antibodies using bright field microscopy was evident within the SNpc in all PD cases examined (Fig. 4A), which was largely absent in age-matched control cases (Fig. 4B). Due to the lack of color separation using bright-field microscopy, additional co-localization experiments were undertaken using immunofluorescence (Fig. 4C–E). For immunofluorescence experiments, of those cells that were CD3-positive, 78% of these cells also were labeled with the BeclinCCP antibody (p = 7.08×10−5, ±S.E.M.) (Fig. 4H). The presence of caspase-cleaved Beclin-1 within T-lymphocytes would suggest caspase activation, a general feature of T-lymphocytes undergoing activation [12]. The presence of CD3-positive cells itself is not indicative of neuroinflammation, therefore, further experiments were performed to assess the type of T-lymphocytes using CD4 and CD8 antibodies. We detected the presence of both CD4+ and CD8+ T-lymphocytes in PD cases, and the staining profile was similar for both antibodies (Fig. 4F and G). In addition, staining of cells was often in areas of depigmentation (Fig. 4G), which may be indicative of neuroinflammation [13]. These results suggest the presence of both helper and cytotoxic T-lymphocytes in the SNpc of the PD brain, supporting previous studies [14], [15], [16].

Bottom Line: Although much is known regarding the molecular mechanisms leading to neuronal cell loss in Parkinson's disease (PD), the initiating event has not been identified.Prevailing theories including a chemical insult or infectious agent have been postulated as possible triggers, leading to neuroinflammation.Further supporting a role for neuroinflammation in PD was the identification of T-lymphocytes that colocalized with an antibody to caspase-cleaved Beclin-1 within the SNpc.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Boise State University, Boise, Idaho, United States of America. trohn@boisestate.edu

ABSTRACT
Although much is known regarding the molecular mechanisms leading to neuronal cell loss in Parkinson's disease (PD), the initiating event has not been identified. Prevailing theories including a chemical insult or infectious agent have been postulated as possible triggers, leading to neuroinflammation. We present immunohistochemical data indicating the presence of influenza A virus within the substantia nigra pars compacta (SNpc) from postmortem PD brain sections. Influenza A virus labeling was identified within neuromelanin granules as well as on tissue macrophages in the SNpc. Further supporting a role for neuroinflammation in PD was the identification of T-lymphocytes that colocalized with an antibody to caspase-cleaved Beclin-1 within the SNpc. The presence of influenza A virus together with macrophages and T-lymphocytes may contribute to the neuroinflammation associated with this disease.

Show MeSH
Related in: MedlinePlus