Limits...
Anti-Yo antibody uptake and interaction with its intracellular target antigen causes Purkinje cell death in rat cerebellar slice cultures: a possible mechanism for paraneoplastic cerebellar degeneration in humans with gynecological or breast cancers.

Greenlee JE, Clawson SA, Hill KE, Wood B, Clardy SL, Tsunoda I, Carlson NG - PLoS ONE (2015)

Bottom Line: However, intracellular accumulation of these antibodies did not affect Purkinje cell viability.The present study is the first to demonstrate that anti-Yo antibodies cause Purkinje cell death by binding to the intracellular 62 kDa Yo antigen.Purkinje cell death was not simply due to intraneuronal antibody accumulation.

View Article: PubMed Central - PubMed

Affiliation: Neurology Service, George E. Wahlen Veterans Affairs Health Care System, Salt Lake City, Utah, United States of America; Department of Neurology, University of Utah School of Medicine, Salt Lake City, Utah, United States of America.

ABSTRACT
Anti-Yo antibodies are immunoglobulin G (IgG) autoantibodies reactive with a 62 kDa Purkinje cell cytoplasmic protein. These antibodies are closely associated with paraneoplastic cerebellar degeneration in the setting of gynecological and breast malignancies. We have previously demonstrated that incubation of rat cerebellar slice cultures with patient sera and cerebrospinal fluid containing anti-Yo antibodies resulted in Purkinje cell death. The present study addressed three fundamental questions regarding the role of anti-Yo antibodies in disease pathogenesis: 1) Whether the Purkinje cell cytotoxicity required binding of anti-Yo antibody to its intraneuronal 62 kDa target antigen; 2) whether Purkinje cell death might be initiated by antibody-dependent cellular cytotoxicity rather than intracellular antibody binding; and 3) whether Purkinje cell death might simply be a more general result of intracellular antibody accumulation, rather than of specific antibody-antigen interaction. In our study, incubation of rat cerebellar slice cultures with anti-Yo IgG resulted in intracellular antibody binding, and cell death. Infiltration of the Purkinje cell layer by cells of macrophage/microglia lineage was not observed until extensive cell death was already present. Adsorption of anti-Yo IgG with its 62 kDa target antigen abolished both antibody accumulation and cytotoxicity. Antibodies to other intracellular Purkinje cell proteins were also taken up by Purkinje cells and accumulated intracellularly; these included calbindin, calmodulin, PCP-2, and patient anti-Purkinje cell antibodies not reactive with the 62 kDa Yo antigen. However, intracellular accumulation of these antibodies did not affect Purkinje cell viability. The present study is the first to demonstrate that anti-Yo antibodies cause Purkinje cell death by binding to the intracellular 62 kDa Yo antigen. Anti-Yo antibody cytotoxicity did not involve other antibodies or factors present in patient serum and was not initiated by brain mononuclear cells. Purkinje cell death was not simply due to intraneuronal antibody accumulation.

Show MeSH

Related in: MedlinePlus

Macrophage/microglial cells infiltrate the Purkinje cell layer after cell death has already begun.The Purkinje cell layer is shown at three points in time. Cells of macrophage/monocyte lineage were not detected within the Purkinje cell layer at 48 hours, or at 72 hours, when cell death was already apparent. Macrophage/mononuclear infiltration did not begin until 96 hours (data not shown) and was more extensive at 120 hours. These data indicate that initiation of Purkinje cell death did not involve antibody-mediated cellular cytotoxicity. Scale bar = 20μ.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4401511&req=5

pone.0123446.g003: Macrophage/microglial cells infiltrate the Purkinje cell layer after cell death has already begun.The Purkinje cell layer is shown at three points in time. Cells of macrophage/monocyte lineage were not detected within the Purkinje cell layer at 48 hours, or at 72 hours, when cell death was already apparent. Macrophage/mononuclear infiltration did not begin until 96 hours (data not shown) and was more extensive at 120 hours. These data indicate that initiation of Purkinje cell death did not involve antibody-mediated cellular cytotoxicity. Scale bar = 20μ.

Mentions: To determine whether Purkinje cell death might be due to interaction with brain mononuclear cells, cultures were incubated with anti-Yo antiserum, harvested at intervals between 48 and 120 hours, and co-labeled postfixation with anti-CD11b antibodies to detect macrophages/microglia. CD11b-positive cells were absent from the Purkinje cell layer at 48 hours and were still absent at 72 hours, when cell death was already widespread (Fig 3). Macrophage/microglial infiltrates became increasingly numerous as cell death became more extensive (Fig 3). These data indicate that Purkinje cell death preceded mononuclear cell infiltration, making it unlikely that brain mononuclear cells initiated Purkinje cell injury.


Anti-Yo antibody uptake and interaction with its intracellular target antigen causes Purkinje cell death in rat cerebellar slice cultures: a possible mechanism for paraneoplastic cerebellar degeneration in humans with gynecological or breast cancers.

Greenlee JE, Clawson SA, Hill KE, Wood B, Clardy SL, Tsunoda I, Carlson NG - PLoS ONE (2015)

Macrophage/microglial cells infiltrate the Purkinje cell layer after cell death has already begun.The Purkinje cell layer is shown at three points in time. Cells of macrophage/monocyte lineage were not detected within the Purkinje cell layer at 48 hours, or at 72 hours, when cell death was already apparent. Macrophage/mononuclear infiltration did not begin until 96 hours (data not shown) and was more extensive at 120 hours. These data indicate that initiation of Purkinje cell death did not involve antibody-mediated cellular cytotoxicity. Scale bar = 20μ.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401511&req=5

pone.0123446.g003: Macrophage/microglial cells infiltrate the Purkinje cell layer after cell death has already begun.The Purkinje cell layer is shown at three points in time. Cells of macrophage/monocyte lineage were not detected within the Purkinje cell layer at 48 hours, or at 72 hours, when cell death was already apparent. Macrophage/mononuclear infiltration did not begin until 96 hours (data not shown) and was more extensive at 120 hours. These data indicate that initiation of Purkinje cell death did not involve antibody-mediated cellular cytotoxicity. Scale bar = 20μ.
Mentions: To determine whether Purkinje cell death might be due to interaction with brain mononuclear cells, cultures were incubated with anti-Yo antiserum, harvested at intervals between 48 and 120 hours, and co-labeled postfixation with anti-CD11b antibodies to detect macrophages/microglia. CD11b-positive cells were absent from the Purkinje cell layer at 48 hours and were still absent at 72 hours, when cell death was already widespread (Fig 3). Macrophage/microglial infiltrates became increasingly numerous as cell death became more extensive (Fig 3). These data indicate that Purkinje cell death preceded mononuclear cell infiltration, making it unlikely that brain mononuclear cells initiated Purkinje cell injury.

Bottom Line: However, intracellular accumulation of these antibodies did not affect Purkinje cell viability.The present study is the first to demonstrate that anti-Yo antibodies cause Purkinje cell death by binding to the intracellular 62 kDa Yo antigen.Purkinje cell death was not simply due to intraneuronal antibody accumulation.

View Article: PubMed Central - PubMed

Affiliation: Neurology Service, George E. Wahlen Veterans Affairs Health Care System, Salt Lake City, Utah, United States of America; Department of Neurology, University of Utah School of Medicine, Salt Lake City, Utah, United States of America.

ABSTRACT
Anti-Yo antibodies are immunoglobulin G (IgG) autoantibodies reactive with a 62 kDa Purkinje cell cytoplasmic protein. These antibodies are closely associated with paraneoplastic cerebellar degeneration in the setting of gynecological and breast malignancies. We have previously demonstrated that incubation of rat cerebellar slice cultures with patient sera and cerebrospinal fluid containing anti-Yo antibodies resulted in Purkinje cell death. The present study addressed three fundamental questions regarding the role of anti-Yo antibodies in disease pathogenesis: 1) Whether the Purkinje cell cytotoxicity required binding of anti-Yo antibody to its intraneuronal 62 kDa target antigen; 2) whether Purkinje cell death might be initiated by antibody-dependent cellular cytotoxicity rather than intracellular antibody binding; and 3) whether Purkinje cell death might simply be a more general result of intracellular antibody accumulation, rather than of specific antibody-antigen interaction. In our study, incubation of rat cerebellar slice cultures with anti-Yo IgG resulted in intracellular antibody binding, and cell death. Infiltration of the Purkinje cell layer by cells of macrophage/microglia lineage was not observed until extensive cell death was already present. Adsorption of anti-Yo IgG with its 62 kDa target antigen abolished both antibody accumulation and cytotoxicity. Antibodies to other intracellular Purkinje cell proteins were also taken up by Purkinje cells and accumulated intracellularly; these included calbindin, calmodulin, PCP-2, and patient anti-Purkinje cell antibodies not reactive with the 62 kDa Yo antigen. However, intracellular accumulation of these antibodies did not affect Purkinje cell viability. The present study is the first to demonstrate that anti-Yo antibodies cause Purkinje cell death by binding to the intracellular 62 kDa Yo antigen. Anti-Yo antibody cytotoxicity did not involve other antibodies or factors present in patient serum and was not initiated by brain mononuclear cells. Purkinje cell death was not simply due to intraneuronal antibody accumulation.

Show MeSH
Related in: MedlinePlus