RESUMO
Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and little is known about the occurrence and pathogenesis of this parasite in the CNS. The aims of this study were to evaluate the occurrence, viability and load of L. infantum in the CNS, and to identify the neurological histological alterations associated with this protozoan and its co-infections in naturally infected dogs. Forty-eight Leishmania-seropositive dogs from which L. infantum was isolated after necropsy were examined. Cerebrospinal fluid (CSF) samples were analyzed by parasitological culture, quantitative real-time PCR (qPCR) and the rapid immunochromatographic Dual Path Platform test. Brain, spinal cord and spleen samples were submitted to parasitological culture, qPCR, and histological techniques. Additionally, anti-Toxoplasma gondii and anti-Ehrlichia canis antibodies in serum and distemper virus antigens in CSF were investigated. None of the dogs showed neurological signs. All dogs tested positive for L. infantum in the CNS. Viable forms of L. infantum were isolated from CSF, brain and spinal cord in 25% of the dogs. Anti-L. infantum antibodies were detected in CSF in 61% of 36 dogs. Inflammatory histological alterations were observed in the CNS of 31% of the animals; of these, 66% were seropositive for E. canis and/or T. gondii. Amastigote forms were associated with granulomatous non-suppurative encephalomyelitis in a dog without evidence of co-infections. The highest frequency of L. infantum DNA was observed in the brain (98%), followed by the spinal cord (96%), spleen (95%), and CSF (50%). The highest L. infantum load in CNS was found in the spinal cord. These results demonstrate that L. infantum can cross the blood-brain barrier, spread through CSF, and cause active infection in the entire CNS of dogs. Additionally, L. infantum can cause inflammation in the CNS that can lead to neurological signs with progression of the disease.
Assuntos
Doenças do Sistema Nervoso Central/veterinária , Doenças do Cão/parasitologia , Leishmania infantum/fisiologia , Leishmaniose Visceral/veterinária , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Sistema Nervoso Central/parasitologia , Sistema Nervoso Central/patologia , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Doenças do Sistema Nervoso Central/parasitologia , Coinfecção/microbiologia , Coinfecção/parasitologia , Coinfecção/veterinária , DNA de Protozoário/genética , Doenças do Cão/microbiologia , Cães , Ehrlichia canis/imunologia , Ehrlichia canis/fisiologia , Ehrlichiose/microbiologia , Ehrlichiose/veterinária , Interações Hospedeiro-Parasita , Interações Hospedeiro-Patógeno , Hibridização In Situ , Leishmania infantum/genética , Leishmania infantum/imunologia , Leishmaniose Visceral/líquido cefalorraquidiano , Leishmaniose Visceral/parasitologia , Carga Parasitária , Reação em Cadeia da Polimerase em Tempo Real , Toxoplasma/imunologia , Toxoplasma/fisiologia , Toxoplasmose/parasitologiaRESUMO
To investigate the cytokine profile in serum and cerebrospinal fluid (CSF) from patients with systemic lupus erythematosus (SLE) and central nervous system infection, we measured interferon-g (IFN-g), interleukin-1b (IL-1b), IL-4, IL-6, IL-8, IL-10, and IL-17 levels in serum and CSF from 50 SLE patients and 38 matched controls. In patients with active compared to quiescent disease, serum levels were higher for IL-1b (P = 0.042) and IL-17 (P = 0.041) but we found no significant correlation between IL-1b and IL-17 and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) (r = 0.055, r = 0.219, respectively). IL-10 level in active patients was lower compared to that in quiescent controls (P = 0.032). When comparing specific disease manifestations, IL-1b levels in patients with fever (P = 0.035) and IL-6 (P = 0.048) and IL-8 (P = 0.048) levels in those showing nervous system involvement were higher than in controls. Based on MRI results, we found that only increased cerebral ischemia was associated with increased IFN-g levels (P = 0.009). In neuropsychiatric lupus erythematous patients, CSF levels of IL-6 (P = 0.002), IL-8 (P = 0.009), and IL-17 (P = 0.034) were significantly higher when compared with control patients. IL-10:IL-1b ratio in patients with moderate and quiescent disease was higher than in patients with disease activity (P = 0.000). Pro-inflammatory adaptive cytokines were elevated during disease flare, while regulatory mediators were elevated during periods of stable disease. Alterations in the balance between inflammatory and regulatory mediators may be targets for novel immunotherapeutic agents for managing autoimmune diseases.
Assuntos
Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Interleucina-17/líquido cefalorraquidiano , Interleucina-6/líquido cefalorraquidiano , Interleucina-8/líquido cefalorraquidiano , Lúpus Eritematoso Sistêmico/líquido cefalorraquidiano , Idoso , Doenças do Sistema Nervoso Central/sangue , Doenças do Sistema Nervoso Central/diagnóstico por imagem , Doenças do Sistema Nervoso Central/patologia , Feminino , Humanos , Interferon gama/sangue , Interferon gama/líquido cefalorraquidiano , Interleucina-10/sangue , Interleucina-10/líquido cefalorraquidiano , Interleucina-17/sangue , Interleucina-1beta/sangue , Interleucina-1beta/líquido cefalorraquidiano , Interleucina-4/sangue , Interleucina-4/líquido cefalorraquidiano , Interleucina-6/sangue , Interleucina-8/sangue , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/diagnóstico por imagem , Lúpus Eritematoso Sistêmico/patologia , Masculino , Pessoa de Meia-IdadeRESUMO
The cell count and differential of cerebrospinal fluid (CSF) cytologic examination classify CSF as inflammatory or not. The cytospin cell yield is related to cell count, but to our knowledge a relationship has not been characterized and cytospin precision is undocumented in any species. The objective of our study was to calculate intra-assay precision of cellular yield and differential on cytocentrifuged canine CSF, determine the factors that may affect precision, and predict the number of cytospins necessary to confirm mild neutrophilic pleocytosis. Ten concurrent replicate cytospins were created from nonhemorrhagic CSF, obtained from 60 dogs in other terminal studies, with either a manual or calibrated pipetting technique. Up to 500 cells per cytospin were counted and classified on each slide. Coefficient of variation (CV), multiple regression, and probabilities were calculated for relationships between cell yield and independent factors including technique, total nucleated cell count, cell differential, and total protein. Manual and calibrated pipetting had similar CVs (average 31%) for total cell yield, but the calibrated technique had fewer foamy macrophages. CV for neutrophil percentage among low cellularity samples with any neutrophils was 146%. Probability based on linear regression showed that 1 cytospin is sufficient to identify samples with >3% neutrophils. Occasional neutrophils, eosinophils, mitotic figures, phagocytic cells, and ependymal cells were seen in many low cellularity canine CSF samples. Canine CSF cytospin cell yield and differential evaluations are imprecise. Calibrated rather than manual pipetting is recommended.
Assuntos
Doenças do Sistema Nervoso Central/veterinária , Líquido Cefalorraquidiano/citologia , Doenças do Cão/líquido cefalorraquidiano , Animais , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Cães , Sensibilidade e EspecificidadeRESUMO
Diagnostic lumbar puncture is essential to the diagnosis of central nervous system infections, subarachnoid haemorrhage and others neurological diseases. Myeloradicular involvement or life-threatening adverse events due to the procedure are rare, but less severe complications are more frequent. Post-lumbar puncture headache is the most common complication, by spinal fluid leakage due to delayed closure of a dural defect. Therefore, the development of fine needles, with differentiated atraumatic bevel, has contributed to minimize that problem. These generically called atraumatic needles cause less deformation of the dura mater then the Quincke(®) ones. So, why don't we use these atraumatic needles?
Assuntos
Doenças do Sistema Nervoso Central/diagnóstico , Agulhas , Punção Espinal/instrumentação , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Desenho de Equipamento , Humanos , Cefaleia Pós-Punção Dural , Punção Espinal/efeitos adversosRESUMO
Diagnostic lumbar puncture is essential to the diagnosis of central nervous system infections, subarachnoid haemorrhage and others neurological diseases. Myeloradicular involvement or life-threatening adverse events due to the procedure are rare, but less severe complications are more frequent. Post-lumbar puncture headache is the most common complication, by spinal fluid leakage due to delayed closure of a dural defect. Therefore, the development of fine needles, with differentiated atraumatic bevel, has contributed to minimize that problem. These generically called atraumatic needles cause less deformation of the dura mater then the Quincke® ones. So, why don't we use these atraumatic needles?.
A punção lombar é essencial para o diagnóstico de infecções do sistema nervoso central, hemorragia subaracnoídea e outras doenças neurológicas. O comprometimento mielorradicular ou efeitos adversos que envolvam risco de vida como consequência do procedimento são raros, mas complicações leves são mais frequentes. Cefaléia pós-punção lombar é a complicação mais comum, secundária a escoamento do líquido cefalorraquiano por fechamento tardio de uma lesão dural. Consequentemente, o desenvolvimento de agulhas finas, com bisel atraumático, tem contribuído para minimizar o problema. Estas agulhas, chamadas genericamente de atraumáticas, causam menos deformação da dura-mater que as agulhas tipo Quincke®. Então, porque não usar estas agulhas atraumáticas?.
Assuntos
Humanos , Doenças do Sistema Nervoso Central/diagnóstico , Agulhas , Punção Espinal/instrumentação , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Desenho de Equipamento , Cefaleia Pós-Punção Dural , Punção Espinal/efeitos adversosRESUMO
The IgG index measures the intrathecal immunoglobulin production and it is a useful tool for diagnosis of inflammatory diseases involving the central nervous system. This index is based on the precise quantification of albumin and IgG in canine cerebrospinal fluid and serum. Here, we report the development of an indirect competitive ELISAs for the detection of both antigens. Thirty-two dogs were included in this study, divided into three experimental groups. Group A was composed of 22 healthy animals, as determined by standard clinical examination. In group B, six animals, presented neurological pathologies associated with endogenous IgG production and, in group C four animals presented neurological diseases or symptoms not associated with intrathecal IgG production. Cerebrospinal fluid and serum samples were obtained from these animals. As expected, by using the indirect ELISAs proposed here, the IgG indexes obtained in healthy animals (A) were 0.371+/-0.252 (SD). In B and C, the values (3.002+/-1.897; 0.36+/-0.306, respectively), were in agreement with the pathologic conditions of the individuals in each group. Thus, the immunometric competition ELISA methods proposed here allow the discrimination of abnormal intrathecal IgG production, in a variety of inflammatory pathologic conditions of the central nervous system.
Assuntos
Doenças do Sistema Nervoso Central/veterinária , Doenças do Cão/imunologia , Imunoglobulina G/análise , Inflamação/veterinária , Animais , Doenças do Sistema Nervoso Central/sangue , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Doenças do Sistema Nervoso Central/imunologia , Doenças do Cão/sangue , Doenças do Cão/líquido cefalorraquidiano , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/veterinária , Técnicas Imunoenzimáticas/veterinária , Imunoglobulina G/sangue , Imunoglobulina G/líquido cefalorraquidiano , Inflamação/sangue , Inflamação/líquido cefalorraquidiano , Inflamação/imunologia , Coelhos/imunologia , Valores de Referência , Sensibilidade e Especificidade , Albumina Sérica/metabolismoRESUMO
BACKGROUND: In the central nervous system, HIV replication can occur relatively independent of systemic infection, and intrathecal replication of HIV-1 has been observed in patients with HIV-related and opportunistic neurological diseases. The clinical usefulness of HIV-1 RNA detection in the cerebrospinal fluid (CSF) of patients with opportunistic neurological diseases, or the effect of opportunistic diseases on CSF HIV levels in patients under HAART has not been well defined. We quantified CSF and plasma viral load in HIV-infected patients with and without different active opportunistic neurological diseases, determined the characteristics that led to a higher detection rate of HIV RNA in CSF, and compared these two compartments. METHODS: A prospective study was conducted on 90 HIV-infected patients submitted to lumbar puncture as part of a work-up for suspected neurological disease. Seventy-one patients had active neurological diseases while the remaining 19 did not. RESULTS: HIV-1 RNA was quantified in 90 CSF and 70 plasma samples. The HIV-1 RNA detection rate in CSF was higher in patients with neurological diseases, in those with a CD4 count lower than 200 cells/mm3, and in those not receiving antiretroviral therapy, as well as in patients with detectable plasma HIV-1 RNA. Median viral load was lower in CSF than in plasma in the total population, in patients without neurological diseases, and in patients with toxoplasmic encephalitis, while no significant difference between the two compartments was observed for patients with cryptococcal meningitis and HIV-associated dementia. CSF viral load was lower in patients with cryptococcal meningitis and neurotoxoplasmosis under HAART than in those not receiving HAART. CONCLUSION: Detection of HIV-1 RNA in CSF was more frequent in patients with neurological disease, a CD4 count lower than 200 cells/mm3 and detectable plasma HIV-1. Median HIV-1 RNA levels were generally lower in CSF than in plasma but some patients showed higher CSF levels, and no difference between these two compartments was observed in patients with cryptococcal meningitis and HIV-associated dementia, suggesting the presence of intrathecal viral replication in these patients. HAART played a role in the control of CSF HIV levels even in patients with cryptococcal meningitis and neurotoxoplasmosis in whom viral replication is potentially higher.
Assuntos
Complexo AIDS Demência/virologia , Infecções Oportunistas Relacionadas com a AIDS/virologia , Antirretrovirais/uso terapêutico , Terapia Antirretroviral de Alta Atividade , Doenças do Sistema Nervoso Central/virologia , HIV-1/fisiologia , RNA Viral/sangue , RNA Viral/líquido cefalorraquidiano , Complexo AIDS Demência/sangue , Complexo AIDS Demência/líquido cefalorraquidiano , Infecções Oportunistas Relacionadas com a AIDS/sangue , Infecções Oportunistas Relacionadas com a AIDS/líquido cefalorraquidiano , Adulto , Contagem de Linfócito CD4 , Doenças do Sistema Nervoso Central/sangue , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Feminino , Humanos , Masculino , Meningite Criptocócica/sangue , Meningite Criptocócica/líquido cefalorraquidiano , Meningite Criptocócica/virologia , Pessoa de Meia-Idade , Estudos Prospectivos , RNA Viral/efeitos dos fármacos , Toxoplasmose Cerebral/sangue , Toxoplasmose Cerebral/líquido cefalorraquidiano , Toxoplasmose Cerebral/virologia , Carga Viral , Replicação ViralAssuntos
Humanos , Masculino , Feminino , Recém-Nascido , Lactente , Pré-Escolar , Criança , Glucose/líquido cefalorraquidiano , Líquido Cefalorraquidiano/química , Meningites Bacterianas/líquido cefalorraquidiano , Cardiopatias Congênitas/líquido cefalorraquidiano , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Febre/líquido cefalorraquidiano , Leucemia/líquido cefalorraquidiano , Líquido Cefalorraquidiano/microbiologia , Valores de ReferênciaRESUMO
We studied 53 samples of cerebrospinal fluid (CSF) by cytologic examination and immunophenotyping by flow cytometry. The samples were taken from 43 patients; 25 had a previous diagnosis of malignant lymphoma/leukemia and the remaining 18 a variety of other diseases involving the central nervous system (CNS). Lymphoma/leukemia was detected in 21 samples: 12 by morphologic examination and immunophenotyping and nine by immunophenotyping alone. There were two cases with a suspicious morphologic examination and negative immunophenotyping in which the final diagnosis were cryptococcal and viral meningitis. In the group of 18 patients, one was diagnosed as a primary malignant lymphoma of the CNS and was positive with cytology and immunophenotyping. The other 17 were negative with both methods and follow-up showed no evidence of lymphoma/leukemia. This study shows that morphologic examination combined with flow cytometry enhances the detection rate by 75% over morphologic examination alone in CSF samples.
Assuntos
Líquido Cefalorraquidiano/citologia , Leucemia/líquido cefalorraquidiano , Linfoma/líquido cefalorraquidiano , Adolescente , Adulto , Idoso , Antígenos CD/metabolismo , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Doenças do Sistema Nervoso Central/patologia , Criança , Citodiagnóstico/métodos , Feminino , Citometria de Fluxo/métodos , Humanos , Imunofenotipagem , Leucemia/patologia , Linfoma/patologia , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
We investigated the levels of prolactin (PRL) and interleukin-6 (IL-6) in the cerebrospinal fluid (CSF) and serum of systemic lupus erythematosus patients with central nervous system involvement (CNS-SLE), and examined whether PRL and IL-6 have a relationship. Serum and CSF PRL and IL-6 were measured in the following groups of patients and controls: group I: seven patients with CNS-SLE; group II: three SLE patients without CNS involvement (non CNS-SLE); group III: 10 patients with neurocysticercosis; and group IV: six healthy women. The patients were clinically assessed. CSF PRL and IL-6 were elevated in group I (CNS-SLE) in comparison with all other groups (p<0.001). In addition, four of seven patients had higher levels of IL-6 and PRL in CSF than in serum. A positive correlation between PRL and IL-6 in CSF of SLE was observed (r=0.88, p<0.001). The mean serum PRL concentrations were not significantly different in all groups, but high levels of IL-6 were found in the serum of group I in comparison with groups II and IV (p<0.001). The serum levels of group III were not different from those of group I. These results demonstrate the presence of intrathecal synthesis and elevations of CSF PRL and IL-6 in active CNS-SLE involvement and indicate that measurements of CSF PRL and IL-6 may be useful in the evaluation of neuropsychiatric lupus erythematosus.