The immediate comparison of African and Asian isolates of the power was revealed with the ZIKV from the Asian ZIKV isolate, SPH, to disturb hNP and neuronal cell function. that most likely donate to the pathological system of congenital Zika symptoms. The authors discovered that infections with Asian-lineage ZIKV isolates impaired the migration and proliferation of hNP cells, and neuron maturation. On the other hand, the African-lineage (R)-MG-132 attacks led to abrupt and extensive cell death. This work furthers the understanding of ZIKV-induced brain pathology. [33]. All measurements were compared by ANOVA and Tukey honest significant difference (HSD) test. A < 0.05. 3.2. Isolate-Specific Cell Death and Growth in hNP-Derived Neurons The differentiation process from hNP to highly enriched mature neurons requires 28 days (28 DIV), whereas nascent neurons are evident halfway through the process (14 DIV). The authors, using their well-characterized hNP to neuron differentiation process, could characterize the pathogenic effects of ZIKV infection on (R)-MG-132 populations of both immature and mature neurons. After 14 DIV in the absence of FGF2, hNP cell cultures presented a neuronal phenotype with reduced SOX1 expression and a portion of HuC/HuD+ and III-tubulin+ cells, indicative of maturing neurons (Figure 1A) [28]. The full 28 DIV of differentiation in vitro resulted in a highly homogeneous population of post-mitotic (R)-MG-132 and mature neurons characterized by microtubule-associated protein 2 (MAP2) expression (Figure 1A) [34,36]. One African Zika isolate, IbH, and one Asian isolate, SPH, were selected to evaluate the isolate-specific effects of ZIKV on 14 Rabbit Polyclonal to BRCA2 (phospho-Ser3291) DIV and 28 DIV neurons. The phenotypic differences between ZIKV lineages were again apparent when immature neurons were infected with African and Asian ZIKV. The African isolate, IbH, quickly reached peak viral production four days following infection. Peak IbH titers were followed by a marked reduction in viral production caused by virus-induced death of the cell population (Figure 2A,B). In contrast, cells infected with SPH continued to produce virions, resulting in higher viral titers peaking six or eight days post-infection. SPH-induced cell death was less apparent than with IbH, resulting in more viable immature neurons six days post-infection (Figure 2A,B). Mature neurons infected with SPH produced similar phenotypes to the nascent 14 DIV neurons (Figure 2C,D). Altogether, both isolates of the ZIKV effectively replicated in immature and mature neurons, although the Asian lineage produced higher viral titers while inducing less cell death. Open in a separate window Figure 2 ZIKV isolate-specific growth and cytotoxicity in human neurons. (A,B) Viral replication and viability of hNP-derived nascent neurons (14 DIV) and (C,D) mature neurons (28 DIV) six days post-infection. * demonstrates < 0.05. 3.3. ZIKV Infects Neural Progenitor Cells and Mature Neurons The differences observed in (R)-MG-132 cell viability between the ZIKV lineages may be influenced by the isolates abilities to initially infect the cells. A previous study found that African isolates were able to infect significantly more hNP cells than an Asian isolate [37,38]. To evaluate the ability of two prototypical Asian and African ZIKV isolates to infect hNP cells and mature neurons, the number of hNP cell or neurons containing ZIKV E protein was compared to infected Vero cells 12 h after infection. Vero cells readily produced viral proteins, and pervasive infection was observed independent of the ZIKV isolate (Figure 3A,D). hNP cells and 28 DIV mature neurons displayed lower susceptibility/permissivity relative to Vero cells. (R)-MG-132 Only a fraction of the hNP cells and 28 DIV neurons expressed ZIKV E protein, and lineage-specific susceptibility was observed in these cell lines. The high MOI infection (MOI 10) of hNP cells resulted in 13% of the IbH-infected cells expressing ZIKV E protein after 48 h, whereas only 7% of.