Generation of Knockout Gene-Edited Human Intestinal Organoids.

We discuss a methodology to generate and study knockout gene-edited human intestinal organoids. We describe the generation of knockout human embryonic stem cell lines that we then differentiate into mature human intestinal organoid tissue in Matrigel using several growth factors. We also discuss a pair of assays that can be used to study the integrity of the intestinal epithelial barrier of the human intestinal organoids under inflammatory stress conditions.

Relationships between total and allergen-specific serum IgE concentrations and lung function in young adults.

BACKGROUND: Prior studies have shown relationships between serum immunoglobulin E (IgE) and asthma. OBJECTIVE: To investigate relationships between total and allergen-specific IgE concentrations and lung function in young adults. METHODS: Measurements of total IgE, allergen-specific IgE to 6 common allergens, and spirometry (forced expiratory volume in one second [FEV(1)], forced vital capacity [FVC], FEV(1)/FVC, and percent change in FEV(1) after bronchodilation) were used to calculate correlations between the logarithmically transformed IgE values and measures of lung function among participants in a birth cohort not selected for risk of allergic disease stratified by current asthma, prior asthma, or no asthma. RESULTS: The 428 participants were 51.6% female, 93% white, and 18.4 (standard deviation = 0.6) years old. Forty-eight (11.2%) had current asthma, 55 (12.9%) had a history of asthma, and 325 (75.9%) never had asthma. For males with current asthma, correlations between total IgE and FEV(1)% and FVC% were -0.51 (P = .06) and -0.70 (P = .005), respectively. For females with current asthma, the only significant correlation was between total IgE and the FEV(1)/FVC ratio (-0.55, P = .001). After excluding smokers and individuals without detectable allergen-specific IgE, the negative correlations for both males and females remained statistically significant. The correlations among males or females with prior asthma or no history of asthma were minimal and not statistically significant. The sum of the allergen-specific IgEs showed the same pattern of relationships to lung function as did total IgE. CONCLUSION: Our results show significant negative correlations that vary by gender between both total and allergen-specific IgE and measurements of lung function in young adults with current asthma.