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Estudo da heterogeneidade de monócitos de cães infectados ou não por Leishmania infantum

Authors: Vitor Silva Barbosa;

Estudo da heterogeneidade de monócitos de cães infectados ou não por Leishmania infantum

Abstract

Monócitos do sangue periférico compõem o maior grupo de células progenitoras circulantes. Suas funções vêm sendo reveladas com o estudo de suas subpopulações, especialmente em humanos e camundongos, em processos patológicos provenientes de doenças crônicas não infecciosas ou infecciosas transmissíveis. Dentre essas doenças estão as leishmanioses, das quais a mais grave é a forma visceral, caudadas por protozoários do gênero Leishmania. Esses são parasitos intracelulares obrigatórios em células do sistema fagocítico mononuclear do hospedeiro mamífero, como monócitos, macrófagos e células dendríticas. O cão (Canis familiares) representa o principal reservatório desse parasito, tendo papel central na transmissão a humanos. No Brasil, o parasito Leishmania (Leishmania) chagasi (sin. L. infantum) é o principal causador da leishmaniose visceral canina (LVA). Esse estudo objetivou caracterizar subpopulações de monócitos do sangue periférico de cães naturalmente infectados ou não infectados por L. infantum e comparar seus aspectos fenotípicos e funcionais. Amostras de sangue periférico foram colhidas de cães de diferentes grupos: não infectados (n=5) e infectados (assintomáticos (n=5) e sintomáticos (n=5). As células foram marcadas por anticorpos contra as moléculas: CCR2, CD11a, CD11b, CD11c, CD14, CD16, CD44, CD45RO, CD56, CD62L, CD80, CX3CR1 e MHC-II e analisadas por citometria de fluxo. Células CD14+ constituem cerca de 4% da população de leucócitos totais e quando avaliamos essas células observamos duas populações distintas de acordo com a expressão de CD14: uma com alta expressão de CD14 (CD14hi), constituindo cerca de 90% dos monócitos, apresentando núcleo basofílico em forma de ferradura, com maior capacidade fagocítica; e outra com baixa expressão de CD14 (CD14lo), constituindo cerca de 10% dos monócitos, com núcleo de aspecto menos basofílico em forma de rim. Na análise das demais moléculas de superfície observou-se diferenças na expressão entre as populações. Em cães com LV não há diferença de frequência entre as populações, todavia há aumento da frequência de células CD14+ totais em cães infectados. Notamos também acentuada redução da expressão de moléculas relacionadas à ativação dos monócitos como CCR2, CD11c, CD45RO, CD62L e MHC-II, principalmente no grupo de cães sintomáticos, indicando possível inibição de sua ação no controle da doença.

Peripheral blood monocytes are the largest group of circulating progenitor cells. Its functions have been revealed through the study of their subpopulations, especially in humans and mice in pathological processes from infectious or noninfectious chronic diseases. Among the studied infectious diseases there is the leishmaniasis, which the most severe is the visceral leishmaniasis. Protozoa of the genus Leishmania are obligate intracellular parasites in cells of the mononuclear phagocyte system of the mammalian host, as monocytes, macrophages and dendritic cells. The dog (Canis familiar) is the main reservoir of this parasite, having a central role in the transmission to humans. In Brazil, the parasite Leishmania (Leishmania) chagasi (syn. L. infantum) is the main cause of the canine visceral leishmaniasis. This study aimed to characterize subpopulations of peripheral blood monocytes from uninfected or L. infantum naturally infected dogs and compare their phonotypical and functional aspects. Blood samples were collected from dogs from different groups: uninfected (n=5) and infected (asymptomatic (n=5) and symptomatic (n=5) with L. infantum. Cells were labeled by antibodies against the molecules: CCR2, CD11a, CD11b, CD11c, CD14, CD16, CD44, CD45RO, CD56, CD62L, CD80, CX3CR1 e MHC-II and the samples were obtained by flow cytometry. CD14+ cells represented about 4% of the total leucocytes and when analyzing those cells, it was observed two distinct populations according to CD14 expression: a population with high expression of CD14 (CD14hi), constituting about 90% of the monocytes showing basophilic and horseshoe-shaped nucleus with greater phagocytic capacity, and another with low expression of CD14 (CD14lo), constituting about 10% of monocytes, showing less basophilic and kidney-shaped nucleus. The analysis of the other molecules showed different levels of expression between the two populations. In dogs with visceral leishmaniasis no difference in frequency was observed between populations, however there is an increase of CD14+ cells in infected dogs. We also noticed marked reduction of the expression of molecules related to the activation of monocytes (CCR2, CD11c, CD45RO, CD62L and MHC-II), especially in the group of symptomatic dogs, indicating a possible inhibition on the capacity of these cells to control the disease.

Keywords

Leishmania, Heterogeneidade genética, Cão, L infantum, Marcadores genéticos, Monócitos, Sangue, Cães, Leishmaniose, Leishmania infantum, Doenças do cão/prevenção & controle, Patologia

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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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