SeV C Protein Plays a Role in Restricting Macrophage Phagocytosis by Limiting the Generation of Intracellular Double-Stranded RNA.

Macrophages play a central role in the innate immune response to respiratory viral infections through pro-inflammatory factor secretion and phagocytosis. However, as a countermeasure, viral pathogens have evolved virulence factors to antagonize macrophage function. In our recent in vitro analyses of murine macrophage cell lines, Sendai virus (SeV) accessory protein C inhibited the secretion of pro-inflammatory factors, and C gene-knockout SeV (SeVΔC) caused drastic morphological changes in RAW264.7 macrophages, similar to those observed after stimulation with Lipid A, a well-known activator of actin-rich membrane ruffle formation and phagocytosis. Hence, we sought to determine whether the C protein limits phagocytosis in SeV-infected macrophages through the suppression of membrane ruffling. Phagocytosis assays indicated an upregulation of phagocytosis in both SeVΔC-infected and Lipid A-stimulated macrophages, but not in SeV WT-infected cells. Further, the observed membrane ruffling was associated with phagocytosis. RIG-I is essential for Lipid A-induced phagocytosis; its deficiency inhibited SeVΔC-stimulated phagocytosis and ruffling, confirming the essential role of RIG-I. Moreover, treatment with interferon (IFN)-ß stimulation and neutralizing antibodies against IFN-ß suggested that SeVΔC-induced phagocytosis and ruffling occurred in an IFN-ß-independent manner. A newly isolated SeVΔC strain that does not generate dsRNA further highlighted the importance of dsRNA in the induction of phagocytosis and ruffling. Taken together, the current results suggest that SeV C protein might limit phagocytosis-associated membrane ruffling in an RIG-I-mediated but IFN-independent manner via limiting the generation of intracellular dsRNA.

Sendai virus C protein affects macrophage function, which plays a critical role in modulating disease severity during Sendai virus infection in mice.

Sendai virus (SeV) accessory protein C limits the generation of double-stranded RNAs, defective interfering RNAs, or both, during viral transcription and replication, thereby limiting interferon-ß production. Our recent in vitro analyses on murine macrophage cell lines demonstrated that this protein also contributes to restricting macrophage function, including the production of nitric oxide (NO) and inflammatory cytokines in addition to interferon-ß, in infected macrophages. This study showed that depletion of airway macrophages by clodronate-loaded liposomes led to the development of severe viral pneumonia in recombinant C gene-knockout SeV (SeV∆C)-infected mice, but did not modulate disease severity in wild-type SeV-infected mice. Furthermore, the severe disease observed in macrophage-depleted, SeV∆C-infected mice was associated with exacerbated virus replication in the lungs, leading to severe airway inflammation and pulmonary edema, indicating lung injury. These results suggested that the antimacrophage activity of SeV C protein might play a critical role in modulating lung injury and associated diseases caused by SeV.

Timing of phase-amplitude coupling is essential for neuronal and functional maturation of audiovisual integration in adolescents.

OBJECTIVE: The ability to integrate audiovisual information matures late in adolescents, but its neuronal mechanism is still unknown. Recent studies showed that phase-amplitude coupling (PAC) of neuronal oscillations, which is defined as the modulation of high-frequency amplitude by low-frequency phase, is associated with audiovisual integration in adults. Thus, we investigated how PAC develops in adolescents and whether it is related to the functional maturation of audiovisual integration. In particular, we focused on the timing of PAC (or the coupling phase), which is defined as the low-frequency phase with maximum high-frequency amplitude. METHODS: Using magnetoencephalography (MEG) on 15 adults and 14 adolescents while they performed an audiovisual speech integration task, we examined PAC in association cortexes with a trial-by-trial analysis. RESULTS: Whereas delta-beta coupling was consistently observed in both adults and adolescents, we found that the timing of delta-beta PAC was delayed by 20-40 milliseconds in adolescents compared with adults. In addition, a logistic regression analysis revealed that the task performance improves as the timing of delta-beta PAC in the right temporal pole (TP) got closer to the trough position (180 degrees). CONCLUSION: These results suggest that the timing of PAC is essential for binding audiovisual information and underlies the developmental process in adolescents.

Uniqueness of action monitoring in children with autism spectrum disorder: Response types and temporal aspects.

BACKGROUND: Action monitoring, the process for evaluating the appropriateness of one's own actions, is reported to be atypical in individuals with autism spectrum disorder (ASD). METHOD: We examined the characteristics of action monitoring in 11 children with ASD and 12 children with typical development (TD), analyzing stimulus-locked and response-locked event-related potential components (i.e., N2; error-related negativity, ERN; and error positivity, Pe) related to execution of a flanker task. RESULTS: We found a smaller N2 amplitude in children with ASD than in those with TD. Children with ASD also had a larger amplitude of ERN for partial error responses (electromyographic activity corresponding to the inappropriate hand side before response execution) than did children with TD. Additionally, the ERN amplitude for the partial error response was correlated with the Autistic Mannerisms of the Social Responsiveness Scale. There were no significant differences in Pe amplitudes between children with ASD and those with TD. CONCLUSION: The results suggest that action monitoring in children with ASD is significantly different both before and after response execution. We hypothesized that the detail-focused processing style of ASD reduces the demands of action monitoring before response execution; however, autistic mannerisms evoke excessive concern regarding trivial mistakes after response execution.

Facial identity recognition in children with autism spectrum disorders revealed by P300 analysis: a preliminary study.

BACKGROUND: To reveal the neural substrate of communication difficulties in children with autism spectrum disorders (ASDs), we investigated the P300 component of event-related potentials (ERPs) as represented by the average of electroencephalography findings time-locked to events and behavior. Because the P300 amplitude influences attentional resource allocation during discrimination, the component elicited during perception of known and unknown faces should indicate familiarity processing. METHODS AND RESULTS: Nine typically developing children (TD) and nine children with ASDs participated in this study (Experiment 1). The P300 amplitude in TD children was significantly larger during familiar face perception than during unfamiliar face perception (p<0.01). However, there was no evidence of familiarity effect in children with ASDs. In three children with ASDs, we also assessed the P300 amplitude during perception of a therapist's face one month before (baseline), a few days before and after social skills training (SST) sessions (Experiment 2). To evaluate the effect of familiarity on facial identity processing, we analyzed the therapist/unknown ratio of P300 amplitudes related to the face discrimination task as an index. The ratio was larger after SST sessions than before, but there was no difference in the ratio between baseline and before SST assessments. CONCLUSION: The P300 might be influenced by attentional resource allocation depending on the stage of learning face identification in children with ASDs. We speculate that this approach to evaluating brain responses during facial identity recognition could be used as a tool to clarify children's communication difficulties.

[Effectiveness of social skills training for children with developmental disorders: behavioral analysis using a two-dimensional motion capture system].

The current study sought to develop a new behavioral analysis methods to evaluate the effects of social skills training (SST). SST is known to be an effective method to improve the social skills of children with behavioral problems. However, current evaluation methods involve behavioral rating scales that are heavily dependent on evaluators' particular experiences they have had. To quantitatively examine the behavioral effects of SST, we examined subjects' head-movements related to social behavior, using a two-dimensional motion capture system (Kissei Comtec, Japan). Four children (three male, one female, 7-8 years of age) with pervasive developmental disorder (PDD) or attention deficit/hyperactivity disorder (AD/HD) participated in 16 sessions of SST. Before and after SST, head-coordinates on a two-dimensional plane were calculated using their behavior during a pair task, measured by four digital cameras. After SST, the number of communication behaviors was increased compared to before SST. In addition, children looked longer at another child within 30 degrees of the central visual field. Time-series analysis of the visual field during the detection of another child revealed significant auto-correlation from about -1.12 second. before to the beginning of communication behavior (p<0.05). The results suggested that our method can provide a quantitative index of characteristics related to skilled social behaviors. We conclude that a two-dimensional motion capture system would be useful for visualization of the interventional effects of SST, which would supplement assessments by the conventional observational strategies.