Avaliação dos polimorfismos HPA em plaquetas expostas in vitro ao ZIKV
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Data
2021-05-27
Autores
Belo, Lucas Guerchi
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Universidade Estadual Paulista (Unesp)
Resumo
O ZIKA vírus é um arbovírus da família Flaviviridae e gênero flavivirus, possuindo grandes semelhanças genéticas e estruturais a outros arbovírus relevantes na área da saúde. A infecção por este patógeno está associada a complicações neurológicas graves, como microcefalia em neonatos e a síndrome de Guillain-Barre. Sua principal forma de transmissão é vetorial, sendo o mosquito fêmea do gênero Aedes o principal vetor. Contudo, outras vias de transmissão também foram documentadas, como a transmissão por transfusão de hemocomponentes, entre eles, a transfusão de concentrado de plaquetas. As plaquetas são fragmentos de megacariócitos que expressam diversas moléculas de superfície, as quais já foram associadas como receptores virais. Os Antígenos Plaquetários Humanos (HPA), por exemplo, são glicoproteínas de membrana presentes na superfície das plaquetas e estão associados a interações virais. Tais antígenos são suscetíveis a ocorrência de polimorfismos, e a presença desses polimorfismos estão associadas a infecções de flavivirus, tais como a dengue e Hepatite C. No entanto, ainda não existem estudos avaliando essa relação com ZIKV. Desse modo, o objetivo deste trabalho foi avaliar a possível associação entre os polimorfismos dos sistemas HPA -1, -3 e -5 com a capacidade do ZIKA vírus de interagir com plaquetas. As plaquetas foram obtidas de doadores, elas foram incubadas in vitro com ZIKV e em seguida lavadas. A presença do ZIKV nas plaquetas foi detectada por RT-qPCR e a genotipagem do HPA-1 e HPA-3 foram feitas por PCR-SSP, e do HPA-5 por PCR-RFLP. As amostras apresentaram carga viral detectadas nas plaquetas. Na análise estatística foi identificada significância entre a genotipagem do sistema HPA-1 e a capacidade do ZIKV de interagir com as plaquetas. Nossos dados sugerem um mecanismo de interação entre ZIKV e plaquetas, e uma associação entre o perfil HPA-1 e a afinidade do ZIKV interagir com as plaquetas.
ZIKA virus is an arbovirus of the family Flaviviridae and genus flavivirus. It has great genetic and structural similarities to other relevant arboviruse. This pathogen infection is associated with serious neurological complications, including microcephaly in neonates and Guillain-Barre syndrome. Its main form of transmission is through the female mosquito of the genus Aedes as a vector. However, other transmission routes have also been documented, such as transmission by transfusion of blood components, including the transfusion of platelet concentrate. Platelets are fragments of megakaryocytes that express several surface molecules, which have already been associated as viral receptors. Human Platelet Antigens (HPA), for example, are membrane glycoproteins present on the surface of platelets and are associated with viral interactions. Such antigens are susceptible to the occurrence of polymorphisms, and its presence is associated with flavivirus diseases, such as dengue and Hepatitis C. However, there are yet no studies evaluating this relationship with ZIKV. Therefore, the aim of this research has been to evaluate a possible association between the polymorphisms of the HPA -1, -3 and -5 systems and the ability of ZIKA virus to interact with platelets. Platelets were obtained from donors, incubated in vitro with ZIKV and then washed. The presence of ZIKV in the platelets was detected using RT-qPCR and genotyping of HPA-1 and HPA-3 has been performed through PCR-SSP, and HPA-5 by PCR-RFLP. Viral load was detected in platelets. Statistical analysis identified significance between the genotyping of the HPA-1 system and the ability of ZIKV to interact with platelets. Our updated data provided an interaction mechanism between ZIKV and platelets, as well as an association between the HPA-1 profile and the affinity of ZIKV interacting with platelets.
ZIKA virus is an arbovirus of the family Flaviviridae and genus flavivirus. It has great genetic and structural similarities to other relevant arboviruse. This pathogen infection is associated with serious neurological complications, including microcephaly in neonates and Guillain-Barre syndrome. Its main form of transmission is through the female mosquito of the genus Aedes as a vector. However, other transmission routes have also been documented, such as transmission by transfusion of blood components, including the transfusion of platelet concentrate. Platelets are fragments of megakaryocytes that express several surface molecules, which have already been associated as viral receptors. Human Platelet Antigens (HPA), for example, are membrane glycoproteins present on the surface of platelets and are associated with viral interactions. Such antigens are susceptible to the occurrence of polymorphisms, and its presence is associated with flavivirus diseases, such as dengue and Hepatitis C. However, there are yet no studies evaluating this relationship with ZIKV. Therefore, the aim of this research has been to evaluate a possible association between the polymorphisms of the HPA -1, -3 and -5 systems and the ability of ZIKA virus to interact with platelets. Platelets were obtained from donors, incubated in vitro with ZIKV and then washed. The presence of ZIKV in the platelets was detected using RT-qPCR and genotyping of HPA-1 and HPA-3 has been performed through PCR-SSP, and HPA-5 by PCR-RFLP. Viral load was detected in platelets. Statistical analysis identified significance between the genotyping of the HPA-1 system and the ability of ZIKV to interact with platelets. Our updated data provided an interaction mechanism between ZIKV and platelets, as well as an association between the HPA-1 profile and the affinity of ZIKV interacting with platelets.
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Palavras-chave
Zika vírus, Plaquetas, Human platelet antigen (HPA), Polimorfismo, Platelets, Human platelet antigen (HPA), Polymorphism