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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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Evidence for influenza A virus on macrophages within the PD brain.(A–C): Representative staining (blue) within the SNpc of the PD brain utilizing mAB anti-influenza A virus antibody (35–481) depicting staining within large globular cells consistent with a morphology of macrophages (arrows, A–C). Panel B depicts staining within a macrophage-like cell extending an apparent pseudopod engulfing a smaller circular structure containing neuromelanin with punctate blue labeling (arrowhead, B). (D): Representative anti-influenza A virus staining (blue) in a control case identifying a single macrophage-like cell (arrowheads) surrounding a particle of neuromelanin material (arrow). (E): Quantitative analysis of the number of influenza A-positive macrophages labeled in Ctl (n = 5) or PD cases (n = 4) indicated a significant increase in PD cases (±S.D., p-value = 0.01). (F–K): Double labeling immunofluorescence in the SNpc of representative PD cases utilizing mAB anti-CD206, a macrophage marker (green, F and I), along with anti-influenza A virus labeling (35–481) (red, G and J) with the overlap image (yellow/orange, H and K). Two types of labeling were observed: In rounder macrophages (F), influenza A staining was more localized and homogenous (G). In perivascular regions, macrophages were more elongated (I) and influenza A labeling was clearly punctated in appearance (J). (L–N): Identical to Panels F–K, except with the use of a different antibody marker to both macrophages (CD14, green, L) and influenza A virus (35–483, red, M), with overlap image shown in Panel N. Punctate labeling of influenza A virus was observed on the surface of macrophages. (O): Control experiment showing lack of influenza A blue labeling in the absence of primary antibody. (P): Representative staining in a PD case utilizing an antibody against influenza B, indicating a general lack of blue labeling with this antibody. (Q): Panel Q depicts quantitative analysis indicating that approximately 80% of anti-CD206-positive cells co-localized with the anti-influenza A virus antibody (p = 4.55×10−6). Brown structures (A, D, O, and P) represent neuromelanin (asterisks), typical of neurons in the SNpc. All scale bars are 10 µm.
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pone-0020495-g005: Evidence for influenza A virus on macrophages within the PD brain.(A–C): Representative staining (blue) within the SNpc of the PD brain utilizing mAB anti-influenza A virus antibody (35–481) depicting staining within large globular cells consistent with a morphology of macrophages (arrows, A–C). Panel B depicts staining within a macrophage-like cell extending an apparent pseudopod engulfing a smaller circular structure containing neuromelanin with punctate blue labeling (arrowhead, B). (D): Representative anti-influenza A virus staining (blue) in a control case identifying a single macrophage-like cell (arrowheads) surrounding a particle of neuromelanin material (arrow). (E): Quantitative analysis of the number of influenza A-positive macrophages labeled in Ctl (n = 5) or PD cases (n = 4) indicated a significant increase in PD cases (±S.D., p-value = 0.01). (F–K): Double labeling immunofluorescence in the SNpc of representative PD cases utilizing mAB anti-CD206, a macrophage marker (green, F and I), along with anti-influenza A virus labeling (35–481) (red, G and J) with the overlap image (yellow/orange, H and K). Two types of labeling were observed: In rounder macrophages (F), influenza A staining was more localized and homogenous (G). In perivascular regions, macrophages were more elongated (I) and influenza A labeling was clearly punctated in appearance (J). (L–N): Identical to Panels F–K, except with the use of a different antibody marker to both macrophages (CD14, green, L) and influenza A virus (35–483, red, M), with overlap image shown in Panel N. Punctate labeling of influenza A virus was observed on the surface of macrophages. (O): Control experiment showing lack of influenza A blue labeling in the absence of primary antibody. (P): Representative staining in a PD case utilizing an antibody against influenza B, indicating a general lack of blue labeling with this antibody. (Q): Panel Q depicts quantitative analysis indicating that approximately 80% of anti-CD206-positive cells co-localized with the anti-influenza A virus antibody (p = 4.55×10−6). Brown structures (A, D, O, and P) represent neuromelanin (asterisks), typical of neurons in the SNpc. All scale bars are 10 µm.

Mentions: Because T-lymphocytes are known to respond to viral antigens and due to the long-standing idea that the triggering mechanism in PD involves an infectious agent [6], we examined PD cases utilizing an influenza A virus antibody known to detect numerous strains of the virus (Prosci, catalogue #35-481). We observed labeling on apparent macrophages in both PD (Fig. 5A–C) and control cases (Fig. 5D), although there was a significant increase (p = 0.01) in labeled macrophages of PD cases (Fig. 5E). Evidence for influenza A labeling was observed in all five PD cases examined and staining was absent in the presence of secondary antibody only (Fig. 5O). In addition, we examined PD cases for the presence of influenza B viral proteins, a virus not linked to PD and more commonly found in children than the elderly [17]. We were unable to detect any influenza B labeling in PD cases (Fig. 5P), suggesting a lack of this infectious agent in the PD brain.


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

Rohn TT, Catlin LW - PLoS ONE (2011)

Evidence for influenza A virus on macrophages within the PD brain.(A–C): Representative staining (blue) within the SNpc of the PD brain utilizing mAB anti-influenza A virus antibody (35–481) depicting staining within large globular cells consistent with a morphology of macrophages (arrows, A–C). Panel B depicts staining within a macrophage-like cell extending an apparent pseudopod engulfing a smaller circular structure containing neuromelanin with punctate blue labeling (arrowhead, B). (D): Representative anti-influenza A virus staining (blue) in a control case identifying a single macrophage-like cell (arrowheads) surrounding a particle of neuromelanin material (arrow). (E): Quantitative analysis of the number of influenza A-positive macrophages labeled in Ctl (n = 5) or PD cases (n = 4) indicated a significant increase in PD cases (±S.D., p-value = 0.01). (F–K): Double labeling immunofluorescence in the SNpc of representative PD cases utilizing mAB anti-CD206, a macrophage marker (green, F and I), along with anti-influenza A virus labeling (35–481) (red, G and J) with the overlap image (yellow/orange, H and K). Two types of labeling were observed: In rounder macrophages (F), influenza A staining was more localized and homogenous (G). In perivascular regions, macrophages were more elongated (I) and influenza A labeling was clearly punctated in appearance (J). (L–N): Identical to Panels F–K, except with the use of a different antibody marker to both macrophages (CD14, green, L) and influenza A virus (35–483, red, M), with overlap image shown in Panel N. Punctate labeling of influenza A virus was observed on the surface of macrophages. (O): Control experiment showing lack of influenza A blue labeling in the absence of primary antibody. (P): Representative staining in a PD case utilizing an antibody against influenza B, indicating a general lack of blue labeling with this antibody. (Q): Panel Q depicts quantitative analysis indicating that approximately 80% of anti-CD206-positive cells co-localized with the anti-influenza A virus antibody (p = 4.55×10−6). Brown structures (A, D, O, and P) represent neuromelanin (asterisks), typical of neurons in the SNpc. All scale bars are 10 µm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3105060&req=5

pone-0020495-g005: Evidence for influenza A virus on macrophages within the PD brain.(A–C): Representative staining (blue) within the SNpc of the PD brain utilizing mAB anti-influenza A virus antibody (35–481) depicting staining within large globular cells consistent with a morphology of macrophages (arrows, A–C). Panel B depicts staining within a macrophage-like cell extending an apparent pseudopod engulfing a smaller circular structure containing neuromelanin with punctate blue labeling (arrowhead, B). (D): Representative anti-influenza A virus staining (blue) in a control case identifying a single macrophage-like cell (arrowheads) surrounding a particle of neuromelanin material (arrow). (E): Quantitative analysis of the number of influenza A-positive macrophages labeled in Ctl (n = 5) or PD cases (n = 4) indicated a significant increase in PD cases (±S.D., p-value = 0.01). (F–K): Double labeling immunofluorescence in the SNpc of representative PD cases utilizing mAB anti-CD206, a macrophage marker (green, F and I), along with anti-influenza A virus labeling (35–481) (red, G and J) with the overlap image (yellow/orange, H and K). Two types of labeling were observed: In rounder macrophages (F), influenza A staining was more localized and homogenous (G). In perivascular regions, macrophages were more elongated (I) and influenza A labeling was clearly punctated in appearance (J). (L–N): Identical to Panels F–K, except with the use of a different antibody marker to both macrophages (CD14, green, L) and influenza A virus (35–483, red, M), with overlap image shown in Panel N. Punctate labeling of influenza A virus was observed on the surface of macrophages. (O): Control experiment showing lack of influenza A blue labeling in the absence of primary antibody. (P): Representative staining in a PD case utilizing an antibody against influenza B, indicating a general lack of blue labeling with this antibody. (Q): Panel Q depicts quantitative analysis indicating that approximately 80% of anti-CD206-positive cells co-localized with the anti-influenza A virus antibody (p = 4.55×10−6). Brown structures (A, D, O, and P) represent neuromelanin (asterisks), typical of neurons in the SNpc. All scale bars are 10 µm.
Mentions: Because T-lymphocytes are known to respond to viral antigens and due to the long-standing idea that the triggering mechanism in PD involves an infectious agent [6], we examined PD cases utilizing an influenza A virus antibody known to detect numerous strains of the virus (Prosci, catalogue #35-481). We observed labeling on apparent macrophages in both PD (Fig. 5A–C) and control cases (Fig. 5D), although there was a significant increase (p = 0.01) in labeled macrophages of PD cases (Fig. 5E). Evidence for influenza A labeling was observed in all five PD cases examined and staining was absent in the presence of secondary antibody only (Fig. 5O). In addition, we examined PD cases for the presence of influenza B viral proteins, a virus not linked to PD and more commonly found in children than the elderly [17]. We were unable to detect any influenza B labeling in PD cases (Fig. 5P), suggesting a lack of this infectious agent in the PD brain.

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