Alterações de líquor e imunofenotipagem linfocítica em modelos experimentais de Neurocisticercose Extraparenquimatosa
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Data
2021-08-26
Autores
Alves Junior, Aderaldo Costa
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Universidade Estadual Paulista (Unesp)
Resumo
A Neurocisticercose (NCC) é uma doença de grande prevalência mundial, levando a um alto impacto social e na saúde pública. A forma extraparenquimatosa da doença ocorre quando o cisticerco está presente no espaço subaracnóideo dos pacientes, provocando uma reação inflamatória no líquor (LCR) que cursa com grave sintomatologia e altas taxas de morbimortalidade. Modelos experimentais de NCC têm grande importância por permitir o melhor entendimento da doença e assim otimizar a terapêutica. Objetivamos neste estudo analisar as alterações no líquor e sistema nervoso central (SNC) de animais infectados com Taenia crassiceps em seus espaços subaracnoideos, bem como caracterizar as linhagens linfocitárias envolvidas na resposta inflamatória cerebral. Na primeira parte do estudo, 39 coelhos foram submetidos à inoculação de cistos (grupo 1, ninicial=20, nfinal=13) ou antígenos (grupo 2, ninicial=19, nfinal=12) de T. crassiceps por meio de punção das cisternas magnas, sendo observados por 6 meses com coletas periódicas de LCR para análise laboratorial, imagens de ressonância magnética (RM) e remoção encefálica para análise histopatológica. Os coelhos apresentaram aumento de celularidade e proteinorraquia no líquor com pico em duas semanas e recuperação em até três meses, sendo mais significativos no grupo 1 (cistos), além de modificação no padrão celular liquórico para um predomínio linfocitário persistente. Houve também aumento nos volumes ventriculares (hidrocefalia presente em 64,3% dos animais do grupo 1 e 55,6% do grupo 2) e na cisterna quadrigeminal (alargamento cisternal presente em 92,9% dos animais do grupo 1 e 66,7% do grupo 2) nas imagens de RM, enquanto que a análise histopatológica evidenciou padrão de meningite linfocitária discreta na maioria das lâminas de ambos os grupos e uma densidade de neurônios inferior no grupo inoculado com cistos (grupo 1) em relação ao grupo 2 (antígenos). O modelo experimental mostrou-se viável e prático em coelhos, reproduzindo as alterações observadas na NCC humana. Já na segunda parte do projeto, a imunofenotipagem do tecido cerebral para determinação do padrão linfocitário foi realizada em 19 ratos expostos a cistos de T. crassiceps em seus espaços subaracnoideos por: 1 mês (grupo 1R, n=4), 4 meses (grupo 2R, n=11) ou 6 meses (grupo 3R, n=4). Foi encontrada expressão para os marcadores linfocitários CD19, CD56, CD8 e CD4, sendo o último em uma apresentação bimodal com picos no 1º e 6º meses após as inoculações, e o CD8 com maior expressão após 6 meses de observação.
Neurocysticercosis (NCC) is a highly prevalent disease worldwide, leading to an important social and public health impact. The extraparenchymal form occurs when the cysticercus is located in the subarachnoid space, causing an inflammatory reaction to the cerebrospinal fluid (CSF) that evolves with severe symptoms and high rates of morbidity and mortality. Experimental models of NCC are crucial for allowing a better understanding of the disease and thus optimizing therapy. The aim of this study was to analyze changes in the CSF and central nervous system (CNS) of animals infected with Taenia crassiceps in their subarachnoid spaces, as well as to characterize the lymphocyte strains involved in the CNS inflammatory response. In the first part of this study, 39 rabbits were inoculated with cysts (group 1, ni=20, n=13), or antigens (group 2, ni=19, n=12) of T. crassiceps through punctures to the cisterna magna, followed by observation for 6 months with periodic CSF samples collection for laboratory analysis, magnetic resonance imaging (MRI) and finally brain removal for histopathological analysis. These animals showed increased cellularity and protein concentration in CSF with a peak in two weeks and normalization in up to three months, more significant in the group 1 (cysts), in addition to changes in CSF cell pattern for a persistent lymphocyte predominance. There was also an increase in ventricular (60.9% of animals) and quadrigeminal cistern (82.6%) volumes in both groups on MRI, while histopathology showed a mild lymphocytic meningitis on most of the cuts in both groups and an inferior density of neurons on group 1 compared to group 2. This experimental model proved to be viable in rabbits, reproducing the changes observed in human NCC. In the second part of this project, immunophenotyping of brain tissue to determine the lymphocyte pattern was performed in 19 rats exposed to cysts of T. crassiceps in their subarachnoid spaces for: 1 month (group 1R, n=4), 4 months (group 2R, n=11) or 6 months (group 3R, n=4). We found expression to the lymphocytic markers CD19, CD56, CD8 and CD4, the latter with a bimodal fashion with peaks in the 8 first and sixth months after the inoculations, and CD8 presenting higher expression after six months of observational period.
Neurocysticercosis (NCC) is a highly prevalent disease worldwide, leading to an important social and public health impact. The extraparenchymal form occurs when the cysticercus is located in the subarachnoid space, causing an inflammatory reaction to the cerebrospinal fluid (CSF) that evolves with severe symptoms and high rates of morbidity and mortality. Experimental models of NCC are crucial for allowing a better understanding of the disease and thus optimizing therapy. The aim of this study was to analyze changes in the CSF and central nervous system (CNS) of animals infected with Taenia crassiceps in their subarachnoid spaces, as well as to characterize the lymphocyte strains involved in the CNS inflammatory response. In the first part of this study, 39 rabbits were inoculated with cysts (group 1, ni=20, n=13), or antigens (group 2, ni=19, n=12) of T. crassiceps through punctures to the cisterna magna, followed by observation for 6 months with periodic CSF samples collection for laboratory analysis, magnetic resonance imaging (MRI) and finally brain removal for histopathological analysis. These animals showed increased cellularity and protein concentration in CSF with a peak in two weeks and normalization in up to three months, more significant in the group 1 (cysts), in addition to changes in CSF cell pattern for a persistent lymphocyte predominance. There was also an increase in ventricular (60.9% of animals) and quadrigeminal cistern (82.6%) volumes in both groups on MRI, while histopathology showed a mild lymphocytic meningitis on most of the cuts in both groups and an inferior density of neurons on group 1 compared to group 2. This experimental model proved to be viable in rabbits, reproducing the changes observed in human NCC. In the second part of this project, immunophenotyping of brain tissue to determine the lymphocyte pattern was performed in 19 rats exposed to cysts of T. crassiceps in their subarachnoid spaces for: 1 month (group 1R, n=4), 4 months (group 2R, n=11) or 6 months (group 3R, n=4). We found expression to the lymphocytic markers CD19, CD56, CD8 and CD4, the latter with a bimodal fashion with peaks in the 8 first and sixth months after the inoculations, and CD8 presenting higher expression after six months of observational period.
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Palavras-chave
Taenia, Neurocisticercose, Imunofenotipagem, Líquido cefalorraquidiano, Cisticercose