Analysis of the Immune Response by Standardized Whole-Blood Stimulation with Metabolism Modulation.

The immune system defends the body from infection and plays a vital role in a wide range of health conditions. Metabolism affects a series of physiological processes, including those linked to the function of human immune system. Cellular metabolism modulates immune cell activation and cytokine production. Understanding the relationship between metabolism and immune response has important implications for the development of immune-based therapeutics. However, the deployment of large-scale functional assays to investigate the metabolic regulation of immune response has been limited by the lack of standardized procedures. Here, we present a protocol for the analysis of immune response using standardized whole-blood stimulation with metabolism modulation. Diverse immune stimuli including pattern recognition receptor (PRR) ligands and microbial stimuli were incubated with fresh human whole blood. The metabolic inhibitors were used to modulate metabolic status in the immune cells. The variable immune responses after metabolic interventions were evaluated. We described in detail the main steps involved in the whole-blood stimulation and cytokines quantification, namely, collection and treatment of whole blood, preparation of samples and controls, cytokines detection, and stimulation with metabolic interventions. The metabolic inhibitors for anabolic pathways and catabolic pathways exert selective effects on the production of cytokines from immune cells. In addition to a robust and accurate assessment of immune response in cohort studies, the standardized whole-blood stimulation with metabolic regulation might provide new insights for modulating immunity. Supplementary Information: The online version contains supplementary material available at 10.1007/s43657-023-00114-0.

Phonological Development in 3-6-Year-Old Mandarin-Speaking Children with Autism, Developmental Delays, and Typical Development.

Research on the phonological development of children with autism spectrum disorder (ASD) has not yet reached consistent conclusions, and systematic studies from different language groups are needed. This study aimed to systematically investigate the characteristics of phonological development in 3-6 year-old Mandarin-speaking children with ASD. We analyzed 10 min speech samples from 21 children with ASD, 18 development level-matched children with developmental disorders (DD), and 15 chronological age-matched typically developing (TD) children during semi-structured parent-child free play based on Mandarin phonological features. The children with ASD had a significantly smaller inventory than those with TD on the initial and final inventories. The children with ASD had only a significantly smaller initial inventory than those with DD in Phases 2 and 4. Compared with TD children, children with ASD used a higher proportion of V1 and V1V2C and a smaller proportion of V1V2V3, CV1C, and CV1V2C. No significant differences existed between ASD and DD children in the proportion of any syllable structure, but V1V2V3, CV1, and CV1V2C numbers were significantly fewer than in DD children. Children with ASD were significantly greater than children with TD in the diversity of V1V2, CV1, and overall syllables. ASD children had significantly fewer different types of syllables in both V1V2C and CV1 than did DD children and significantly greater diversity in CV1 and overall syllables than did DD children. These preliminary data suggest that the gap between TD and ASD children's language abilities increased with age, and this gap was reflected in initial, final, and syllable complexity and diversity. Children with DD and ASD showed similar language abilities, and children with DD showed detailed differences from those with ASD regarding initial, syllable complexity and diversity.