RESUMO
We report a rapidly progressive and fatal CD8 T-cell-mediated cerebellitis after ipilimumab (cytotoxic T-lymphocyte-associated protein 4 inhibitor) for small cell lung cancer. Clinical features and histopathology were consistent with an accelerated form of paraneoplastic cerebellar degeneration. A patchy CD8 T-cell infiltrate spatially corresponded to areas of Purkinje cell loss, with occasional CD8 polarisation towards Purkinje cells. CD20-positive B cells were sparse. CD8 T-cell-mediated cerebellitis after immune checkpoint inhibitor treatment may recapitulate the early stages of paraneoplastic cerebellar degeneration.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Inibidores de Checkpoint Imunológico/efeitos adversos , Ipilimumab/efeitos adversos , Neoplasias Pulmonares/tratamento farmacológico , Degeneração Paraneoplásica Cerebelar/induzido quimicamente , Células de Purkinje/imunologia , Carcinoma de Pequenas Células do Pulmão/tratamento farmacológico , Humanos , Inibidores de Checkpoint Imunológico/administração & dosagem , Inibidores de Checkpoint Imunológico/uso terapêutico , Ipilimumab/administração & dosagem , Ipilimumab/uso terapêutico , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Degeneração Paraneoplásica Cerebelar/imunologia , Células de Purkinje/efeitos dos fármacos , Carcinoma de Pequenas Células do Pulmão/imunologia , Carcinoma de Pequenas Células do Pulmão/patologiaRESUMO
OBJECTIVE: To describe the identification of regulator of G-protein signaling 8 (RGS8) as an autoantibody target in patients with cerebellar syndrome associated with lymphoma. METHODS: Sera of 4 patients with a very similar unclassified reactivity against cerebellar Purkinje cells were used in antigen identification experiments. Immunoprecipitations with cerebellar lysates followed by mass spectrometry identified the autoantigen, which was verified by recombinant immunofluorescence assay, immunoblot, and ELISA with the recombinant protein. RESULTS: The sera and CSF of 4 patients stained the Purkinje cells and molecular layer of the cerebellum. RGS8 was identified as the target antigen in all 4 sera. In a neutralization experiment, recombinant human RGS8 was able to neutralize the autoantibodies' tissue reaction. Patient sera and CSF showed a specific reactivity against recombinant RGS8 in ELISA and immunoblot, whereas no such reactivity was detectable in the controls. Clinical data were available for 2 of the 4 patients, remarkably both presented with cerebellar syndrome accompanied by B-cell lymphoma of the stomach (patient 1, 53 years) or Hodgkin lymphoma (patient 2, 74 years). CONCLUSION: Our results indicate that autoantibodies against the intracellular Purkinje cell protein RGS8 represent new markers for paraneoplastic cerebellar syndrome associated with lymphoma. CLASSIFICATION OF EVIDENCE: This study provided Class IV evidence that autoantibodies against the intracellular Purkinje cell protein RGS8 are associated with paraneoplastic cerebellar syndrome in lymphoma.
Assuntos
Autoanticorpos/imunologia , Degeneração Paraneoplásica Cerebelar/imunologia , Células de Purkinje/imunologia , Proteínas RGS/metabolismo , Idoso , Animais , Cerebelo/patologia , Feminino , Células HEK293 , Haplorrinos , Voluntários Saudáveis , Humanos , Linfoma/imunologia , Masculino , Pessoa de Meia-Idade , Degeneração Paraneoplásica Cerebelar/complicações , RatosRESUMO
OBJECTIVE: Identify the subcellular location and potential binding partners of two cerebellar degeneration-related proteins, CDR2L and CDR2, associated with anti-Yo-mediated paraneoplastic cerebellar degeneration. METHODS: Cancer cells, rat Purkinje neuron cultures, and human cerebellar sections were exposed to cerebrospinal fluid and serum from patients with paraneoplastic cerebellar degeneration with Yo antibodies and with several antibodies against CDR2L and CDR2. We used mass spectrometry-based proteomics, super-resolution microscopy, proximity ligation assay, and co-immunoprecipitation to verify the antibodies and to identify potential binding partners. RESULTS: We confirmed the CDR2L specificity of Yo antibodies by mass spectrometry-based proteomics and found that CDR2L localized to the cytoplasm and CDR2 to the nucleus. CDR2L co-localized with the 40S ribosomal protein S6, while CDR2 co-localized with the nuclear speckle proteins SON, eukaryotic initiation factor 4A-III, and serine/arginine-rich splicing factor 2. INTERPRETATION: We showed that Yo antibodies specifically bind to CDR2L in Purkinje neurons of PCD patients where they potentially interfere with the function of the ribosomal machinery resulting in disrupted mRNA translation and/or protein synthesis. Our findings demonstrating that CDR2L interacts with ribosomal proteins and CDR2 with nuclear speckle proteins is an important step toward understanding PCD pathogenesis.
Assuntos
Autoanticorpos , Autoantígenos , Núcleo Celular , Citoplasma , Proteínas do Tecido Nervoso , Degeneração Paraneoplásica Cerebelar , Biossíntese de Proteínas , Células de Purkinje , Animais , Autoanticorpos/imunologia , Autoanticorpos/metabolismo , Autoantígenos/imunologia , Autoantígenos/metabolismo , Linhagem Celular Tumoral , Núcleo Celular/imunologia , Núcleo Celular/metabolismo , Citoplasma/imunologia , Citoplasma/metabolismo , Humanos , Imunoprecipitação , Espectrometria de Massas , Proteínas do Tecido Nervoso/imunologia , Proteínas do Tecido Nervoso/metabolismo , Degeneração Paraneoplásica Cerebelar/imunologia , Degeneração Paraneoplásica Cerebelar/metabolismo , Biossíntese de Proteínas/fisiologia , Proteômica , Células de Purkinje/imunologia , Células de Purkinje/metabolismo , RatosRESUMO
Paraneoplastic cerebellar degeneration (PCD) is a rare immune-mediated disease that develops mostly in the setting of neoplasia and offers a unique prospect to explore the interplay between tumor immunity and autoimmunity. In PCD, the deleterious adaptive immune response targets self-antigens aberrantly expressed by tumor cells, mostly gynecological cancers, and physiologically expressed by the Purkinje neurons of the cerebellum. Highly specific anti-neuronal antibodies in the serum and cerebrospinal fluid represent key diagnostic biomarkers of PCD. Some anti-neuronal antibodies such as anti-Yo autoantibodies (recognizing the CDR2/CDR2L proteins) are only associated with PCD. Other anti-neuronal antibodies, such as anti-Hu, anti-Ri, and anti-Ma2, are detected in patients with PCD or other types of paraneoplastic neurological manifestations. Importantly, these autoantibodies cannot transfer disease and evidence for a pathogenic role of autoreactive T cells is accumulating. However, the precise mechanisms responsible for disruption of self-tolerance to neuronal self-antigens in the cancer setting and the pathways involved in pathogenesis within the cerebellum remain to be fully deciphered. Although the occurrence of PCD is rare, the risk for such severe complication may increase with wider use of cancer immunotherapy, notably immune checkpoint blockade. Here, we review recent literature pertaining to the pathophysiology of PCD and propose an immune scheme underlying this disabling disease. Additionally, based on observations from patients' samples and on the pre-clinical model we recently developed, we discuss potential therapeutic strategies that could blunt this cerebellum-specific autoimmune disease.
Assuntos
Antígenos de Neoplasias/imunologia , Autoanticorpos/imunologia , Autoimunidade , Cerebelo/imunologia , Neoplasias/imunologia , Degeneração Paraneoplásica Cerebelar/imunologia , Animais , Autoantígenos/imunologia , Autoantígenos/metabolismo , Cerebelo/metabolismo , Cerebelo/patologia , Humanos , Imunoterapia/efeitos adversos , Neoplasias/metabolismo , Neoplasias/patologia , Neoplasias/terapia , Proteínas do Tecido Nervoso/imunologia , Proteínas do Tecido Nervoso/metabolismo , Degeneração Paraneoplásica Cerebelar/metabolismo , Degeneração Paraneoplásica Cerebelar/patologia , Células de Purkinje/imunologia , Células de Purkinje/metabolismo , Células de Purkinje/patologia , Fatores de Risco , Linfócitos T/imunologia , Linfócitos T/metabolismoRESUMO
BACKGROUND: Patients with liver cirrhosis may develop hepatic encephalopathy. Rats with chronic hyperammonemia exhibit neurological alterations mediated by peripheral inflammation and neuroinflammation. Motor incoordination is due to increased TNF-a levels and activation of its receptor TNFR1 in the cerebellum. The aims were to assess (a) whether peripheral inflammation is responsible for TNF-a induction in hyperammonemic rats, (b) the cell type(s) in which TNF-a is increased, (c) whether this increase is associated with increased nuclear NF-κB and TNFR1 activation, (d) the time course of TNF-a induction, and (e) if TNF-a is induced in the Purkinje neurons of patients who die with liver cirrhosis. METHODS: We analyzed the level of TNF-a mRNA and NF-κB in microglia, astrocytes, and Purkinje neurons in the cerebellum after 1, 2, and 4 weeks of hyperammonemia. We assessed whether preventing peripheral inflammation by administering an anti-TNF-a antibody prevents TNF-a induction. We tested whether TNF-a induction is reversed by R7050, which inhibits the TNFR1-NF-κB pathway, in ex vivo cerebellar slices. RESULTS: Hyperammonemia induced microglial and astrocyte activation at 1 week. This was followed by TNF-a induction in both glial cell types at 2 weeks and in Purkinje neurons at 4 weeks. The level of TNF-a mRNA increased in parallel with the TNF-a protein level, indicating that TNF-a was synthesized in Purkinje cells. This increase was associated with increased NF-κB nuclear translocation. The nuclear translocation of NF-κB and the increase in TNF-a were reversed by R7050, indicating that they were mediated by the activation of TNFR1. Preventing peripheral inflammation with an anti-TNF-a antibody prevents TNF-a induction. CONCLUSION: Sustained (4 weeks) but not short-term hyperammonemia induces TNF-a in Purkinje neurons in rats. This is mediated by peripheral inflammation. TNF-a is also increased in the Purkinje neurons of patients who die with liver cirrhosis. The results suggest that hyperammonemia induces TNF-a in glial cells and that TNF-a released by glial cells activates TNFR1 in Purkinje neurons, leading to NF-κB nuclear translocation and the induction of TNF-a expression, which may contribute to the neurological alterations observed in hyperammonemia and hepatic encephalopathy.
Assuntos
Cerebelo/metabolismo , Hiperamonemia/metabolismo , Células de Purkinje/metabolismo , Transdução de Sinais/fisiologia , Fator de Necrose Tumoral alfa/metabolismo , Idoso , Animais , Cerebelo/imunologia , Humanos , Hiperamonemia/complicações , Hiperamonemia/imunologia , Cirrose Hepática/complicações , Cirrose Hepática/imunologia , Cirrose Hepática/metabolismo , Masculino , Pessoa de Meia-Idade , NF-kappa B/imunologia , NF-kappa B/metabolismo , Neuroglia/imunologia , Neuroglia/metabolismo , Células de Purkinje/imunologia , Ratos , Ratos Wistar , Receptores Tipo I de Fatores de Necrose Tumoral/imunologia , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Fator de Necrose Tumoral alfa/imunologiaAssuntos
Neoplasias das Tubas Uterinas/complicações , Degeneração Paraneoplásica Cerebelar/complicações , Idoso de 80 Anos ou mais , Autoanticorpos/análise , Neoplasias das Tubas Uterinas/diagnóstico , Feminino , Humanos , Degeneração Paraneoplásica Cerebelar/diagnóstico , Degeneração Paraneoplásica Cerebelar/imunologia , Células de Purkinje/imunologiaRESUMO
Paraneoplastic neurological disorders result from an autoimmune response against neural self-antigens that are ectopically expressed in neoplastic cells. In paraneoplastic disorders associated to autoantibodies against intracellular proteins, such as paraneoplastic cerebellar degeneration (PCD), current data point to a major role of cell-mediated immunity. In an animal model, in which a neo-self-antigen was expressed in both Purkinje neurons and implanted breast tumor cells, immune checkpoint blockade led to complete tumor control at the expense of cerebellum infiltration by T cells and Purkinje neuron loss, thereby mimicking PCD. Here, we identify 2 potential therapeutic targets expressed by cerebellum-infiltrating T cells in this model, namely α4 integrin and IFN-γ. Mice with PCD were treated with anti-α4 integrin antibodies or neutralizing anti-IFN-γ antibodies at the onset of neurological signs. Although blocking α4 integrin had little or no impact on disease development, treatment using the anti-IFN-γ antibody led to almost complete protection from PCD. These findings strongly suggest that the production of IFN-γ by cerebellum-invading T cells plays a major role in Purkinje neuron death. Our successful preclinical use of neutralizing anti-IFN-γ antibody for the treatment of PCD offers a potentially new therapeutic opportunity for cancer patients at the onset of paraneoplastic neurological disorders.
Assuntos
Interferon gama/antagonistas & inibidores , Neoplasias Mamárias Experimentais/complicações , Degeneração Paraneoplásica Cerebelar/tratamento farmacológico , Células de Purkinje/patologia , Linfócitos T/efeitos dos fármacos , Animais , Antígenos de Neoplasias/imunologia , Autoanticorpos/imunologia , Autoantígenos/imunologia , Linhagem Celular Tumoral/transplante , Feminino , Integrina alfa4/antagonistas & inibidores , Integrina alfa4/imunologia , Integrina alfa4/metabolismo , Interferon gama/imunologia , Interferon gama/metabolismo , Neoplasias Mamárias Experimentais/imunologia , Camundongos , Camundongos Knockout , Degeneração Paraneoplásica Cerebelar/imunologia , Degeneração Paraneoplásica Cerebelar/patologia , Células de Purkinje/imunologia , Células de Purkinje/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismoAssuntos
Linfócitos T CD8-Positivos/imunologia , Antígeno HLA-A2/imunologia , Imunoterapia Adotiva , Células de Purkinje/imunologia , Tirosina Quinase 3 Semelhante a fms/metabolismo , Animais , Linfócitos T CD8-Positivos/metabolismo , Células Cultivadas , Antígeno HLA-A2/metabolismo , Humanos , Células K562 , Camundongos , Células de Purkinje/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismoRESUMO
The paraneoplastic cerebellar syndrome presents as severe neuroimmunological disease associated with malignancies. Antibodies against antigens expressed by tumor cells cross-react with proteins of cerebellar Purkinje cells leading to neuroinflammation and neuronal loss. These antineuronal antibodies are preferentially investigated by serological analyses while examination of the cerebrospinal fluid is only performed infrequently. We retrospectively investigated 12 patients with antineuronal antibodies against Purkinje cells with a special focus on cerebrospinal fluid. Our results confirm a subacute disease with a severe cerebellar syndrome in 10 female patients due to anti-Yo antibodies associated mostly with gynecological malignancies. While standard cerebrospinal fluid parameters infrequently revealed pathological results, all patients presented oligoclonal bands indicating intrathecal IgG synthesis. Analyses of anti-Yo antibodies in cerebrospinal fluid by calculating the antibody specific index revealed intrathecal synthesis of anti-Yo antibodies in these patients. In analogy to anti-Yo syndrome, an intrathecal production of anti-Tr antibodies in one patient who presented with a paraneoplastic cerebellar syndrome was detected. In an additional patient, anti-Purkinje cell antibodies of unknown origin in the cerebrospinal fluid but not in serum were determined suggesting an isolated immune reaction within the central nervous system (CNS) and underlining the importance of investigating the cerebrospinal fluid. In conclusion, patients with a cerebellar syndrome display a distinct immune reaction within the cerebrospinal fluid including intrathecal synthesis of disease-specific antibodies. We emphasize the importance of a thorough immunological work up including investigations of both serum and cerebrospinal fluid.
Assuntos
Autoanticorpos/metabolismo , Proteínas do Tecido Nervoso/imunologia , Degeneração Paraneoplásica Cerebelar/imunologia , Degeneração Paraneoplásica Cerebelar/patologia , Células de Purkinje/metabolismo , Receptores de Superfície Celular/imunologia , Adulto , Idoso , Feminino , Humanos , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Degeneração Paraneoplásica Cerebelar/diagnóstico por imagem , Degeneração Paraneoplásica Cerebelar/metabolismo , Células de Purkinje/imunologia , Estudos RetrospectivosRESUMO
Antibodies against brain proteins were identified in the plasma of cancer patients and are defined to cause paraneoplastic neurological syndromes. The profiles of brain-directed antibodies in non-small cell lung cancer (NSCLC) are largely unknown. Here, for the first time, we compared autoantibodies against brain proteins in NSCLC (n = 18) against those present in age-matched non-cancer control subjects (n = 18) with a similar life-style, habit, and medical history. Self-recognizing immunoglobulin (IgG) are primarily directed against cells in the cortex (P = 0.008), hippocampus (P = 0.003-0.05), and cerebellum (P = 0.02). More specifically, IgG targets were prominent in the pyramidal, Purkinje, and granule cell layers. Furthermore, autoimmune IgG signals were localized to neurons (81%), astrocytes (48%), and endothelial (29%) cells. While cancer sera yielded overall higher intensity signals, autoantigens of 100, 65, 45, 37, and 30 kDa molecular weights were the most represented. Additionally, a group of 100 kDa proteins seem more prevalent in female adenocarcinoma patients (4/5, 80%). In conclusion, our results revealed autoantigen specificity in NSCLC, which implicitly depends on patient's demographics and disease history. Patients at risk for lung cancer but with no active disease revealed that the immune profile in NSCLC is disease-dependent.
Assuntos
Autoanticorpos/imunologia , Encéfalo/imunologia , Carcinoma Pulmonar de Células não Pequenas/imunologia , Neoplasias Pulmonares/imunologia , Adenocarcinoma/sangue , Adenocarcinoma/imunologia , Idoso , Idoso de 80 Anos ou mais , Animais , Antígenos Nucleares/imunologia , Astrócitos/imunologia , Autoanticorpos/sangue , Autoantígenos/química , Autoantígenos/imunologia , Western Blotting , Carcinoma Pulmonar de Células não Pequenas/sangue , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imuno-Histoquímica , Neoplasias Pulmonares/sangue , Masculino , Pessoa de Meia-Idade , Peso Molecular , Proteínas do Tecido Nervoso/imunologia , Neurônios/imunologia , Células de Purkinje/imunologia , Células Piramidais/imunologia , Ratos Sprague-DawleyRESUMO
68-year-old female presented with involuntary movements. MRI was normal. Cerebrospinal fluid analysis was normal. whole body CT and biopsy confirmed diagnosis of metastatic adenocarnimoa. The autoimmune panel was positive for anti-Yo antibodies.
Assuntos
Anticorpos/sangue , Coreia/imunologia , Discinesias/imunologia , Proteínas do Tecido Nervoso/imunologia , Síndromes Paraneoplásicas do Sistema Nervoso/imunologia , Células de Purkinje/imunologia , Idoso , Coreia/sangue , Discinesias/sangue , Feminino , Humanos , Imageamento por Ressonância Magnética , Síndromes Paraneoplásicas do Sistema Nervoso/sangueRESUMO
OBJECTIVE: To report the identification of microtubule-associated protein (MAP) 1B as the antigen of the previously described Purkinje cell cytoplasmic antibody type 2 (PCA-2) antibody, its frequency, and clinical, oncological, and serological associations. METHODS: Archival serum or cerebrospinal fluid (CSF) specimens were available from 96 of 118 consecutive PCA-2-IgG-seropositive patients identified during 1993-2016. The autoantigen, defined in mouse brain lysate by Western blot and mass spectrometry, was confirmed by dual immunohistochemical staining using commercial antibodies. The major antigenic region was defined by Western blot using recombinant protein fragments. RESULTS: IgG in 95 of 96 patients' serum or CSF (but in none of 98 healthy or disease control subjects' serum specimens) bound to recombinant MAP1B. A minority (17.5%) of patients' IgG also bound to MAP1A. PCA-2 was often accompanied by additional neural autoantibody markers of small-cell carcinoma, including collapsin response-mediated protein 5 (CRMP5) IgG (26%) or antineuronal nuclear antibody type 1 (ANNA-1) IgG (also known as anti-Hu; 13%). Neurological manifestations in 95 patients were (in decreasing frequency): peripheral neuropathy, 53%; cerebellar ataxia, dysmetria, or dysarthria, 38%; and encephalopathy, 27%. Cancer (majority small-cell lung carcinoma [SCLC]) was detected in 66 of 84 evaluated patients (79%). The MAP1B (PCA-2) autoantibody detection rate, among approximately 70,000 patients undergoing service neural autoantibody evaluation in 2015, was 0.024%, equaling amphiphysin IgG (0.026%) and more common than ANNA-2 (also known as anti-Ri; 0.016%) and PCA-Tr (also known as delta/notch-like epidermal growth factor-related receptor [DNER]; 0.006%). INTERPRETATION: MAP1B, the PCA-2 autoantigen, represents a novel target in paraneoplastic neurological disorders and has high predictive value for SCLC. Its relatively high prevalence, compared with other recognized paraneoplastic neural autoantibodies, justifies its testing in comprehensive paraneoplastic neural autoantibody evaluation. Ann Neurol 2017;81:266-277.
Assuntos
Autoanticorpos/imunologia , Autoantígenos/imunologia , Biomarcadores Tumorais/imunologia , Neoplasias Pulmonares/imunologia , Proteínas Associadas aos Microtúbulos/imunologia , Síndromes Paraneoplásicas do Sistema Nervoso/imunologia , Células de Purkinje/imunologia , Carcinoma de Pequenas Células do Pulmão/imunologia , Animais , Humanos , Imunoglobulina G/imunologia , Neoplasias Pulmonares/diagnóstico , Camundongos , Síndromes Paraneoplásicas do Sistema Nervoso/sangue , Síndromes Paraneoplásicas do Sistema Nervoso/líquido cefalorraquidiano , Proteínas Recombinantes , Carcinoma de Pequenas Células do Pulmão/diagnósticoRESUMO
Patients with Purkinje cell cytoplasmic autoantibody type 2 (PCA-2) and collapsin response-mediator protein-5 (CRMP-5) autoantibody can present with multifocal elements of encephalomyeloneuropathy. Except for an anecdotal report, case descriptions of paraneoplastic small fibre neuropathy are lacking. We report paraneoplastic small fibre neuropathy followed by chorea associated with small cell lung cancer. A man aged 57 years with a 35 pack-year smoking history presented with painless subacute paresthesia and weight fluctuation. A non-length-dependent small fibre neuropathy was confirmed by skin biopsy. Further testing revealed positive serum PCA-2 and CRMP-5 autoantibodies, which after positron emission tomography-CT led to histological confirmation of a small cell lung cancer. Initially, abnormal MRI and cerebrospinal fluid studies suggested central nervous system (CNS) involvement which was subclinical; however, 6â months later during antitumour therapy, the patient became symptomatic with choreoathetosis. After combined chemoradiation as well as immunosuppressive and symptomatic therapies, the clinical course stabilised, although residual neurological deficits remained at follow-up a year later. Coexistent PCA-2 and CRMP-5 autoantibodies may occur in the setting of small fibre peripheral neuropathy and choreoathetosis and predict cancer type. Two paraneoplastic syndromes can present successively over months; subclinical CNS involvement with evolving basal ganglia abnormalities can be a paraneoplastic manifestation. In the appropriate clinical setting, paraneoplastic testing should be considered in patients presenting with small fibre neuropathy.
Assuntos
Autoanticorpos/sangue , Coreia/complicações , Neoplasias Pulmonares/complicações , Proteínas do Tecido Nervoso/imunologia , Síndromes Paraneoplásicas/complicações , Células de Purkinje/imunologia , Carcinoma de Pequenas Células do Pulmão/complicações , Neuropatia de Pequenas Fibras/complicações , Diagnóstico Diferencial , Humanos , Hidrolases , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/terapia , Masculino , Proteínas Associadas aos Microtúbulos , Pessoa de Meia-Idade , Síndromes Paraneoplásicas/imunologia , Carcinoma de Pequenas Células do Pulmão/diagnóstico por imagem , Carcinoma de Pequenas Células do Pulmão/terapia , Resultado do TratamentoRESUMO
Neuron-specific enolase (NSE) is a glycolytic isoenzyme found in mature neurons and cells of neuronal origin. Injecting adeno-associated virus serotype 9 (AAV9) vectors carrying the NSE promoter into the cerebellar cortex is likely to cause the specific transduction of neuronal cells, such as Purkinje cells (PCs) and interneurons, but not Bergmann glia (BG). However, we found BG-predominant transduction without PC transduction along a traumatic needle tract for viral injection. The enhancement of neuroinflammation by the co-application of lipopolysaccharide (LPS) with AAV9 significantly expanded the BG-predominant area concurrently with the potentiated microglial activation. The BG-predominant transduction was gradually replaced by the PC-predominant transduction as the neuroinflammation dissipated. Experiments using glioma cell cultures revealed significant activation of the NSE promoter due to glucose deprivation, suggesting that intracellularly stored glycogen is metabolized through the glycolytic pathway for energy. Activation of the glycolytic enzyme promoter in BG concurrently with inactivation in PC may have pathophysiological significance for the production of lactate in activated BG and the utilization of lactate, which is provided by the BG-PC lactate shuttle, as a primary energy resource in injured PCs.
Assuntos
Inflamação/genética , Neuroglia/imunologia , Fosfopiruvato Hidratase/genética , Células de Purkinje/imunologia , Animais , Células Cultivadas , Córtex Cerebelar/imunologia , Córtex Cerebelar/metabolismo , Dependovirus/genética , Vetores Genéticos/administração & dosagem , Inflamação/induzido quimicamente , Ácido Láctico/metabolismo , Lipopolissacarídeos/efeitos adversos , Camundongos , Neuroglia/metabolismo , Regiões Promotoras Genéticas , Células de Purkinje/metabolismo , RatosRESUMO
IMPORTANCE: Classic Purkinje cell cytoplasmic antibody type 1 (PCA-1, or anti-Yo) paraneoplastic cerebellar ataxia has a poor prognosis, yet little has been published otherwise regarding treatment responses and outcomes among patients with autoimmune cerebellar ataxia. OBJECTIVES: To investigate treatment responses and outcomes in adults with autoimmune cerebellar ataxia. DESIGN, SETTING, AND PARTICIPANTS: A cohort study conducted at Mayo Clinic, Rochester, Minnesota, included 118 patients who had ataxia, were 18 years or older, were seropositive for at least 1 neural autoantibody, had received at least 1 immunotherapy or cancer therapy, and had neurologist-reported outcomes documented from January 1, 1989, through December 31, 2013. Data were collected from May 14, 2013, through August 9, 2014, and analyzed from August 9, 2014, through April 27, 2015. Responses to immunotherapy (corticosteroids, intravenous immunoglobulin, plasma exchange, and immunosuppressants) and ambulatory outcomes were compared between different subgroups. Subgroups were classified as paraneoplastic vs nonparaneoplastic disorders; neuronal nuclear and/or cytoplasmic (NNC) antibody positivity vs plasma membrane protein (PMP) antibody positivity; and glutamic acid decarboxylase 65-kDa isoform (GAD65) antibody positivity vs PMP antibody positivity. MAIN OUTCOMES AND MEASURES: Response to therapy and ambulatory ability, with univariate logistic regression and Kaplan-Meier analyses. RESULTS: Inclusion criteria were met by 118 patients. Median age at onset of neurologic symptoms was 58 (range, 27-83) years, and 87 patients (73.7%) were women. Median duration from symptom onset to last follow-up was 25 (range, 2-223) months. Sixty-three patients had paraneoplastic and 55 patients had nonparaneoplastic ataxic disorders. Eighty-one patients were seropositive for NNC antibodies (most commonly PCA-1 [anti-Yo], antineuronal nuclear antibody type 1 [anti-Hu], and GAD65 antibody); 22 patients, for neural PMP receptor or ion channel antibodies (most commonly targeting P/Q- or N-type voltage-gated calcium channels); and 15 patients, for antibodies from both categories. Neurologic improvements occurred in 54 patients (with a robust change in ambulatory ability in 22) attributable to immunotherapy; univariate regression analysis revealed that improvements were significantly more common among patients with nonparaneoplastic disorders (P = .03) and those with exclusively PMP antibodies (P = .02). Kaplan-Meier analyses revealed that progression to wheelchair dependence occurred significantly faster among patients with NNC antibody positivity only (P = .02), although those with GAD65 autoimmunity progressed to wheelchair dependence at a rate similar to those with PMP autoimmunity (P = .92). CONCLUSIONS AND RELEVANCE: Although autoimmune ataxia is usually severe, treatment responses can be gratifying, particularly in patients with nonparaneoplastic disorders and in those harboring autoantibodies directed against GAD65 or neural PMPs.
Assuntos
Autoanticorpos/sangue , Doenças Autoimunes do Sistema Nervoso , Ataxia Cerebelar , Imunoterapia/métodos , Avaliação de Resultados em Cuidados de Saúde , Degeneração Paraneoplásica Cerebelar , Adulto , Idoso , Idoso de 80 Anos ou mais , Autoanticorpos/líquido cefalorraquidiano , Doenças Autoimunes do Sistema Nervoso/imunologia , Doenças Autoimunes do Sistema Nervoso/fisiopatologia , Doenças Autoimunes do Sistema Nervoso/terapia , Ataxia Cerebelar/imunologia , Ataxia Cerebelar/fisiopatologia , Ataxia Cerebelar/terapia , Feminino , Glutamato Descarboxilase/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Degeneração Paraneoplásica Cerebelar/imunologia , Degeneração Paraneoplásica Cerebelar/fisiopatologia , Degeneração Paraneoplásica Cerebelar/terapia , Células de Purkinje/imunologiaRESUMO
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.
Assuntos
Autoanticorpos/imunologia , Neoplasias da Mama/imunologia , Morte Celular/imunologia , Neoplasias dos Genitais Femininos/imunologia , Proteínas do Tecido Nervoso/imunologia , Degeneração Paraneoplásica Cerebelar/imunologia , Células de Purkinje/imunologia , Animais , Neoplasias da Mama/patologia , Cerebelo/imunologia , Cerebelo/patologia , Feminino , Neoplasias dos Genitais Femininos/patologia , Humanos , Imunoglobulina G/imunologia , Degeneração Paraneoplásica Cerebelar/patologia , Células de Purkinje/patologia , Ratos , Ratos Sprague-DawleyRESUMO
Cerebellar ataxia commonly occurs in multiple sclerosis, particularly in chronic progressive disease. Previous reports have highlighted both white matter and grey matter pathological changes within the cerebellum; and demyelination and inflammatory cell infiltrates appear commonly. As Purkinje cell axons are the sole output of the cerebellar cortex, understanding pathologic processes within these cells is crucial to develop strategies to prevent their loss and thus reduce ataxia. We studied pathologic changes occurring within Purkinje cells of the cerebellum. Using immunohistochemic techniques, we found changes in neurofilament phosphorylation states within Purkinje cells, including loss of dephosphorylated neurofilament and increased phosphorylated and hyperphosphorylated neurofilament. We also found Purkinje axonal spheroids and Purkinje cell loss, both of which occurred predominantly within areas of leucocortical demyelination within the cerebellar cortex. These changes have important implications for the study of cerebellar involvement in multiple sclerosis and may help design therapies to reduce the burden of ataxia in the condition.
Assuntos
Cerebelo/patologia , Esclerose Múltipla/patologia , Células de Purkinje/patologia , Idoso , Idoso de 80 Anos ou mais , Axônios/imunologia , Axônios/metabolismo , Axônios/patologia , Morte Celular , Cerebelo/imunologia , Cerebelo/metabolismo , Estudos de Coortes , Feminino , Imunofluorescência , Humanos , Filamentos Intermediários/imunologia , Filamentos Intermediários/metabolismo , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/metabolismo , Fosforilação , Células de Purkinje/imunologia , Células de Purkinje/metabolismoAssuntos
Ataxia/etiologia , Doença Celíaca/complicações , Dieta Livre de Glúten , Leucoencefalopatias/etiologia , Transtornos Mentais/etiologia , Biópsia , Doença Celíaca/diagnóstico , Doença Celíaca/dietoterapia , Doença Celíaca/psicologia , Reações Cruzadas , Duodeno/patologia , Feminino , Deficiência de Ácido Fólico/complicações , Deficiência de Ácido Fólico/tratamento farmacológico , Lobo Frontal/fisiopatologia , Humanos , Deficiência de IgA/complicações , Ferro/uso terapêutico , Deficiências de Ferro , Leucoencefalopatias/diagnóstico por imagem , Leucoencefalopatias/fisiopatologia , Leucoencefalopatias/psicologia , Imageamento por Ressonância Magnética , Transtornos Mentais/fisiopatologia , Células de Purkinje/imunologia , Tremor/etiologia , Adulto JovemRESUMO
Paraneoplastic cerebellar degeneration (PCD) is characterized by loss of Purkinje cells (PCs) associated with progressive pancerebellar dysfunction in the presence of onconeural Yo antibodies. These antibodies recognize the cerebellar degeneration-related antigens CDR2 and CDR2L. Response to PCD therapy is disappointing due to limited understanding of the neuropathological mechanisms. Here, we report the pathological role of CDR antibodies on the calcium homeostasis in PCs. We developed an antibody-mediated PCD model based on co-incubation of cerebellar organotypic slice culture with human patient serum or rabbit CDR2 and CDR2L antibodies. The CDR antibody-induced pathology was investigated by high-resolution multiphoton imaging and biochemical analysis. Both human and rabbit CDR antibodies were rapidly internalized by PCs and led to reduced immunoreactivity of calbindin D28K (CB) and L7/Pcp-2 as well as reduced dendritic arborizations in the remaining PCs. Washout of the CDR antibodies partially recovered CB immunoreactivity, suggesting a transient structural change in CB calcium-binding site. We discovered that CDR2 and CB co-immunoprecipitate. Furthermore, the expression levels of voltage-gated calcium channel Cav2.1, protein kinase C gamma and calcium-dependent protease, calpain-2, were increased after CDR antibody internalization. Inhibition of these signaling pathways prevented or attenuated CDR antibody-induced CB and L7/Pcp-2 immunoreactivity loss, morphological changes and increased protein expression. These results signify that CDR antibody internalization causes dysregulation of cell calcium homeostasis. Hence, drugs that modulate these events may represent novel neuroprotective therapies that limit the damaging effects of CDR antibodies and prevent PC neurodegeneration.
Assuntos
Autoanticorpos/imunologia , Autoantígenos/imunologia , Cálcio/metabolismo , Proteínas do Tecido Nervoso/imunologia , Degeneração Paraneoplásica Cerebelar/imunologia , Células de Purkinje/imunologia , Idoso , Idoso de 80 Anos ou mais , Animais , Autoanticorpos/metabolismo , Autoantígenos/metabolismo , Calbindina 1/metabolismo , Canais de Cálcio Tipo N/metabolismo , Calpaína/metabolismo , Feminino , Homeostase/efeitos dos fármacos , Homeostase/fisiologia , Humanos , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas do Tecido Nervoso/metabolismo , Degeneração Paraneoplásica Cerebelar/patologia , Proteína Quinase C/metabolismo , Células de Purkinje/patologia , Coelhos , Ratos Wistar , Técnicas de Cultura de TecidosRESUMO
IMPORTANCE: Purkinje cell cytoplasmic antibody type 1 (PCA-1)-IgG (or anti-Yo) is characteristically detected in women with gynecological or breast adenocarcinoma. We describe 2 unique scenarios occurring in 1 patient: PCA-1 paraneoplastic autoimmunity in a child, and a paraneoplastic neurological disorder in the context of Down syndrome. OBSERVATIONS: A child with Down syndrome and a history of adrenocortical carcinoma resected at age 1 year presented at age 7 years with cerebellar ataxia of subacute onset. Paraneoplastic serological and cerebrospinal fluid evaluations revealed PCA-1. Serological and biochemical studies also supported a diagnosis of subclinical autoimmune hypothyroidism. Extensive serum, urine, and radiological testing did not reveal a new or recurrent neoplasm. Neurological improvements after standard immunotherapy were lacking. CONCLUSIONS AND RELEVANCE: Solid organ neoplasms are uncommon among patients with Down syndrome, but organ-specific autoimmune diseases are common. In our patient, Down syndrome-related impaired T regulatory lymphocyte function (previously reported) may have resulted in both enhanced immunity against an undetected solid neoplasm and paraneoplastic neurological (PCA-1) autoimmunity.