de Lima Cavalcanti Thaíse Yasmine Vasconcelos, Azevedo Elisa de Almeida Neves, Lima Morganna Costa, Saraiva Karina Lidiane Alcântara, Franca Rafael Freitas Oliveira
Department of Virology and Experimental Therapy, Instituto Aggeu Magalhães, Fundação Oswaldo Cruz/Fiocruz, Recife, Brazil.
Core Technology Platforms, Instituto Aggeu Magalhães, Fundação Oswaldo Cruz/Fiocruz, Recife, Brazil.
Front Microbiol. 2023 May 11;14:1152480. doi: 10.3389/fmicb.2023.1152480. eCollection 2023.
Chikungunya virus (CHIKV) is an arthropod-borne virus recently associated with large outbreaks in many parts of the world. Infection is typically manifested as a febrile and self-limited illness, characterized by joint pain and myalgia, albeit severe neurological manifestations are also reported. Although CHIKV is not recognized as a truly neurotropic virus, neurons, astrocytes, and oligodendrocytes are susceptible to infection . Here we employed a model of 3D cell culture to obtain neurospheres from ATRA/BNDF differentiated human neuroblastoma cells. We demonstrate that CHIKV is able to establish a productive infection, resulting in ultrastructural changes in cell morphology and impaired neuronal differentiation. Ultrastructural analysis of neurospheres infected with CHIKV during neuronal differentiation revealed diminished neuron dendrite formation, accumulation of viral particles associated with the plasma membrane, numerous cell vacuoles, and swollen mitochondria. Apoptotic cells were significantly increased at 72 h post-infection. Compared to Zika virus, a well-characterized neurotropic arbovirus, CHIKV infection resulted in a more discrete, albeit detectable upregulation of IL-6 levels. Finally, we found that CHIKV infection resulted in an altered profile expression, mainly downregulation, of a group of transcription factors named Hox genes. Altogether our findings highlight important features of CHIKV in the CNS, as well as the feasibility of neurospheres as robust experimental models that can support further studies for novel pharmacological interventions.
基孔肯雅病毒（CHIKV）是一种节肢动物传播病毒，最近在世界许多地区引发了大规模疫情。感染通常表现为发热性自限性疾病，其特征为关节疼痛和肌痛，不过也有严重神经表现的报道。尽管CHIKV不被认为是一种真正的嗜神经病毒，但神经元、星形胶质细胞和少突胶质细胞都易受感染。在此，我们采用3D细胞培养模型，从全反式维甲酸/脑源性神经营养因子（ATRA/BNDF）分化的人神经母细胞瘤细胞中获得神经球。我们证明CHIKV能够建立有效的感染，导致细胞形态的超微结构变化以及神经元分化受损。对神经元分化过程中感染CHIKV的神经球进行超微结构分析发现，神经元树突形成减少、与质膜相关的病毒颗粒积累、大量细胞空泡以及线粒体肿胀。感染后72小时凋亡细胞显著增加。与特征明确的嗜神经虫媒病毒寨卡病毒相比，CHIKV感染导致白细胞介素-6（IL-6）水平虽可检测到但更离散地上调。最后，我们发现CHIKV感染导致一组名为Hox基因的转录因子表达谱改变，主要是下调。总之，我们的研究结果突出了CHIKV在中枢神经系统中的重要特征，以及神经球作为强大实验模型支持新型药物干预进一步研究的可行性。
